WorldWideScience

Sample records for assisted laser desorption

  1. Investigations into ultraviolet matrix-assisted laser desorption

    Energy Technology Data Exchange (ETDEWEB)

    Heise, T.W.

    1993-07-01

    Matrix-assisted laser desorption (MALD) is a technique for converting large biomolecules into gas phase ions. Some characteristics of the commonly used uv matrices are determined. Solubilities in methanol range from 0.1 to 0.5 M. Solid phase absorption spectra are found to be similar to solution, but slightly red-shifted. Acoustic and quartz crystal microbalance signals are investigated as possible means of uv-MALD quantitation. Evidence for the existence of desorption thresholds is presented. Threshold values are determined to be in the range of 2 to 3 MW/cm{sup 2}. A transient imaging technique based on laser-excited fluorescence for monitoring MALD plumes is described. Sensitivity is well within the levels required for studying matrix-assisted laser desorption, where analyte concentrations are significantly lower than those in conventional laser desorption. Results showing the effect of film morphology, particularly film thickness, on plume dynamics are presented. In particular, MALD plumes from thicker films tend to exhibit higher axial velocities. Fluorescent labeling of protein and of DNA is used to allow imaging of their uv-MALD generated plumes. Integrated concentrations are available with respect to time, making it possible to assess the rate of fragmentation. The spatial and temporal distributions are important for the design of secondary ionization schemes to enhance ion yields and for the optimization of ion collection in time-of-flight MS instruments to maximize resolution. Such information could also provide insight into whether ionization is closely associated with the desorption step or whether it is a result of subsequent collisions with the matrix gas (e.g., proton transfer). Although the present study involves plumes in a normal atmosphere, adaptation to measurements in vacuum (e.g., inside a mass spectrometer) should be straightforward.

  2. Surface-assisted laser desorption ionization mass spectrometry techniques for application in forensics.

    Science.gov (United States)

    Guinan, Taryn; Kirkbride, Paul; Pigou, Paul E; Ronci, Maurizio; Kobus, Hilton; Voelcker, Nicolas H

    2015-01-01

    Matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS) is an excellent analytical technique for the rapid and sensitive analysis of macromolecules (>700 Da), such as peptides, proteins, nucleic acids, and synthetic polymers. However, the detection of smaller organic molecules with masses below 700 Da using MALDI-MS is challenging due to the appearance of matrix adducts and matrix fragment peaks in the same spectral range. Recently, nanostructured substrates have been developed that facilitate matrix-free laser desorption ionization (LDI), contributing to an emerging analytical paradigm referred to as surface-assisted laser desorption ionization (SALDI) MS. Since SALDI enables the detection of small organic molecules, it is rapidly growing in popularity, including in the field of forensics. At the same time, SALDI also holds significant potential as a high throughput analytical tool in roadside, work place and athlete drug testing. In this review, we discuss recent advances in SALDI techniques such as desorption ionization on porous silicon (DIOS), nano-initiator mass spectrometry (NIMS) and nano assisted laser desorption ionization (NALDI™) and compare their strengths and weaknesses with particular focus on forensic applications. These include the detection of illicit drug molecules and their metabolites in biological matrices and small molecule detection from forensic samples including banknotes and fingerprints. Finally, the review highlights recent advances in mass spectrometry imaging (MSI) using SALDI techniques.

  3. Quantitative analysis of biopolymers by matrix-assisted laser desorption

    Energy Technology Data Exchange (ETDEWEB)

    Tang, K.; Allman, S.L.; Jones, R.B.; Chen, C.H. (Oak Ridge National Lab., TN (United States))

    1993-08-01

    During the past few years, major efforts have been made to use mass spectrometry to measure biopolymers because of the great potential benefit to biological and medical research. Although the theoretical details of laser desorption and ionization mechanisms of MALDI are not yet fully understood, several models have been presented to explain the production of large biopolymer ions. In brief, it is very difficult to obtain reliable measurements of the absolute quantity of analytes by MALDI. If MALDI is going to become a routine analytical tool, it is obvious that quantitative measurement capability must be pursued. Oligonucleotides and protein samples used in this work were purchased from commercial sources. Nicotinic acid was used as matrix for both types of biopolymers. From this experiment, it is seen that it is difficult to obtain absolute quantitative measurements of biopolymers using MALDI. However, internal calibration with molecules having similar chemical properties can be used to resolve these difficulties. Chemical reactions between biopolymers must be avoided to prevent the destruction of the analyte materials. 10 refs., 8 figs.

  4. Characterisation of bacteria by matrix-assisted laser desorption/ionisation and electrospray mass spectrometry

    NARCIS (Netherlands)

    Baar, B.L.M. van

    2000-01-01

    Chemical analysis for the characterisation of micro-organisms is rapidly evolving, after the recent advent of new ionisation methods in mass spectrometry (MS): electrospray (ES) and matrix-assisted laser desorption/ionisation (MALDI). These methods allow quick characterisation of micro-organisms, ei

  5. Derivatization of small biomolecules for optimized matrix-assisted laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Tholey, Andreas; Wittmann, Christoph; Kang, Min-Jung; Bungert, Ditte; Hollemeyer, Klaus; Heinzle, Elmar

    2002-09-01

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) is a powerful tool for the measurement of low molecular mass compounds of biological interest. The limitations for this method are the volatility of many analytes, possible interference with matrix signals or bad ionization or desorption behavior of the compounds. We investigated the application of well-known and straightforward one-pot derivatization procedures to circumvent these problems. The derivatizations tested allow the measurement and the labeling of alcohols, aldehydes and ketones, carboxylic acids, alpha-ketocarboxylic acids and amines.

  6. Unusual Fragmentation of Peptide and Protein in Matrix-assisted Laser Desorption/Ionization Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    Mitsuo Takayama

    2001-01-01

    Unusual amine - bond fragmentation on the peptide/protein backbone has been reported using matrix - assisted laser desorption/ionization time - of- flight mass spectrometry (MALDI - TOFMS)The amine - bond cleavage occurred without metastable decay, while the peptide - bond cleavage occurred with metastable decay of peptide ions in a drift region of TOF mass analyzer. It was presumed that the amine - bond cleavage occurred as a non - ergodic process independent of the ionization under MALDI conditions.

  7. Electrospray ionization and matrix assisted laser desorption/ionization mass spectrometry: powerful analytical tools in recombinant protein chemistry

    DEFF Research Database (Denmark)

    Andersen, Jens S.; Svensson, B; Roepstorff, P

    1996-01-01

    Electrospray ionization and matrix assisted laser desorption/ionization are effective ionization methods for mass spectrometry of biomolecules. Here we describe the capabilities of these methods for peptide and protein characterization in biotechnology. An integrated analytical strategy is presen......Electrospray ionization and matrix assisted laser desorption/ionization are effective ionization methods for mass spectrometry of biomolecules. Here we describe the capabilities of these methods for peptide and protein characterization in biotechnology. An integrated analytical strategy...

  8. Pyroelectricity Assisted Infrared-Laser Desorption Ionization (PAI-LDI) for Atmospheric Pressure Mass Spectrometry

    Science.gov (United States)

    Li, Yanyan; Ma, Xiaoxiao; Wei, Zhenwei; Gong, Xiaoyun; Yang, Chengdui; Zhang, Sichun; Zhang, Xinrong

    2015-08-01

    A new atmospheric pressure ionization method termed pyroelectricity-assisted infrared laser desorption ionization (PAI-LDI) was developed in this study. The pyroelectric material served as both sample target plate and enhancing ionization substrate, and an IR laser with wavelength of 1064 nm was employed to realize direct desorption and ionization of the analytes. The mass spectra of various compounds obtained on pyroelectric material were compared with those of other substrates. For the five standard substances tested in this work, LiNbO3 substrate produced the highest ion yield and the signal intensity was about 10 times higher than that when copper was used as substrate. For 1-adamantylamine, as low as 20 pg (132.2 fmol) was successfully detected. The active ingredient in (Compound Paracetamol and 1-Adamantylamine Hydrochloride Capsules), 1-adamantylamine, can be sensitively detected at an amount as low as 150 pg, when the medicine stock solution was diluted with urine. Monosaccharide and oligosaccharides in Allium Cepa L. juice was also successfully identified with PAI-LDI. The method did not require matrix-assisted external high voltage or other extra facility-assisted set-ups for desorption/ionization. This study suggested exciting application prospect of pyroelectric materials in matrix- and electricity-free atmospheric pressure mass spectrometry research.

  9. Measurement of Chiral Recognition Properties of Crown Ethers Using Matrix Assisted Laser Desorption Ionization Mass Spectrometry

    OpenAIRE

    SAWADA, Masami; Harada, Manabu; TAKAI, Yoshio; NAKANO, Kazurou; Kuroda, Masao; ARAKAWA, Ryuichi; 荒川, 隆一

    2000-01-01

    Hydrogen bonding host-guest complex ions between chiral crown ethers and chiral amino acid ester salts, detected by matrix assisted laser desorption ionization (MALDI) mass spectrometry (MS) with a DHBA or MSA matrix, were studied on the view point of chiral recognition properties of the chiral crown hosts. The chiral recognition property (IR/IS-dn value≅1.0) obtained by the present MALDI-MS is sharply different from the IR/IS-dn value obtained by FAB-MS or ESI-MS (≠1.0) in the same host-gues...

  10. Cosmetic Analysis Using Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI-MSI

    Directory of Open Access Journals (Sweden)

    Rodrigo Ramos Catharino

    2013-03-01

    Full Text Available A new “omic” platform—Cosmetomics—that proves to be extremely simple and effective in terms of sample preparation and readiness for data acquisition/interpretation is presented. This novel approach employing Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI-MSI for cosmetic analysis has proven to readily identify and quantify compounds of interest. It also allows full control of all the production phases, as well as of the final product, by integration of both analytical and statistical data. This work has focused on products of daily use, namely nail polish, lipsticks and eyeliners of multiple brands sold in the worldwide market.

  11. Matrix-assisted laser desorption mass spectrometry of gas-phase peptide-metal complexes

    Science.gov (United States)

    Hortal, Ana R.; Hurtado, Paola; Martínez-Haya, Bruno

    2008-12-01

    Cation attachment to a model peptide has been investigated in matrix-assisted laser desorption experiments. Angiotensin I (Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu) is chosen as a system for study, and Cu2+ and K+ salts are used as cationizing agents. Three fundamentally different types of samples are investigated: (1) a crystalline sample of Ang I, metal salt and MALDI matrix, prepared with the conventional dried droplet method; (2) a solvent-free fine powder mixture of the same three compounds, and (3) a solution of the angiotensin and the metal salt in an ionic liquid matrix (a molten organic salt that acts as a MALDI active solvent). Effective protonation and cationization of the peptide are achieved with the three methods. The transition metal systematically provides more efficient cationization than the alkali metal. At sufficiently high concentration of the salt, the attachment of up to four copper cations to the angiotensin is observed in the MALDI spectrum. In contrast, only one K+ cation is efficiently bound to the peptide. For a given salt concentration, the highest degree of cationization is obtained in the laser desorption from the ionic liquid matrix. This is attributed to the efficient transfer of free metal cations to the desorption plume, where the complexation takes place.

  12. Revisiting the quantitative features of surface-assisted laser desorption/ionization mass spectrometric analysis.

    Science.gov (United States)

    Wu, Ching-Yi; Lee, Kai-Chieh; Kuo, Yen-Ling; Chen, Yu-Chie

    2016-10-28

    Surface-assisted laser desorption/ionization (SALDI) coupled with mass spectrometry (MS) is frequently used to analyse small organics owing to its clean background. Inorganic materials can be used as energy absorbers and the transfer medium to facilitate the desorption/ionization of analytes; thus, they are used as SALDI-assisting materials. Many studies have demonstrated the usefulness of SALDI-MS in quantitative analysis of small organics. However, some characteristics occurring in SALDI-MS require certain attention to ensure the reliability of the quantitative analysis results. The appearance of a coffee-ring effect in SALDI sample preparation is the primary factor that can affect quantitative SALDI-MS analysis results. However, to the best of our knowledge, there are no reports relating to quantitative SALDI-MS analysis that discuss or consider this effect. In this study, the coffee-ring effect is discussed using nanoparticles and nanostructured substrates as SALDI-assisting materials to show how this effect influences SALDI-MS analysis results. Potential solutions for overcoming the existing problems are also suggested.This article is part of the themed issue 'Quantitative mass spectrometry'.

  13. Ion intensity and thermal proton transfer in ultraviolet matrix-assisted laser desorption/ionization.

    Science.gov (United States)

    Lu, I-Chung; Lee, Chuping; Chen, Hui-Yuan; Lin, Hou-Yu; Hung, Sheng-Wei; Dyakov, Yuri A; Hsu, Kuo-Tung; Liao, Chih-Yu; Lee, Yin-Yu; Tseng, Chien-Ming; Lee, Yuan-Tseh; Ni, Chi-Kung

    2014-04-17

    The ionization mechanism of ultraviolet matrix-assisted laser desorption/ionization (UV-MALDI) was investigated by measuring the total cation intensity (not including sodiated and potasiated ions) as a function of analyte concentration (arginine, histidine, and glycine) in a matrix of 2,4,6-trihydroxyacetophenone (THAP). The total ion intensity increased up to 55 times near the laser fluence threshold as the arginine concentration increased from 0% to 1%. The increases were small for histidine, and a minimal increase occurred for glycine. Time-resolved fluorescence intensity was employed to investigate how analytes affected the energy pooling of the matrix. No detectable energy pooling was observed for pure THAP and THAP/analyte mixtures. The results can be described by using a thermal proton transfer model, which suggested that thermally induced proton transfer is crucial in the primary ion generation in UV-MALDI.

  14. Quantum dots assisted laser desorption/ionization mass spectrometric detection of carbohydrates: qualitative and quantitative analysis.

    Science.gov (United States)

    Bibi, Aisha; Ju, Huangxian

    2016-04-01

    A quantum dots (QDs) assisted laser desorption/ionization mass spectrometric (QDA-LDI-MS) strategy was proposed for qualitative and quantitative analysis of a series of carbohydrates. The adsorption of carbohydrates on the modified surface of different QDs as the matrices depended mainly on the formation of hydrogen bonding, which led to higher MS intensity than those with conventional organic matrix. The effects of QDs concentration and sample preparation method were explored for improving the selective ionization process and the detection sensitivity. The proposed approach offered a new dimension to the application of QDs as matrices for MALDI-MS research of carbohydrates. It could be used for quantitative measurement of glucose concentration in human serum with good performance. The QDs served as a matrix showed the advantages of low background, higher sensitivity, convenient sample preparation and excellent stability under vacuum. The QDs assisted LDI-MS approach has promising application to the analysis of carbohydrates in complex biological samples.

  15. Manganese oxide nanoparticle-assisted laser desorption/ionization mass spectrometry for medical applications

    Directory of Open Access Journals (Sweden)

    Shu Taira, Kenji Kitajima, Hikaru Katayanagi, Eiichiro Ichiishi and Yuko Ichiyanagi

    2009-01-01

    Full Text Available We prepared and characterized manganese oxide magnetic nanoparticles (d =5.6 nm and developed nanoparticle-assited laser desorption/ionization (nano-PALDI mass spectrometry. The nanoparticles had MnO2 and Mn2O3 cores conjugated with hydroxyl and amino groups, and showed paramagnetism at room temperature. The nanoparticles worked as an ionization assisting reagent in mass spectroscopy. The mass spectra showed no background in the low m/z. The nanoparticles could ionize samples of peptide, drug and proteins (approx. 5000 Da without using matrix, i.e., 2,5-dihydroxybenzoic acid (DHB, 4-hydroxy-α-cinnamic acid (CHCA and liquid matrix, as conventional ionization assisting reagents. Post source decay spectra by nano-PALDI mass spectrometry will yield information of the chemical structure of analytes.

  16. Beer fingerprinting by Matrix-Assisted Laser Desorption-Ionisation-Time of Flight Mass Spectrometry.

    Science.gov (United States)

    Šedo, Ondrej; Márová, Ivana; Zdráhal, Zbyněk

    2012-11-15

    A method allowing parallel fingerprinting of proteins and maltooligosaccharides directly from untreated beer samples is presented. These two classes of compounds were detected by Matrix-Assisted Laser Desorption-Ionisation-Time of Flight-Mass Spectrometry (MALDI-TOF-MS) analysis of beer mixed with 2,5-dihydroxybenzoic acid solution. The maltooligosaccharide profiles acquired from the MALDI sample spot center were not found characteristic for beers of different source and technology. On the other hand, according to profiles containing protein signals acquired from crystals formed on the border of the MALDI sample spot, we were able to distinguish beer samples of the same brand produced by different breweries. The discriminatory abilities of the method were further examined on a set of 17 lager beers, where the fingerprints containing protein signals enabled resolution of majority of examined brands. We propose MALDI-TOF-MS profiling as a rapid tool for beer brewing technology process monitoring, quality control, and determination of beer authenticity.

  17. Direct Surface Analysis of Fungal Species by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Valentine, Nancy B.(BATTELLE (PACIFIC NW LAB)); Wahl, Jon H.(BATTELLE (PACIFIC NW LAB)); Kingsley, Mark T.(BATTELLE (PACIFIC NW LAB)); Wahl, Karen L.(BATTELLE (PACIFIC NW LAB))

    2001-12-01

    Intact spores and/or hyphae of Aspergillus niger, Rhizopus oryzae, Trichoderma reesei and Phanerochaete chrysosporium are analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). This study investigates various methods of sample preparation and matrices to determine optimum collection and analysis criteria for fungal analysis by MALDI-MS. Fungi are applied to the MALDI sample target as untreated, sonicated, acid/heat treated, or blotted directly from the fungal culture with double-stick tape. Ferulic acid or sinapinic acid matrix solution is layered over the dried samples and analyzed by MALDI-MS. Statistical analysis of the data show that simply using double stick tape to collect and transfer to a MALDI sample plate typically worked as well as the other preparation methods, but requires the least sample handling.

  18. omniSpect: an open MATLAB-based tool for visualization and analysis of matrix-assisted laser desorption/ionization and desorption electrospray ionization mass spectrometry images.

    Science.gov (United States)

    Parry, R Mitchell; Galhena, Asiri S; Gamage, Chaminda M; Bennett, Rachel V; Wang, May D; Fernández, Facundo M

    2013-04-01

    We present omniSpect, an open source web- and MATLAB-based software tool for both desorption electrospray ionization (DESI) and matrix-assisted laser desorption ionization (MALDI) mass spectrometry imaging (MSI) that performs computationally intensive functions on a remote server. These functions include converting data from a variety of file formats into a common format easily manipulated in MATLAB, transforming time-series mass spectra into mass spectrometry images based on a probe spatial raster path, and multivariate analysis. OmniSpect provides an extensible suite of tools to meet the computational requirements needed for visualizing open and proprietary format MSI data.

  19. Basic matrices in the analysis of non-covalent complexes by matrix-assisted laser desorption/ionization mass spectrometry

    NARCIS (Netherlands)

    Jespersen, S.; Niessen, W.M.A.; Tjaden, U.R.; Greef, J. van der

    1998-01-01

    A number of potential matrix candidates were investigated with regard to the importance of the pH in the matrix-assisted laser desorption/ionization mass spectrometry (MALDI/MS) analysis of non-covalently bound protein complexes. The matrices examined were 2,5-dihydroxybenzoic acid (DHB), 4-hydroxy-

  20. Development of matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) for plant metabolite analysis

    Energy Technology Data Exchange (ETDEWEB)

    Korte, Andrew R [Iowa State Univ., Ames, IA (United States)

    2014-12-01

    This thesis presents efforts to improve the methodology of matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) as a method for analysis of metabolites from plant tissue samples. The first chapter consists of a general introduction to the technique of MALDI-MSI, and the sixth and final chapter provides a brief summary and an outlook on future work.

  1. Application of matrix-assisted laser desorption/ionization to on-line aerosol time-of-flight mass spectrometry

    NARCIS (Netherlands)

    Stowers, M.A.; Wuijckhuijse, A.L. van; Marijnissen, J.C.M.; Scarlett, B.; Baar, B.L.M. van; Kientz, Ch.E.

    2000-01-01

    Matrix-assisted laser desorption/ionization (MALDI) mass spectra were obtained from single biological aerosol particles using an aerosol time-of- flight mass spectrometer (ATOFMS). The inlet to the ATOFMS was coupled with an evaporation/condensation flow cell that allowed the aerosol to be coated wi

  2. Performance of matrix-assisted laser desorption-time of flight mass spectrometry for identification of clinical yeast isolates

    DEFF Research Database (Denmark)

    Rosenvinge, Flemming S; Dzajic, Esad; Knudsen, Elisa;

    2013-01-01

    Accurate and fast yeast identification is important when treating patients with invasive fungal disease as susceptibility to antifungal agents is highly species related. Matrix-assisted laser desorption-time of flight mass spectrometry (MALDI-TOF-MS) provides a powerful tool with a clear potential...

  3. A novel sample preparation method of matrix-assisted laser desorption/ionization time of flight mass spectrometry for polystyrene

    Institute of Scientific and Technical Information of China (English)

    Shu Zhang; Zhen Wen Zhao; Lei Xiong; Bin Xin; Wei Hua Hu; Shao Xiang Xiong

    2007-01-01

    A novel sample preparation method of matrix-assisted laser desorption/ionization mass spectrometry for polystyrene was reported.Compared to the conventional dried-droplet method, the efficiency of ionization and signal intensity of mass spectra were improved.The mechanism was also analyzed.

  4. Thermal proton transfer reactions in ultraviolet matrix-assisted laser desorption/ionization.

    Science.gov (United States)

    Chu, Kuan Yu; Lee, Sheng; Tsai, Ming-Tsang; Lu, I-Chung; Dyakov, Yuri A; Lai, Yin Hung; Lee, Yuan-Tseh; Ni, Chi-Kung

    2014-03-01

    One of the reasons that thermally induced reactions are not considered a crucial mechanism in ultraviolet matrix-assisted laser desorption ionization (UV-MALDI) is the low ion-to-neutral ratios. Large ion-to-neutral ratios (10(-4)) have been used to justify the unimportance of thermally induced reactions in UV-MALDI. Recent experimental measurements have shown that the upper limit of the total ion-to-neutral ratio is approximately 10(-7) at a high laser fluence and less than 10(-7) at a low laser fluence. Therefore, reexamining the possible contributions of thermally induced reactions in MALDI may be worthwhile. In this study, the concept of polar fluid was employed to explain the generation of primary ions in MALDI. A simple model, namely thermal proton transfer, was used to estimate the ion-to-neutral ratios in MALDI. We demonstrated that the theoretical calculations of ion-to-neutral ratios exhibit the same trend and similar orders of magnitude compared with those of experimental measurements. Although thermal proton transfer may not generate all of the ions observed in MALDI, the calculations demonstrated that thermally induced reactions play a crucial role in UV-MALDI.

  5. Formation of Metal-Related Ions in Matrix-Assisted Laser Desorption Ionization

    Science.gov (United States)

    Lee, Chuping; Lu, I.-Chung; Hsu, Hsu Chen; Lin, Hou-Yu; Liang, Sheng-Ping; Lee, Yuan-Tseh; Ni, Chi-Kung

    2016-09-01

    In a study of the metal-related ion generation mechanism in matrix-assisted laser desorption ionization (MALDI), crystals of matrix used in MALDI were grown from matrix- and salt-containing solutions. The intensities of metal ion and metal adducts of the matrix ion obtained from unwashed crystals were higher than those from crystals washed with deionized water, indicating that metal ions and metal adducts of the matrix ions are mainly generated from the surface of crystals. The contributions of preformed metal ions and metal adducts of the matrix ions inside the matrix crystals were minor. Metal adducts of the matrix and analyte ion intensities generated from a mixture of dried matrix, salt, and analyte powders were similar to or higher than those generated from the powder of dried droplet crystals, indicating that the contributions of the preformed metal adducts of the matrix and analyte ions were insignificant. Correlation between metal-related ion intensity fluctuation and protonated ion intensity fluctuation was observed, indicating that the generation mechanism of the metal-related ions is similar to that of the protonated ions. Because the thermally induced proton transfer model effectively describes the generation of the protonated ions, we suggest that metal-related ions are mainly generated from the salt dissolution in the matrix melted by the laser.

  6. Bioaerosol detection by aerosol TOF-mass spectrometry: Application of matrix assisted laser desorption/ionisation

    NARCIS (Netherlands)

    Wuijckhuijse, A.L. van; Stowers, M.A.; Kientz, Ch.E.; Marijnissen, J.C.M.; Scarlett, B.

    2000-01-01

    In previous publications the use of an aerosol time of flight mass spectrometer was reported for the on-line measurements of aerosols (Weiss 1997, Kievit 1995). The apparatus is capable of measuring the size as well as the chemical composition, by the use of Laser Desorption/Ionisation (LDI), of an

  7. Study of ionization process of matrix molecules in matrix-assisted laser desorption ionization

    Energy Technology Data Exchange (ETDEWEB)

    Murakami, Kazumasa; Sato, Asami; Hashimoto, Kenro; Fujino, Tatsuya, E-mail: fujino@tmu.ac.jp

    2013-06-20

    Highlights: ► Proton transfer and adduction reaction of matrix in MALDI were studied. ► Hydroxyl group forming intramolecular hydrogen bond was related to the ionization. ► Intramolecular proton transfer in the electronic excited state was the initial step. ► Non-volatile analytes stabilized protonated matrix in the ground state. ► A possible mechanism, “analyte support mechanism”, has been proposed. - Abstract: Proton transfer and adduction reaction of matrix molecules in matrix-assisted laser desorption ionization were studied. By using 2,4,6-trihydroxyacetophenone (THAP), 2,5-dihydroxybenzoic acid (DHBA), and their related compounds in which the position of a hydroxyl group is different, it was clarified that a hydroxyl group forming an intramolecular hydrogen bond is related to the ionization of matrix molecules. Intramolecular proton transfer in the electronic excited state of the matrix and subsequent proton adduction from a surrounding solvent to the charge-separated matrix are the initial steps for the ionization of matrix molecules. Nanosecond pump–probe NIR–UV mass spectrometry confirmed that the existence of analyte molecules having large dipole moment in their structures is necessary for the stabilization of [matrix + H]{sup +} in the electronic ground state.

  8. Black phosphorus-assisted laser desorption ionization mass spectrometry for the determination of low-molecular-weight compounds in biofluids.

    Science.gov (United States)

    He, Xiao-Mei; Ding, Jun; Yu, Lei; Hussain, Dilshad; Feng, Yu-Qi

    2016-09-01

    Quantitative analysis of small molecules by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been a challenging task due to matrix-derived interferences in low m/z region and poor reproducibility of MS signal response. In this study, we developed an approach by applying black phosphorus (BP) as a matrix-assisted laser desorption ionization (MALDI) matrix for the quantitative analysis of small molecules for the first time. Black phosphorus-assisted laser desorption/ionization mass spectrometry (BP/ALDI-MS) showed clear background and exhibited superior detection sensitivity toward quaternary ammonium compounds compared to carbon-based materials. By combining stable isotope labeling (SIL) strategy with BP/ALDI-MS (SIL-BP/ALDI-MS), a variety of analytes labeled with quaternary ammonium group were sensitively detected. Moreover, the isotope-labeled forms of analytes also served as internal standards, which broadened the analyte coverage of BP/ALDI-MS and improved the reproducibility of MS signals. Based on these advantages, a reliable method for quantitative analysis of aldehydes from complex biological samples (saliva, urine, and serum) was successfully established. Good linearities were obtained for five aldehydes in the range of 0.1-20.0 μM with correlation coefficients (R (2)) larger than 0.9928. The LODs were found to be 20 to 100 nM. Reproducibility of the method was obtained with intra-day and inter-day relative standard deviations (RSDs) less than 10.4 %, and the recoveries in saliva samples ranged from 91.4 to 117.1 %. Taken together, the proposed SIL-BP/ALDI-MS strategy has proved to be a reliable tool for quantitative analysis of aldehydes from complex samples. Graphical Abstract An approach for the determination of small molecules was developed by using black phosphorus (BP) as a matrix-assisted laser desorption ionization (MALDI) matrix.

  9. Overview literature on matrix assisted laser desorption ionization mass spectroscopy (MALDI MS): basics and its applications in characterizing polymeric materials

    Indian Academy of Sciences (India)

    R N Jagtap; A H Ambre

    2005-10-01

    Matrix assisted laser desorption ionization mass spectroscopy (MALDI MS) is a technique which allows the measurement of molecular mass > 200,000 Daltons by ionization and vapourization without degradation. This technique is useful for the mass analysis of synthetic polymers, which have very low volatility. The basic principles of and its applications for polymer characterization have been discussed in this paper. In addition, the possibilities of combining MALDI MS with chromatographic and other analytical techniques have also been discussed.

  10. Application of nanodiamonds in human body fluid analysis by matrix-assisted laser desorption/ionization mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    Xianglei Kong

    2008-01-01

    Direct mass spectrometric analysis of complex biological samples is very important and challenging. In this paper, nanodiamonds have been successfully used in matrix-assisted laser desorption/ionization mass spectrometric analysis of human serum and urine. As a practical tool and platform, it can be widely used in the field of humoral proteomics, and it plays a very promising role in clinical diagnosis, including identification of novel disease-associated biomarkers.

  11. Nanoparticle-assisted laser desorption/ionization mass spectrometry: Novel sample preparation methods and nanoparticle screening for plant metabolite imaging

    Energy Technology Data Exchange (ETDEWEB)

    Yagnik, Gargey B. [Iowa State Univ., Ames, IA (United States)

    2016-02-19

    The main goal of the presented research is development of nanoparticle based matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS). This dissertation includes the application of previously developed data acquisition methods, development of novel sample preparation methods, application and comparison of novel nanoparticle matrices, and comparison of two nanoparticle matrix application methods for MALDI-MS and MALDI-MS imaging.

  12. Comparative mass spectrometric analyses of Photofrin oligomers by fast atom bombardment mass spectrometry, UV and IR matrix-assisted laser desorption/ionization mass spectrometry, electrospray ionization mass spectrometry and laser desorption/jet-cooling photoionization mass spectrometry.

    Science.gov (United States)

    Siegel, M M; Tabei, K; Tsao, R; Pastel, M J; Pandey, R K; Berkenkamp, S; Hillenkamp, F; de Vries, M S

    1999-06-01

    Photofrin (porfimer sodium) is a porphyrin derivative used in the treatment of a variety of cancers by photodynamic therapy. This oligomer complex and a variety of porphyrin monomers, dimers and trimers were analyzed with five different mass spectral ionization techniques: fast atom bombardment, UV and IR matrix-assisted laser desorption/ionization, electrospray ionization, and laser desorption/jet-cooling photoionization. All five approaches resulted in very similar oligomer distributions with an average oligomer length of 2.7 +/- 0.1 porphyrin units. In addition to the Photofrin analysis, this study provides a side-by-side comparison of the spectra for the five different mass spectrometric techniques.

  13. Direct Analysis of Gold Nanoparticles from Dried Droplets Using Substrate-Assisted Laser Desorption Single Particle-ICPMS.

    Science.gov (United States)

    Benešová, Iva; Dlabková, Kristýna; Zelenák, František; Vaculovič, Tomáš; Kanický, Viktor; Preisler, Jan

    2016-03-01

    Single particle inductively coupled plasma mass spectrometry (SP-ICPMS) has been generally accepted as a powerful tool in the field of nanoanalysis. The method has usually been restricted to direct nanoparticle (NP) introduction using nebulization or microdroplet generation systems. In this work, AuNPs are introduced into ICPMS by substrate-assisted laser desorption (SALD) directly from a suitable absorbing plastic surface using a commercial ablation cell for the first time. In SALD, desorption of individual NPs is mediated using a frequency-quintupled Nd:YAG laser (213 nm) operated at a rather low laser fluence. Conditions including laser fluence, laser beam scan rate, and carrier gas flow rate were optimized in order to gain the highest AuNP transport efficiency and avoid AuNP disintegration within the laser irradiation. The method was demonstrated on a well-characterized reference material, 56 nm AuNPs with a transport efficiency of 61% and commercially available 86 nm AuNPs. Feasibility of our technique for NP detection and characterization is discussed here, and the results are compared with an established technique, nebulizer SP-ICPMS.

  14. Quantification of Carbohydrates and Related Materials Using Sodium Ion Adducts Produced by Matrix-Assisted Laser Desorption Ionization

    Science.gov (United States)

    Ahn, Sung Hee; Park, Kyung Man; Moon, Jeong Hee; Lee, Seong Hoon; Kim, Myung Soo

    2016-11-01

    The utility of sodium ion adducts produced by matrix-assisted laser desorption ionization for the quantification of analytes with multiple oxygen atoms was evaluated. Uses of homogeneous solid samples and temperature control allowed the acquisition of reproducible spectra. The method resulted in a direct proportionality between the ion abundance ratio I([A + Na]+)/I([M + Na]+) and the analyte concentration, which could be used as a calibration curve. This was demonstrated for carbohydrates, glycans, and polyether diols with dynamic range exceeding three orders of magnitude.

  15. Pigments and proteins in green bacterial chlorosomes studied by matrix-assisted laser desorption ionization mass spectrometry

    DEFF Research Database (Denmark)

    Persson, S; Sönksen, C P; Frigaard, N-U

    2000-01-01

    We have used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for mass determination of pigments and proteins in chlorosomes, the light-harvesting organelles from the photosynthetic green sulfur bacterium Chlorobium tepidum. By applying a small volume (1...... homologs in a small amount of green bacterial cells. In addition to information on pigments, the MALDI spectra also contained peaks from chlorosome proteins. Thus we have been able with high precision to confirm the molecular masses of the chlorosome proteins CsmA and CsmE which have been previously...

  16. Authenticity assessment of beef origin by principal component analysis of matrix-assisted laser desorption/ionization mass spectrometric data.

    Science.gov (United States)

    Zaima, Nobuhiro; Goto-Inoue, Naoko; Hayasaka, Takahiro; Enomoto, Hirofumi; Setou, Mitsutoshi

    2011-06-01

    It has become necessary to assess the authenticity of beef origin because of concerns regarding human health hazards. In this study, we used a metabolomic approach involving matrix-assisted laser desorption/ionization imaging mass spectrometry to assess the authenticity of beef origin. Highly accurate data were obtained for samples of extracted lipids from beef of different origin; the samples were grouped according to their origin. The analysis of extracted lipids in this study ended within 10 min, suggesting this approach can be used as a simple authenticity assessment before a definitive identification by isotope analysis.

  17. Current status of matrix-assisted laser desorption ionisation-time of flight mass spectrometry in the clinical microbiology laboratory.

    Science.gov (United States)

    Kok, Jen; Chen, Sharon C A; Dwyer, Dominic E; Iredell, Jonathan R

    2013-01-01

    The integration of matrix-assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF MS) into many clinical microbiology laboratories has revolutionised routine pathogen identification. MALDI-TOF MS complements and has good potential to replace existing phenotypic identification methods. Results are available in a more clinically relevant timeframe, particularly in bacteraemic septic shock. Novel applications include strain typing and the detection of antimicrobial resistance, but these are not widely used. This review discusses the technical aspects, current applications, and limitations of MALDI-TOF MS.

  18. Quantification of Carbohydrates and Related Materials Using Sodium Ion Adducts Produced by Matrix-Assisted Laser Desorption Ionization

    Science.gov (United States)

    Ahn, Sung Hee; Park, Kyung Man; Moon, Jeong Hee; Lee, Seong Hoon; Kim, Myung Soo

    2016-09-01

    The utility of sodium ion adducts produced by matrix-assisted laser desorption ionization for the quantification of analytes with multiple oxygen atoms was evaluated. Uses of homogeneous solid samples and temperature control allowed the acquisition of reproducible spectra. The method resulted in a direct proportionality between the ion abundance ratio I([A + Na]+)/I([M + Na]+) and the analyte concentration, which could be used as a calibration curve. This was demonstrated for carbohydrates, glycans, and polyether diols with dynamic range exceeding three orders of magnitude.

  19. Benefits of 2.94 μm infrared matrix-assisted laser desorption/ionization for analysis of labile molecules by Fourier transform mass spectrometry

    DEFF Research Database (Denmark)

    Budnik, Bogdan A.; Jensen, Kenneth Bendix; Jørgensen, Thomas J. D.

    2000-01-01

    A 2.94 microm Er:YAG laser was used together with a commercial Fourier transform mass spectrometer to study labile biomolecules. The combination has shown superior performance over conventional 337 nm ultraviolet matrix-assisted laser desorption/ionization (UV-MALDI) Fourier transform mass spectr...

  20. Ion-to-Neutral Ratios and Thermal Proton Transfer in Matrix-Assisted Laser Desorption/Ionization.

    Science.gov (United States)

    Lu, I-Chung; Chu, Kuan Yu; Lin, Chih-Yuan; Wu, Shang-Yun; Dyakov, Yuri A; Chen, Jien-Lian; Gray-Weale, Angus; Lee, Yuan-Tseh; Ni, Chi-Kung

    2015-07-01

    The ion-to-neutral ratios of four commonly used solid matrices, α-cyano-4-hydroxycinnamic acid (CHCA), 2,5-dihydroxybenzoic acid (2,5-DHB), sinapinic acid (SA), and ferulic acid (FA) in matrix-assisted laser desorption/ionization (MALDI) at 355 nm are reported. Ions are measured using a time-of-flight mass spectrometer combined with a time-sliced ion imaging detector. Neutrals are measured using a rotatable quadrupole mass spectrometer. The ion-to-neutral ratios of CHCA are three orders of magnitude larger than those of the other matrices at the same laser fluence. The ion-to-neutral ratios predicted using the thermal proton transfer model are similar to the experimental measurements, indicating that thermal proton transfer reactions play a major role in generating ions in ultraviolet-MALDI.

  1. Considerations for quantification of lipids in nerve tissue using matrix-assisted laser desorption/ionization mass spectrometric imaging.

    Science.gov (United States)

    Landgraf, Rachelle R; Garrett, Timothy J; Conaway, Maria C Prieto; Calcutt, Nigel A; Stacpoole, Peter W; Yost, Richard A

    2011-10-30

    Matrix-assisted laser desorption/ionization (MALDI) mass spectrometric imaging is a technique that provides the ability to identify and characterize endogenous and exogenous compounds spatially within tissue with relatively little sample preparation. While it is a proven methodology for qualitative analysis, little has been reported for its utility in quantitative measurements. In the current work, inherent challenges in MALDI quantification are addressed. Signal response is monitored over successive analyses of a single tissue section to minimize error due to variability in the laser, matrix application, and sample inhomogeneity. Methods for the application of an internal standard to tissue sections are evaluated and used to quantify endogenous lipids in nerve tissue. A precision of 5% or less standard error was achieved, illustrating that MALDI imaging offers a reliable means of in situ quantification for microgram-sized samples and requires minimal sample preparation.

  2. Recent Advances in Bacteria Identification by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Using Nanomaterials as Affinity Probes

    Directory of Open Access Journals (Sweden)

    Tai-Chia Chiu

    2014-04-01

    Full Text Available Identifying trace amounts of bacteria rapidly, accurately, selectively, and with high sensitivity is important to ensuring the safety of food and diagnosing infectious bacterial diseases. Microbial diseases constitute the major cause of death in many developing and developed countries of the world. The early detection of pathogenic bacteria is crucial in preventing, treating, and containing the spread of infections, and there is an urgent requirement for sensitive, specific, and accurate diagnostic tests. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS is an extremely selective and sensitive analytical tool that can be used to characterize different species of pathogenic bacteria. Various functionalized or unmodified nanomaterials can be used as affinity probes to capture and concentrate microorganisms. Recent developments in bacterial detection using nanomaterials-assisted MALDI-MS approaches are highlighted in this article. A comprehensive table listing MALDI-MS approaches for identifying pathogenic bacteria, categorized by the nanomaterials used, is provided.

  3. Furoic and mefenamic acids as new matrices for matrix assisted laser desorption/ionization-(MALDI)-mass spectrometry.

    Science.gov (United States)

    Abdelhamid, Hani Nasser; Wu, Hui-Fen

    2013-10-15

    The present study introduces two novel organic matrices for matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) for the analysis of small molecules. The first matrix is "2-amino-4,5-diphenylfuran-3-carboxylic acid" (also called furoic acid, FA) which was synthesized and then characterized by ultraviolet (UV), infrared (FTIR), nuclear magnetic resonance NMR ((1)H and (13)C) and mass spectrometry. The compound has organic semiconductor properties and exhibits intense UV-absorption which is suitable for the UV-MALDI laser (N2 laser, 337 nm). The second matrix is mefenamic acid (MA). The two matrices can be successfully applied for various classes of compounds including adenosine-5'-triphosphate (ATP, 0.5 µL(10.0 nmol)), spectinomycin (spect, 0.5 µL(14.0 nmol)), glutathione (GSH, 0.5 µL(9.0 nmol)), sulfamethazole (SMT, 0.5 µL(2.0 nmol)) and mixture of peptides gramicidin D (GD, 0.5µL (9.0 nmol)). The two matrices can effectively absorb the laser energy, resulting in excellent desorption/ionization of small molecules. The new matrices offer a significant enhancement of ionization, less fragmentation, few interferences, nice reproducibility, and excellent stability under vacuum. Theoretical calculations of the physical parameters demonstrated increase in polarizability, molar volume and refractivity than the conventional organic matrices which can effectively enhance the proton transfer reactions between the matrices with the analyte molecules. While the reduction in density, surface tension and index of refraction can enhance homogeneity between the two new matrices with the analytes. Due to the sublimation energy of mefenamic acid is (1.2 times) higher than that of the DHB, it is more stable to be used in the vacuum.

  4. Enhanced capabilities for imaging gangliosides in murine brain with matrix-assisted laser desorption/ionization and desorption electrospray ionization mass spectrometry coupled to ion mobility separation.

    Science.gov (United States)

    Škrášková, Karolina; Claude, Emmanuelle; Jones, Emrys A; Towers, Mark; Ellis, Shane R; Heeren, Ron M A

    2016-07-15

    The increased interest in lipidomics calls for improved yet simplified methods of lipid analysis. Over the past two decades, mass spectrometry imaging (MSI) has been established as a powerful technique for the analysis of molecular distribution of a variety of compounds across tissue surfaces. Matrix-assisted laser desorption/ionization (MALDI) MSI is widely used to study the spatial distribution of common lipids. However, a thorough sample preparation and necessity of vacuum for efficient ionization might hamper its use for high-throughput lipid analysis. Desorption electrospray ionization (DESI) is a relatively young MS technique. In DESI, ionization of molecules occurs under ambient conditions, which alleviates sample preparation. Moreover, DESI does not require the application of an external matrix, making the detection of low mass species more feasible due to the lack of chemical matrix background. However, irrespective of the ionization method, the final information obtained during an MSI experiment is very complex and its analysis becomes challenging. It was shown that coupling MSI to ion mobility separation (IMS) simplifies imaging data interpretation. Here we employed DESI and MALDI MSI for a lipidomic analysis of the murine brain using the same IMS-enabled instrument. We report for the first time on the DESI IMS-MSI of multiply sialylated ganglioside species, as well as their acetylated versions, which we detected directly from the murine brain tissue. We show that poly-sialylated gangliosides can be imaged as multiply charged ions using DESI, while they are clearly separated from the rest of the lipid classes based on their charge state using ion mobility. This represents a major improvement in MSI of intact fragile lipid species. We additionally show that complementary lipid information is reached under particular conditions when DESI is compared to MALDI MSI.

  5. Analysis of chlorophylls and their derivatives by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry.

    Science.gov (United States)

    Suzuki, Toshiyuki; Midonoya, Hitoshi; Shioi, Yuzo

    2009-07-01

    The analysis of chlorophylls and their derivatives by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry is described. Four matrices-sinapinic acid, a-cyano-4-hydroxycinnnamic acid, terthiophene, and 3-aminoquinoline-were examined to determine optimal conditions for analysis of the molecular mass and structure of chlorophyll a as a representative chlorophyll. Among them, terthiophene was the most efficient without releasing metal ions, although it caused fragmentation of the phytol-ester linkage. Terthiophene was useful for the analyses of chlorophyll derivatives as well as porphyrin products such as 8-deethyl-8-vinyl-chlorophyll a, pheophorbide a, pyropheophorbide a, bacteriochlorophyll a esterified phytol, and protoporphyrin IX. The current method is suitable for rapid and accurate determination of the molecular mass and structure of chlorophylls and porphyrins.

  6. Structural characterization of phospholipids by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry.

    Science.gov (United States)

    Marto, J A; White, F M; Seldomridge, S; Marshall, A G

    1995-11-01

    Matrix-assisted laser desorption/ionization (MALDI) Fourier transform ion cyclotron resonance mass spectrometry provides for structural analysis of the principal biological phospholipids: glycerophosphatidylcholine, -ethanolamine, -serine, and -inositol. Both positive and negative molecular or quasimolecular ions are generated in high abundance. Isolated molecular ions may be collisionally activated in the source side of a dual trap mass analyzer, yielding fragments serving to identify the polar head group (positive ion mode) and fatty acid side chains (negative ion mode). Azimuthal quadrupolar excitation following collisionally activated dissociation refocuses productions close to the solenoid axis; subsequent transfer of product ions to the analyzer ion trap allows for high-resolution mass analysis. Cyro-cooling of the sample probe with liquid nitrogen greatly reduces matrix adduction encountered in the negative ion mode.

  7. Matrix assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) for direct visualization of plant metabolites in situ.

    Science.gov (United States)

    Sturtevant, Drew; Lee, Young-Jin; Chapman, Kent D

    2016-02-01

    Direct visualization of plant tissues by matrix assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) has revealed key insights into the localization of metabolites in situ. Recent efforts have determined the spatial distribution of primary and secondary metabolites in plant tissues and cells. Strategies have been applied in many areas of metabolism including isotope flux analyses, plant interactions, and transcriptional regulation of metabolite accumulation. Technological advances have pushed achievable spatial resolution to subcellular levels and increased instrument sensitivity by several orders of magnitude. It is anticipated that MALDI-MSI and other MSI approaches will bring a new level of understanding to metabolomics as scientists will be encouraged to consider spatial heterogeneity of metabolites in descriptions of metabolic pathway regulation.

  8. Detection of Posaconazole by Surface-Assisted Laser Desorption/Ionization Mass Spectrometry with Dispersive Liquid-Liquid Microextraction

    Science.gov (United States)

    Lin, Sheng-Yu; Chen, Pin-Shiuan; Chang, Sarah Y.

    2015-03-01

    A simple, rapid, and sensitive method for the detection of posaconazole using dispersive liquid-liquid microextraction (DLLME) coupled to surface-assisted laser desorption/ionization mass spectrometric detection (SALDI/MS) was developed. After the DLLME, posaconazole was detected using SALDI/MS with colloidal gold and α-cyano-4-hydroxycinnamic acid (CHCA) as the co-matrix. Under optimal extraction and detection conditions, the calibration curve, which ranged from 1.0 to 100.0 nM for posaconazole, was observed to be linear. The limit of detection (LOD) at a signal-to-noise ratio of 3 was 0.3 nM for posaconazole. This novel method was successfully applied to the determination of posaconazole in human urine samples.

  9. Analysis and Quantitation of Glycated Hemoglobin by Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry

    Science.gov (United States)

    Hattan, Stephen J.; Parker, Kenneth C.; Vestal, Marvin L.; Yang, Jane Y.; Herold, David A.; Duncan, Mark W.

    2016-03-01

    Measurement of glycated hemoglobin is widely used for the diagnosis and monitoring of diabetes mellitus. Matrix assisted laser desorption/ionization (MALDI) time of flight (TOF) mass spectrometry (MS) analysis of patient samples is used to demonstrate a method for quantitation of total glycation on the β-subunit of hemoglobin. The approach is accurate and calibrated with commercially available reference materials. Measurements were linear (R2 > 0.99) across the clinically relevant range of 4% to 20% glycation with coefficients of variation of ≤ 2.5%. Additional and independent measurements of glycation of the α-subunit of hemoglobin are used to validate β-subunit glycation measurements and distinguish hemoglobin variants. Results obtained by MALDI-TOF MS were compared with those obtained in a clinical laboratory using validated HPLC methodology. MALDI-TOF MS sample preparation was minimal and analysis times were rapid making the method an attractive alternative to methodologies currently in practice.

  10. Affinity labeling coupled with matrix assistant laser desorption tandem time of flight mass spectrometry for quantitative proteomies research

    Institute of Scientific and Technical Information of China (English)

    MENG Qingfang; ZHANG Yangjun; CAI Yun; QIAN Xiaohong

    2007-01-01

    A relative quantitative method for differential proteomics by cleavable isotope-coded atTmity tag (cICAT)and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF-MS) was estab-lished. The accuracy and reproducibility of the method were evaluated by bovine serum albumin (BSA) digest as having a relative standard deviation of less than 30% and good reproducibility. The dynamic range was als0 evaluated by analyzing two mixtures of several standard proteins with dif-ferent concentration. The experimental results showed that in the dynamic range of 1:30, the quantitation error of the method was less than 30%. Although the quantitation error becomes very large when used beyond this range, it does not affect the derivation of information on the differential proteins. All the work provides an alternative method for differential proteomics analysis in biological samples from different origins.

  11. Analysis and Quantitation of Glycated Hemoglobin by Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry.

    Science.gov (United States)

    Hattan, Stephen J; Parker, Kenneth C; Vestal, Marvin L; Yang, Jane Y; Herold, David A; Duncan, Mark W

    2016-03-01

    Measurement of glycated hemoglobin is widely used for the diagnosis and monitoring of diabetes mellitus. Matrix assisted laser desorption/ionization (MALDI) time of flight (TOF) mass spectrometry (MS) analysis of patient samples is used to demonstrate a method for quantitation of total glycation on the β-subunit of hemoglobin. The approach is accurate and calibrated with commercially available reference materials. Measurements were linear (R(2) > 0.99) across the clinically relevant range of 4% to 20% glycation with coefficients of variation of ≤ 2.5%. Additional and independent measurements of glycation of the α-subunit of hemoglobin are used to validate β-subunit glycation measurements and distinguish hemoglobin variants. Results obtained by MALDI-TOF MS were compared with those obtained in a clinical laboratory using validated HPLC methodology. MALDI-TOF MS sample preparation was minimal and analysis times were rapid making the method an attractive alternative to methodologies currently in practice.

  12. Pigments and proteins in green bacterial chlorosomes studied by matrix-assisted laser desorption ionization mass spectrometry

    DEFF Research Database (Denmark)

    Persson, S; Sönksen, C P; Frigaard, N U;

    2000-01-01

    We have used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for mass determination of pigments and proteins in chlorosomes, the light-harvesting organelles from the photosynthetic green sulfur bacterium Chlorobium tepidum. By applying a small volume (1...... proportional to peak areas obtained from HPLC analysis of the same sample. The same result was also obtained when whole cells of Chl. tepidum were applied to the target, indicating that MALDI-MS can provide a rapid method for obtaining a semiquantitative determination or finger-print of the bacteriochlorophyll...... homologs in a small amount of green bacterial cells. In addition to information on pigments, the MALDI spectra also contained peaks from chlorosome proteins. Thus we have been able with high precision to confirm the molecular masses of the chlorosome proteins CsmA and CsmE which have been previously...

  13. Identification of Haemophilus influenzae and Haemophilus haemolyticus by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Bruin, J P; Kostrzewa, M; van der Ende, A; Badoux, P; Jansen, R; Boers, S A; Diederen, B M W

    2014-02-01

    Generally accepted laboratory methods that have been used for decades do not reliably distinguish between H. influenzae and H. haemolyticus isolates. H. haemolyticus strains are often incorrectly identified as nontypeable Haemophilus influenzae (NTHi). To distinguish H. influenzae from H. haemolyticus we have created a new database on the matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) bio-typer 2 and compared the results with routine determination of Haemophilus (growth requirement for X and V factor), and multilocus sequence typing (MLST). In total we have tested 277 isolates, 244 H. influenzae and 33 H. haemolyticus. Using MLST as the gold standard, the agreement of MALDI-TOF MS was 99.6 %. MALDI-TOF MS allows reliable and rapid discrimination between H. influenzae and H. haemolyticus.

  14. Natural products in Glycyrrhiza glabra (licorice) rhizome imaged at the cellular level by atmospheric pressure matrix-assisted laser desorption/ionization tandem mass spectrometry imaging

    DEFF Research Database (Denmark)

    Li, Bin; Bhandari, Dhaka Ram; Janfelt, Christian

    2014-01-01

    The rhizome of Glycyrrhiza glabra (licorice) was analyzed by high-resolution mass spectrometry imaging and tandem mass spectrometry imaging. An atmospheric pressure matrix-assisted laser desorption/ionization imaging ion source was combined with an orbital trapping mass spectrometer in order...

  15. Alkaloid profiling of the Chinese herbal medicine Fuzi by combination of matrix-assisted laser desorption ionization mass spectrometry with liquid chromatography-mass spectrometry

    NARCIS (Netherlands)

    Wang, J.; Heijden, R. van der; Spijksma, G.; Reijmers, T.; Wang, M.; Xu, G.; Hankemeier, T.; Greef, J. van der

    2009-01-01

    A matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) method was developed for the high throughput and robust qualitative profiling of alkaloids in Fuzi-the processed lateral roots of the Chinese herbal medicine Aconitum carmichaeli Debx (A. carmichaeli). After optimization, pow

  16. In situ liquid-liquid extraction as a sample preparation method for matrix-assisted laser desorption/ionization MS analysis of polypeptide mixtures

    DEFF Research Database (Denmark)

    Kjellström, Sven; Jensen, Ole Nørregaard

    2003-01-01

    A novel liquid-liquid extraction (LLE) procedure was investigated for preparation of peptide and protein samples for matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). LLE using ethyl acetate as the water-immiscible organic solvent enabled segregation of hydrophobic...

  17. Matrix-Assisted Laser Desorption Ionization Mass Spectrometry Imaging for Peptide and Protein Analyses: A Critical Review of On-Tissue Digestion

    NARCIS (Netherlands)

    Cillero-Pastor, B.; Heeren, R.M.A.

    2013-01-01

    Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has established itself among the plethora of mass spectrometry applications. In the biomedical field, MALDI-MSI is being more frequently recognized as a new method for the discovery of biomarkers and targets of treatme

  18. Differentiation of Raoultella ornithinolytica/planticola and Klebsiella oxytoca clinical isolates by matrix-assisted laser desorption/ionization-time of flight mass spectrometry.

    NARCIS (Netherlands)

    Jong, Eefje de; Jong, A.S. de; Smidts-van den Berg, N.; Rentenaar, R.J.

    2013-01-01

    Ninety-nine clinical isolates previously identified as Klebsiella oxytoca were evaluated using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Eight isolates were identified as Raoultella spp., being 5 Raoultella spp. and 3 K. oxytoca, by 16S rRNA sequenc

  19. Possible evidence of amide bond formation between sinapinic acid and lysine-containing bacterial proteins by matrix-assisted laser desorption/ionization (MALDI) at 355 nm

    Science.gov (United States)

    We previously reported the apparent formation of matrix adducts of 3,5-dimethoxy-4-hydroxy-cinnamic acid (sinapinic acid or SA) via covalent attachment to disulfide bond-containing proteins (HdeA, HdeB and YbgS) from bacterial cell lysates ionized by matrix-assisted laser desorption/ionization (MALD...

  20. Identification of Wheat Varieties Using Matrix-assisted Laser Desorption/Ionisation Time-of-flight Mass Spectrometry and an Artificial Neural network

    DEFF Research Database (Denmark)

    Bloch, Helle Aagaard; Kesmir, Can; Petersen, Marianne Kjerstine;

    1999-01-01

    A novel tool for variety identification of wheat (Triticum aestivum L,) has been developed: an artificial neural network (ANN) is used to classify the gliadin fraction analysed by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS). The robustness...

  1. Analysis of Phospholipid Mixtures from Biological Tissues by Matrix-Assisted Laser Desorption and Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS): A Laboratory Experiment

    Science.gov (United States)

    Eibisch, Mandy; Fuchs, Beate; Schiller, Jurgen; Sub, Rosmarie; Teuber, Kristin

    2011-01-01

    Matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) is increasingly used to investigate the phospholipid (PL) compositions of tissues and body fluids, often without previous separation of the total mixture into the individual PL classes. Therefore, the questions of whether all PL classes are detectable…

  2. Identification of Fatty Acids, Phospholipids, and Their Oxidation Products Using Matrix-Assisted Laser Desorption Ionization Mass Spectrometry and Electrospray Ionization Mass Spectrometry

    Science.gov (United States)

    Harmon, Christopher W.; Mang, Stephen A.; Greaves, John; Finlayson-Pitts, Barbara J.

    2010-01-01

    Electrospray ionization mass spectrometry (ESI-MS) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) have found increasing application in the analysis of biological samples. Using these techniques to solve problems in analytical chemistry should be an essential component of the training of undergraduate chemists. We…

  3. Differentiation of Clinically Relevant mucorales Rhizopus microsporus and R. arrhizus by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS)

    NARCIS (Netherlands)

    Dolatabadi, S.; Kolecka, A.; Versteeg, Matthijs; de Hoog, Sybren G; Boekhout, Teun

    2015-01-01

    This study addresses the usefulness of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) for reliable identification of the two most frequently occuring clinical species of Rhizopus, namely R. arrhizus with its two varieties arrhizus and delemar and R. micro

  4. Matrix-enhanced surface-assisted laser desorption/ionization mass spectrometry (ME-SALDI-MS) for mass spectrometry imaging of small molecules.

    Science.gov (United States)

    Brown, Victoria L; Liu, Qiang; He, Lin

    2015-01-01

    Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS), a parallel technique to matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), utilizes inorganic particles or porous surfaces to aid in the desorption/ionization of low-molecular-weight (MW) analytes. As a matrix-free and "soft" LDI approach, SALDI offers the benefit of reduced background noise in the low MW range, allowing for easier detection of biologically significant small MW species. Despite the inherent advantages of SALDI-MS, it has not reached comparable sensitivity levels to MALDI-MS. In relation to mass spectrometry imaging (MSI), intense efforts have been made in order to improve sensitivity and versatility of SALDI-MSI. We describe herein a detailed protocol that utilizes a hybrid LDI method, matrix-enhanced SALDI-MS (ME-SALDI MS), to detect and image low MW species in an imaging mode.

  5. Formation of aluminium, aluminium nitride and nitrogen clusters via laser ablation of nano aluminium nitride. Laser Desorption Ionisation and Matrix-Assisted Laser Desorption Ionisation Time-of-Flight Mass Spectrometry.

    Science.gov (United States)

    Panyala, Nagender Reddy; Prysiazhnyi, Vadym; Slavíček, Pavel; Černák, Mirko; Havel, Josef

    2011-06-30

    Laser Desorption Ionisation (LDI) and Matrix-Assisted Laser Desorption Ionisation (MALDI) Time-of-Flight Mass Spectrometry (TOFMS) were used to study the pulsed laser ablation of aluminium nitride (AlN) nano powder. The formation of Al(m)(+) (m=1-3), N(n)(+) (n=4, 5), AlN(n)(+) (n=1-5, 19, 21), Al(m)N(+) (m=2-3), Al(3)N(2)(+), Al(9)N(n)(+) (n=5, 7, 9, 11 and 15), Al(11)N(n)(+) (n=4, 6, 10, 12, 19, 21, 23, and 25), and Al(13)N(n)(+) (n=25, 31, 32, 33, 34, 35, and 36) clusters was detected in positive ion mode. Similarly, Al(m)(-) (m=1-3), AlN(n)(-) (n=1-3, 5), Al(m)N(-) (n=2, 3), Al(2)N(n)(-) (n=2-4, 28, 30), N(n)(-) (n=2, 3), Al(4)N(7)(-) Al(8)N(n)(-) (n=1-6), and Al(13)N(n)(-) (n=9, 18, 20, 22, 24, 26, 28, 33, 35, 37, 39, 41 and 43) clusters were observed in negative ion mode. The formation of the stoichiometric Al(10) N(10) cluster was shown to be of low abundance. On the contrary, the laser ablation of nano-AlN led mainly to the formation of nitrogen-rich Al(m)N(n) clusters in both negative and positive ion mode. The stoichiometry of the Al(m)N(n) clusters was determined via isotopic envelope analysis and computer modelling.

  6. Direct analysis of textile fabrics and dyes using infrared matrix-assisted laser desorption electrospray ionization mass spectrometry.

    Science.gov (United States)

    Cochran, Kristin H; Barry, Jeremy A; Muddiman, David C; Hinks, David

    2013-01-15

    The forensic analysis of textile fibers uses a variety of techniques from microscopy to spectroscopy. One such technique that is often used to identify the dye(s) within the fiber is mass spectrometry (MS). In the traditional MS method, the dye must be extracted from the fabric and the dye components are separated by chromatography prior to mass spectrometric analysis. Direct analysis of the dye from the fabric allows the omission of the lengthy sample preparation involved in extraction, thereby significantly reducing the overall analysis time. Herein, a direct analysis of dyed textile fabric was performed using the infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) source for MS. In MALDESI, an IR laser with wavelength tuned to 2.94 μm is used to desorb the dye from the fabric sample with the aid of water as the matrix. The desorbed dye molecules are then postionized by electrospray ionization (ESI). A variety of dye classes were analyzed from various fabrics with little to no sample preparation allowing for the identification of the dye mass and in some cases the fiber polymer. Those dyes that were not detected using MALDESI were also not observed by direct infusion ESI of the dye standard.

  7. Titanium dioxide anatase as matrix for matrix-assisted laser desorption/ionization analysis of small molecules.

    Science.gov (United States)

    Castro, Ana L; Madeira, Paulo J Amorim; Nunes, Manuel R; Costa, Fernanda M; Florêncio, M Helena

    2008-12-01

    The use of inorganic species as assisting materials in matrix-assisted laser desorption/ionization (MALDI) analysis is an alternative approach to avoid interfering matrix ions in the low-mass region of the mass spectra. Reports of the application of inorganic species as matrices in MALDI analysis of small molecules are, however, scarce. Nevertheless, titanium dioxide (TiO(2)) powder has been reported to be a promising matrix medium. In this study we further explore the use of TiO(2) as a matrix for the MALDI analysis of low molecular weight compounds. We present results showing that nanosized TiO(2) anatase and TiO(2) rutile perform better as MALDI matrices than a commercial TiO(2) anatase/rutile mixture. Moreover, when using nanosized TiO(2) anatase as a matrix, high-quality mass spectra can be obtained with strong analyte signals and weak or non-existing matrix interference ions. Furthermore, our results show that the phase type plays an important role in the application of TiO(2) as a MALDI matrix.

  8. Analysis of Microbial Mixtures by Matrix-assisted Laser Desorption/Ionization time-of-flight Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Wahl, Karen L.; Wunschel, Sharon C.; Jarman, Kristin H.; Valentine, Nancy B.; Petersen, Catherine E.; Kingsley, Mark T.; Zartolas, Kimberly A.; Saenz, Adam J.

    2002-12-15

    Many different laboratories are currently developing mass-spectrometric techniques to analyze and identify microorganisms. However, minimal work has been done with mixtures of bacteria. To demonstrate that microbial mixtures could be analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), mixed bacterial cultures were analyzed in a double-blind fashion. Nine different bacterial species currently in our MALDI-MS fingerprint library were used to generate 50 different simulated mixed bacterial cultures similar to that done for an initial blind study previously reported.(1) The samples were analyzed by MALDI-MS with automated data extraction and analysis algorithms developed in our laboratory. The components present in the sample were identified correctly to the species level in all but one of the samples. However, correctly eliminating closely related organisms was challenging for the current algorithms, especially in differentiating Serratia marcescens, Escherichia coli, and Yersinia enterocolitica, which have some similarities in their MALDI-MS fingerprints. Efforts to improve the specificity of the algorithms are in progress.

  9. Targeted comparative proteomics by liquid chromatography/matrix-assisted laser desorption/ionization triple-quadrupole mass spectrometry.

    Science.gov (United States)

    Melanson, Jeremy E; Chisholm, Kenneth A; Pinto, Devanand M

    2006-01-01

    Here we report the first application of a matrix-assisted laser desorption/ionization (MALDI) triple-quadrupole mass spectrometer for targeted proteomics. Employing an amine-specific isotopic labelling approach, the technique was validated using five randomly selected bovine serum albumin peptides differentially labelled at known ratios. An indirect benefit of the isotopic labelling technique is a significant enhancement of the a1 ion in tandem mass (MS/MS) spectra of all peptides studied. Therefore, the a1 ion was selected as the fragment ion for multiple reaction monitoring (MRM) in all cases, eliminating tedious method development and optimization. Accurate quantification was achieved with an average relative standard deviation (RSD) of 5% (n = 5) and a detection limit of 14 amol. The technique was then applied to validate an important virulence biomarker of the fungal pathogen Candida albicans, which was not accurately quantified using global proteomics experiment employing two-dimensional liquid chromatography/electrospray ionization tandem mass spectrometry (2D-LC/ESI)-MS/MS. Using LC/MALDI-MRM analysis of five tryptic peptides, the protein PHR1 was found to be upregulated in the hyphal (pathogenic) form of C. albicans by a factor of 7.7 +/- 0.8.

  10. Determination of paclitaxel distribution in solid tumors by nano-particle assisted laser desorption ionization mass spectrometry imaging.

    Directory of Open Access Journals (Sweden)

    Lavinia Morosi

    Full Text Available A sensitive, simple and reproducible protocol for nanoparticle-assisted laser desorption/ionization mass spectrometry imaging technique is described. The use of commercially available TiO2 nanoparticles abolishes heterogeneous crystallization, matrix background interferences and enhances signal detection, especially in the low mass range. Molecular image normalization was based on internal standard deposition on tissues, allowing direct comparison of drug penetration and distribution between different organs and tissues. The method was applied to analyze the distribution of the anticancer drug paclitaxel, inside normal and neoplastic mouse tissue sections. Spatial resolution was good, with a linear response between different in vivo treatments and molecular imaging intensity using therapeutic drug doses. This technique distinguishes the different intensity of paclitaxel distribution in control organs of mice, such as liver and kidney, in relation to the dose. Animals treated with 30 mg/kg of paclitaxel had half of the concentration of those treated with 60 mg/kg. We investigated the spatial distribution of paclitaxel in human melanoma mouse xenografts, following different dosage schedules and found a more homogeneous drug distribution in tumors of mice given repeated doses (5×8 mg/kg plus a 60 mg/kg dose than in those assigned only a single 60 mg/kg dose. The protocol can be readily applied to investigate anticancer drug distribution in neoplastic lesions and to develop strategies to optimize and enhance drug penetration through different tumor tissues.

  11. Direct analysis of pharmaceutical tablet formulations using Matrix-Assisted Laser Desorption/Ionisation Mass Spectrometry Imaging.

    Science.gov (United States)

    Earnshaw, Caroline J; Carolan, Vikki A; Richards, Don S; Clench, Malcolm R

    2010-06-15

    Matrix-Assisted Laser Desorption/Ionisation Mass Spectrometry Imaging (MALDI MSI) has been used to directly analyse a range of tablets in order to assess the homogeneity of the active drug compound throughout the excipients contained within the tablets studied. The information gained from the imaging experiments can be used to improve and gain a greater understanding of the manufacturing process; such knowledge will enable improvements in finished product quality to make safer and more efficacious tablet formulations. Commercially available and prescription tablet formulations have been analysed, including aspirin, paracetamol, sildenafil citrate (Viagra(R)) and a batch of tablets in development (tablet X: placebo; 1 mg; 3 mg and 6 mg). MALDI MSI provides semi-quantitative information that is related to ion abundance, therefore Principal Component Analysis (PCA), a multivariate analysis technique, has been used to differentiate between tablets containing different amounts of active drug ingredient. Aspects of sample preparation have also been investigated with regard to tablet shape and texture. The results obtained indicate that MALDI MSI can be used effectively to analyse the spatial distribution of the active pharmaceutical component (API) in pharmaceutical tablet formulations.

  12. Direct matrix-assisted laser desorption/ionization mass spectrometric imaging of cellulose and hemicellulose in Populus tissue.

    Science.gov (United States)

    Lunsford, Kyle Ann; Peter, Gary F; Yost, Richard A

    2011-09-01

    Imaging applied toward lignocellulosic materials requires high molecular specificity to map specific compounds within intact tissue. Although secondary ionization mass spectrometry (SIMS) and matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) with a single stage of MS have been used to image lignocellulosic biomass, the complexity of the plant tissue requires tandem MS, which limits the interpretation of simple MS. MALDI linear ion trap (LIT) tandem MS offers the high molecular specificity needed for lignocellulosic analyses. MALDI-LIT MS analyses of cellulose and xylan (hemicellulose) standards were performed to determine mass-to-charge ratios and fragmentation pathways for identification of these compounds in intact tissue. The MALDI-LIT-MS images of young Populus wood stem showed even distribution of both cellulose and hemicellulose ions; in contrast, the tandem MS images of cellulose and hemicellulose generated by plotting characteristic fragment ions resulted in drastically different images. This demonstrates that isobaric ions are present during MALDI-LIT-MS analyses of wood tissue and tandem MS is necessary to distinguish between isobaric species for selective imaging of carbohydrates in biomass.

  13. Matrix-assisted laser desorption-ionization time-of-flight mass spectrometry for the identification of Neisseria gonorrhoeae.

    Science.gov (United States)

    Buchanan, R; Ball, D; Dolphin, H; Dave, J

    2016-09-01

    Matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) was compared with the API NH biochemical method for the identification of Neisseria gonorrhoeae in routine clinical samples. A retrospective review of laboratory records for 1090 isolates for which both biochemical and MALDI-TOF MS identifications were available was performed. Cases of discrepant results were examined in detail for evidence supportive of a particular organism identification. Of 1090 isolates, 1082 were identified as N. gonorrhoeae by API NH. MALDI-TOF MS successfully identified 984 (91%) of these after one analysis, rising to 1081 (99.9%) after two analyses, with a positive predictive value of 99.3%. For those isolates requiring a repeat analysis, failure to generate an identifiable proteomic signature was the reason in 76% of cases, with alternative initial identifications accounting for the remaining 24%. MALDI-TOF MS identified eight isolates as N. gonorrhoeae that were not identified as such by API NH-examination of these discrepant results suggested that the MALDI-TOF MS identification may be the more reliable. MALDI-TOF MS is at least as accurate and reliable a method of identifying N. gonorrhoeae as API NH. We propose that MALDI-TOF MS could potentially be used as a single method for N. gonorrhoeae identification in routine cases by laboratories with access to this technology.

  14. Proteomic-based prognosis of brain tumor patients using direct-tissue matrix-assisted laser desorption ionization mass spectrometry.

    Science.gov (United States)

    Schwartz, Sarah A; Weil, Robert J; Thompson, Reid C; Shyr, Yu; Moore, Jason H; Toms, Steven A; Johnson, Mahlon D; Caprioli, Richard M

    2005-09-01

    Clinical diagnosis and treatment decisions for a subset of primary human brain tumors, gliomas, are based almost exclusively on tissue histology. Approaches for glioma diagnosis can be highly subjective due to the heterogeneity and infiltrative nature of these tumors and depend on the skill of the neuropathologist. There is therefore a critical need to develop more precise, non-subjective, and systematic methods to classify human gliomas. To this end, mass spectrometric analysis has been applied to these tumors to determine glioma-specific protein patterns. Protein profiles have been obtained from human gliomas of various grades through direct analysis of tissue samples using matrix-assisted laser desorption ionization mass spectrometry (MS). Statistical algorithms applied to the MS profiles from tissue sections identified protein patterns that correlated with tumor histology and patient survival. Using a data set of 108 glioma patients, two patient populations, a short-term and a long-term survival group, were identified based on the tissue protein profiles. In addition, a subset of 57 patients diagnosed with high-grade, grade IV, malignant gliomas were analyzed and a novel classification scheme that segregated short-term and long-term survival patients based on the proteomic profiles was developed. The protein patterns described served as an independent indicator of patient survival. These results show that this new molecular approach to monitoring gliomas can provide clinically relevant information on tumor malignancy and is suitable for high-throughput clinical screening.

  15. Principles of hydrogen radical mediated peptide/protein fragmentation during matrix-assisted laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Asakawa, Daiki

    2016-07-01

    Matrix-assisted laser desorption/ionization in-source decay (MALDI-ISD) is a very easy way to obtain large sequence tags and, thereby, reliable identification of peptides and proteins. Recently discovered new matrices have enhanced the MALDI-ISD yield and opened new research avenues. The use of reducing and oxidizing matrices for MALDI-ISD of peptides and proteins favors the production of fragmentation pathways involving "hydrogen-abundant" and "hydrogen-deficient" radical precursors, respectively. Since an oxidizing matrix provides information on peptide/protein sequences complementary to that obtained with a reducing matrix, MALDI-ISD employing both reducing and oxidizing matrices is a potentially useful strategy for de novo peptide sequencing. Moreover, a pseudo-MS(3) method provides sequence information about N- and C-terminus extremities in proteins and allows N- and C-terminal side fragments to be discriminated within the complex MALDI-ISD mass spectrum. The combination of high mass resolution of a Fourier transform-ion cyclotron resonance (FTICR) analyzer and the software suitable for MALDI-ISD facilitates the interpretation of MALDI-ISD mass spectra. A deeper understanding of the MALDI-ISD process is necessary to fully exploit this method. Thus, this review focuses first on the mechanisms underlying MALDI-ISD processes, followed by a discussion of MALDI-ISD applications in the field of proteomics. © 2014 Wiley Periodicals, Inc., Mass Spec Rev 35:535-556, 2016.

  16. Matrix-assisted laser desorption/ionization mass spectrometric analysis of aliphatic biodegradable photoluminescent polymers using new ionic liquid matrices.

    Science.gov (United States)

    Serrano, Carlos A; Zhang, Yi; Yang, Jian; Schug, Kevin A

    2011-05-15

    In this study, two novel ionic liquid matrices (ILMs), N,N-diisopropylethylammonium 3-oxocoumarate and N,N-diisopropylethylammonium dihydroxymonooxoacetophenoate, were tested for the structural elucidation of recently developed aliphatic biodegradable polymers by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The polymers, formed by a condensation reaction of three components, citric acid, octane diol, and an amino acid, are fluorescent, but the exact mechanism behind their luminescent properties has not been fully elucidated. In the original studies, which introduced the polymer class (J. Yang et al., Proc. Natl. Acad. Sci. USA 2009, 106, 10086-10091), a hyper-conjugated cyclic structure was proposed as the source for the photoluminescent behavior. With the use of the two new ILMs, we present evidence that supports the presence of the proposed cyclization product. In addition, the new ILMs, when compared with a previously established ILM, N,N-diisopropylethylammonium α-cyano-3-hydroxycinnimate, provided similar signal intensities and maintained similar spectral profiles. This research also established that the new ILMs provided good spot-to-spot reproducibility and high ionization efficiency compared with corresponding crystalline matrix preparations. Many polymer features revealed through the use of the ILMs could not be observed with crystalline matrices. Ultimately, the new ILMs highlighted the composition of the synthetic polymers, as well as the loss of water that was expected for the formation of the proposed cyclic structure on the polymer backbone.

  17. Matrix-assisted laser desorption ion trap mass spectrometry: efficient isolation and effective fragmentation of peptide ions.

    Science.gov (United States)

    Qin, J; Chait, B T

    1996-07-01

    Effective analysis of the sequence of peptides using matrix-assisted laser desorption/ionization (MALDI) tandem ion trap mass spectrometry requires efficient mass isolation and the ability to induce extensive sequence-specific fragmentation. The present paper describes a new excitation scheme, which we term red-shifted off-resonance large-amplitude excitation (RSORLAE), that can deposit higher amounts of internal energy in ions than is feasible with conventional resonant excitation. The new method provides an effective means for inducing fragmentation of MALDI-produced peptide ions with m/z values up to 3500. Prior to excitation, it is necessary to isolate ions of interest with high efficiency. We demonstrate that isolation efficiencies of > 95% can be achieved by careful design of the rf scan functions used during ion isolation. In particular, sudden transitions in the amplitude of the rf field (from low to high amplitudes) must be avoided. The combined improvements in the efficiency for ion isolation and the efficacy of ion activation make MALDI tandem ion trap mass spectrometry a practical tool for the characterization of proteins with high sensitivity.

  18. Identification and Classification of Rhizobia by Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry.

    Science.gov (United States)

    Jia, Rui Zong; Zhang, Rong Juan; Wei, Qing; Chen, Wen Feng; Cho, Il Kyu; Chen, Wen Xin; Li, Qing X

    Mass spectrometry (MS) has been widely used for specific, sensitive and rapid analysis of proteins and has shown a high potential for bacterial identification and characterization. Type strains of four species of rhizobia and Escherichia coli DH5α were employed as reference bacteria to optimize various parameters for identification and classification of species of rhizobia by matrix-assisted laser desorption/ionization time-of-flight MS (MALDI TOF MS). The parameters optimized included culture medium states (liquid or solid), bacterial growth phases, colony storage temperature and duration, and protein data processing to enhance the bacterial identification resolution, accuracy and reliability. The medium state had little effects on the mass spectra of protein profiles. A suitable sampling time was between the exponential phase and the stationary phase. Consistent protein mass spectral profiles were observed for E. coli colonies pre-grown for 14 days and rhizobia for 21 days at 4°C or 21°C. A dendrogram of 75 rhizobial strains of 4 genera was constructed based on MALDI TOF mass spectra and the topological patterns agreed well with those in the 16S rDNA phylogenetic tree. The potential of developing a mass spectral database for all rhizobia species was assessed with blind samples. The entire process from sample preparation to accurate identification and classification of species required approximately one hour.

  19. Matrix-assisted laser desorption/ionization mass spectrometry method for selectively producing either singly or multiply charged molecular ions.

    Science.gov (United States)

    Trimpin, Sarah; Inutan, Ellen D; Herath, Thushani N; McEwen, Charles N

    2010-01-01

    Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is noted for its ability to produce primarily singly charged ions. This is an attribute when using direct ionization for complex mixtures such as protein digests or synthetic polymers. However, the ability to produce multiply charged ions, as with electrospray ionization (ESI), has advantages such as extending the mass range on mass spectrometers with limited mass-to-charge (m/z) range and enhancing fragmentation for structural characterization. We designed and fabricated a novel field free transmission geometry atmopsheric pressure (AP) MALDI source mounted to a high-mass resolution Orbitrap Exactive mass spectrometer. We report the ability to produce at will either singly charged ions or highly charged ions using a MALDI process by simply changing the matrix or the matrix preparation conditions. Mass spectra with multiply charged ions very similar to those obtained with ESI of proteins such as cytochrome c and ubiquitin are obtained with low femtomole amounts applied to the MALDI target plate and for peptides such as angiotensin I and II with application of attomole amounts. Single scan acquisitions produce sufficient ion current even from proteins.

  20. Direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry improves appropriateness of antibiotic treatment of bacteremia.

    Science.gov (United States)

    Vlek, Anne L M; Bonten, Marc J M; Boel, C H Edwin

    2012-01-01

    Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows the identification of microorganisms directly from positive blood culture broths. Use of the MALDI-TOF MS for rapid identification of microorganisms from blood culture broths can reduce the turnaround time to identification and may lead to earlier appropriate treatment of bacteremia. During February and April 2010, direct MALDI-TOF MS was routinely performed on all positive blood cultures. During December 2009 and March 2010 no direct MALDI-TOF MS was used. Information on antibiotic therapy was collected from the hospital and intensive care units' information systems from all positive blood cultures during the study period. In total, 253 episodes of bacteremia were included of which 89 during the intervention period and 164 during the control period. Direct performance of MALDI-TOF MS on positive blood culture broths reduced the time till species identification by 28.8-h and was associated with an 11.3% increase in the proportion of patients receiving appropriate antibiotic treatment 24 hours after blood culture positivity (64.0% in the control period versus 75.3% in the intervention period (p0.01)). Routine implementation of this technique increased the proportion of patients on adequate antimicrobial treatment within 24 hours.

  1. Direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry improves appropriateness of antibiotic treatment of bacteremia.

    Directory of Open Access Journals (Sweden)

    Anne L M Vlek

    Full Text Available Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS allows the identification of microorganisms directly from positive blood culture broths. Use of the MALDI-TOF MS for rapid identification of microorganisms from blood culture broths can reduce the turnaround time to identification and may lead to earlier appropriate treatment of bacteremia. During February and April 2010, direct MALDI-TOF MS was routinely performed on all positive blood cultures. During December 2009 and March 2010 no direct MALDI-TOF MS was used. Information on antibiotic therapy was collected from the hospital and intensive care units' information systems from all positive blood cultures during the study period. In total, 253 episodes of bacteremia were included of which 89 during the intervention period and 164 during the control period. Direct performance of MALDI-TOF MS on positive blood culture broths reduced the time till species identification by 28.8-h and was associated with an 11.3% increase in the proportion of patients receiving appropriate antibiotic treatment 24 hours after blood culture positivity (64.0% in the control period versus 75.3% in the intervention period (p0.01. Routine implementation of this technique increased the proportion of patients on adequate antimicrobial treatment within 24 hours.

  2. Determination of Macrolide Antibiotics Using Dispersive Liquid-Liquid Microextraction Followed by Surface-Assisted Laser Desorption/Ionization Mass Spectrometry

    Science.gov (United States)

    Chen, Kuan-Yu; Yang, Thomas C.; Chang, Sarah Y.

    2012-06-01

    A novel method for the determination of macrolide antibiotics using dispersive liquid-liquid microextraction coupled to surface-assisted laser desorption/ionization mass spectrometric detection was developed. Acetone and dichloromethane were used as the disperser solvent and extraction solvent, respectively. A mixture of extraction solvent and disperser solvent were rapidly injected into a 1.0 mL aqueous sample to form a cloudy solution. After the extraction, macrolide antibiotics were detected using surface-assisted laser desorption/ionization mass spectrometry (SALDI/MS) with colloidal silver as the matrix. Under optimum conditions, the limits of detection (LODs) at a signal-to-noise ratio of 3 were 2, 3, 3, and 2 nM for erythromycin (ERY), spiramycin (SPI), tilmicosin (TILM), and tylosin (TYL), respectively. This developed method was successfully applied to the determination of macrolide antibiotics in human urine samples.

  3. Simplified sample preparation method for protein identification by matrix-assisted laser desorption/ionization mass spectrometry: in-gel digestion on the probe surface

    DEFF Research Database (Denmark)

    Stensballe, A; Jensen, Ole Nørregaard

    2001-01-01

    Identification and detailed characterization of complex mixtures of proteins separated by polyacrylamide gel electrophoresis (PAGE) require optimized and robust methods for interfacing electrophoretic techniques to mass spectrometry. Peptide mapping by matrix-assisted laser desorption/ionization-......Identification and detailed characterization of complex mixtures of proteins separated by polyacrylamide gel electrophoresis (PAGE) require optimized and robust methods for interfacing electrophoretic techniques to mass spectrometry. Peptide mapping by matrix-assisted laser desorption...... for protein identification similar to that obtained by the traditional protocols for in-gel digestion and MALDI peptide mass mapping of human proteins, i.e. approximately 60%. The overall performance of the novel on-probe digestion method is comparable with that of the standard in-gel sample preparation...

  4. Characterization of some synthetic Ru and Ir complexes by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    NARCIS (Netherlands)

    Lou, X.; Buijtenen, J. van; Bastiaansen, J.J.A.M.; Waal, B.F.M. de; Langeveld, B.M.W.; Dongen, J.L.J. van

    2005-01-01

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was applied to the analysis of Ru(OCOCF 3)2(CO)(PPh3)2, Ru(OCOC 3F7)2(CO)(PPh3)2, Ir(tBuppy)3 and Ir(ppy)2(acac) complexes. A troublesome problem in the MALDI-TOFMS characterization of these metal complexes is

  5. Identification of non-diphtheriae corynebacterium by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Alatoom, Adnan A; Cazanave, Charles J; Cunningham, Scott A; Ihde, Sherry M; Patel, Robin

    2012-01-01

    We evaluated the Bruker Biotyper matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry for identification of 92 clinical isolates of Corynebacterium species in comparison to identification using rpoB or 16S rRNA gene sequencing. Eighty isolates (87%) yielded a score of ≥1.700, and all of these were correctly identified to the species level with the exception of Corynebacterium aurimucosum being misidentified as the closely related Corynebacterium minutissimum.

  6. Assessment of Reproducibility of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Bacterial and Yeast Identification

    OpenAIRE

    Westblade, Lars F.; Garner, Omai B.; MacDonald,Karen; Bradford, Constance; Pincus, David H.; Mochon, A. Brian; Jennemann, Rebecca; Manji, Ryhana; Bythrow, Maureen; Lewinski, Michael A.; Burnham, Carey-Ann D.; Ginocchio, Christine C.

    2015-01-01

    Matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry (MS) has revolutionized the identification of clinical bacterial and yeast isolates. However, data describing the reproducibility of MALDI-TOF MS for microbial identification are scarce. In this study, we show that MALDI-TOF MS-based microbial identification is highly reproducible and can tolerate numerous variables, including differences in testing environments, instruments, operators, reagent lots, and ...

  7. Differentiation of Raoultella ornithinolytica/planticola and Klebsiella oxytoca clinical isolates by matrix-assisted laser desorption/ionization-time of flight mass spectrometry.

    Science.gov (United States)

    de Jong, Eefje; de Jong, Arjan S; Smidts-van den Berg, Nathalie; Rentenaar, Rob J

    2013-04-01

    Ninety-nine clinical isolates previously identified as Klebsiella oxytoca were evaluated using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Eight isolates were identified as Raoultella spp., being 5 Raoultella spp. and 3 K. oxytoca, by 16S rRNA sequencing. These isolates were correctly identified by applying the 10% differential rule for the MALDI-TOF MS score values. This approach might be useful to discriminate Raoultella species from K. oxytoca.

  8. Influence of Culture Media on Detection of Carbapenem Hydrolysis by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

    Science.gov (United States)

    Ramos, Ana Carolina; Carvalhaes, Cecília Godoy; Cordeiro-Moura, Jhonatha Rodrigo; Rockstroh, Anna Carolina; Machado, Antonia Maria Oliveira; Gales, Ana Cristina

    2016-07-01

    In this study, we evaluated the influence of distinct bacterial growth media on detection of carbapenemase hydrolysis by matrix-assisted laser desorption ionization-time of flight mass spectrometry. False-negative results were observed for OXA-25-, OXA-26-, and OXA-72-producing Acinetobacter baumannii isolates grown on MacConkey agar medium. The other culture media showed 100% sensitivity and 100% specificity for detecting carbapenemase.

  9. Rapid Identification of the Foodborne Pathogen Trichinella spp. by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.

    Directory of Open Access Journals (Sweden)

    Anne Mayer-Scholl

    Full Text Available Human trichinellosis occurs through consumption of raw or inadequately processed meat or meat products containing larvae of the parasitic nematodes of the genus Trichinella. Currently, nine species and three genotypes are recognized, of which T. spiralis, T. britovi and T. pseudospiralis have the highest public health relevance. To date, the differentiation of the larvae to the species and genotype level is based primarily on molecular methods, which can be relatively time consuming and labor intensive. Due to its rapidness and ease of use a matrix assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF MS reference spectra database using Trichinella strains of all known species and genotypes was created. A formicacid/acetonitrile protein extraction was carried out after pooling 10 larvae of each Trichinella species and genotype. Each sample was spotted 9 times using α-cyano 4-hydoxy cinnamic acid matrix and a MicroFlex LT mass spectrometer was used to acquire 3 spectra (m/z 2000 to 20000 Da from each spot resulting in 27 spectra/species or genotype. Following the spectra quality assessment, Biotyper software was used to create a main spectra library (MSP representing nine species and three genotypes of Trichinella. The evaluation of the spectra generated by MALDI-TOF MS revealed a classification which was comparable to the results obtained by molecular methods. Also, each Trichinella species utilized in this study was distinct and distinguishable with a high confidence level. Further, different conservation methods such as freezing and conservation in alcohol and the host species origin of the isolated larvae did not have a significant influence on the generated spectra. Therefore, the described MALDI-TOF MS can successfully be implemented for both genus and species level identification and represents a major step forward in the use of this technique in foodborne parasitology.

  10. Identification of beer-spoilage bacteria using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Wieme, Anneleen D; Spitaels, Freek; Aerts, Maarten; De Bruyne, Katrien; Van Landschoot, Anita; Vandamme, Peter

    2014-08-18

    Applicability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for identification of beer-spoilage bacteria was examined. To achieve this, an extensive identification database was constructed comprising more than 4200 mass spectra, including biological and technical replicates derived from 273 acetic acid bacteria (AAB) and lactic acid bacteria (LAB), covering a total of 52 species, grown on at least three growth media. Sequence analysis of protein coding genes was used to verify aberrant MALDI-TOF MS identification results and confirmed the earlier misidentification of 34 AAB and LAB strains. In total, 348 isolates were collected from culture media inoculated with 14 spoiled beer and brewery samples. Peak-based numerical analysis of MALDI-TOF MS spectra allowed a straightforward species identification of 327 (94.0%) isolates. The remaining isolates clustered separately and were assigned through sequence analysis of protein coding genes either to species not known as beer-spoilage bacteria, and thus not present in the database, or to novel AAB species. An alternative, classifier-based approach for the identification of spoilage bacteria was evaluated by combining the identification results obtained through peak-based cluster analysis and sequence analysis of protein coding genes as a standard. In total, 263 out of 348 isolates (75.6%) were correctly identified at species level and 24 isolates (6.9%) were misidentified. In addition, the identification results of 50 isolates (14.4%) were considered unreliable, and 11 isolates (3.2%) could not be identified. The present study demonstrated that MALDI-TOF MS is well-suited for the rapid, high-throughput and accurate identification of bacteria isolated from spoiled beer and brewery samples, which makes the technique appropriate for routine microbial quality control in the brewing industry.

  11. Rapid screening of mixed edible oils and gutter oils by matrix-assisted laser desorption/ionization mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Ng, Tsz-Tsun; So, Pui-Kin; Zheng, Bo [Food Safety and Technology Research Centre, State Key Laboratory of Chirosciences and Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom Kowloon, Hong Kong Special Administrative Region (China); Shenzhen Key Laboratory of Food Biological Safety Control and State Key Laboratory of Chinese Medicine and Molecular Pharmacology (Incubation), Shenzhen Research Institute of The Hong Kong Polytechnic University, Shenzhen (China); Yao, Zhong-Ping, E-mail: zhongping.yao@polyu.edu.hk [Food Safety and Technology Research Centre, State Key Laboratory of Chirosciences and Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom Kowloon, Hong Kong Special Administrative Region (China); Shenzhen Key Laboratory of Food Biological Safety Control and State Key Laboratory of Chinese Medicine and Molecular Pharmacology (Incubation), Shenzhen Research Institute of The Hong Kong Polytechnic University, Shenzhen (China)

    2015-07-16

    Highlights: • Simplified sample preparation method for direct analysis of edible oils by MALDI-MS. • Establishment of a preliminary MALDI-MS spectral database of edible oils. • Rapid screening of mixed edible oils and gutter oils. - Abstract: Authentication of edible oils is a long-term issue in food safety, and becomes particularly important with the emergence and wide spread of gutter oils in recent years. Due to the very high analytical demand and diversity of gutter oils, a high throughput analytical method and a versatile strategy for authentication of mixed edible oils and gutter oils are highly desirable. In this study, an improved matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) method has been developed for direct analysis of edible oils. This method involved on-target sample loading, automatic data acquisition and simple data processing. MALDI-MS spectra with high quality and high reproducibility have been obtained using this method, and a preliminary spectral database of edible oils has been set up. The authenticity of an edible oil sample can be determined by comparing its MALDI-MS spectrum and principal component analysis (PCA) results with those of its labeled oil in the database. This method is simple and the whole process only takes several minutes for analysis of one oil sample. We demonstrated that the method was sensitive to change in oil compositions and can be used for measuring compositions of mixed oils. The capability of the method for determining mislabeling enables it for rapid screening of gutter oils since fraudulent mislabeling is a common feature of gutter oils.

  12. Localization of ergot alkaloids in sclerotia of Claviceps purpurea by matrix-assisted laser desorption/ionization mass spectrometry imaging.

    Science.gov (United States)

    Dopstadt, Julian; Vens-Cappell, Simeon; Neubauer, Lisa; Tudzynski, Paul; Cramer, Benedikt; Dreisewerd, Klaus; Humpf, Hans-Ulrich

    2017-02-01

    The fungus Claviceps purpurea produces highly toxic ergot alkaloids and accumulates these in the hardened bodies of fungal mycelium. These so-called sclerotia, or ergot bodies, replace the crop seed of infected plants, which can include numerous important food- and feedstuff such as rye and wheat. While several studies have explored details of the infection process and development of ergot bodies, little information is available on the spatial distribution of the mycotoxins in the sclerotia. Here we used matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) at a lateral resolution of 35 μm to visualize the distribution of two representative alkaloids, ergocristine and ergometrine, produced by Ecc93 and Gal 310 variants of C. purpurea, respectively, after infection of rye. To improve cryosectioning of this fragile biological material tissue with complex texture, we developed a practical embedding protocol based on cellulose polymers. The MALDI-MS images recorded from the so produced intact tissues sections revealed that ergometrine exhibited a relatively homogeneous distribution throughout the ergot body, whereas ergocristine was found to be enriched in the proximal region. This finding can be correlated to the morphological development of sclerotia as ergot alkaloids are only produced in the sphacelial stage. The ability to localize toxins and other secondary metabolites in intact sections of crop-infecting fungi with high lateral resolution renders MALDI-MSI a powerful tool for investigating biosynthetic pathways and for obtaining a deeper understanding of the parasite-host interaction. Graphical abstract Workflow for identification and spatial localization of ergot alkaloids in infected rye grains.

  13. Negative Ion In-Source Decay Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry for Sequencing Acidic Peptides

    Science.gov (United States)

    McMillen, Chelsea L.; Wright, Patience M.; Cassady, Carolyn J.

    2016-05-01

    Matrix-assisted laser desorption/ionization (MALDI) in-source decay was studied in the negative ion mode on deprotonated peptides to determine its usefulness for obtaining extensive sequence information for acidic peptides. Eight biological acidic peptides, ranging in size from 11 to 33 residues, were studied by negative ion mode ISD (nISD). The matrices 2,5-dihydroxybenzoic acid, 2-aminobenzoic acid, 2-aminobenzamide, 1,5-diaminonaphthalene, 5-amino-1-naphthol, 3-aminoquinoline, and 9-aminoacridine were used with each peptide. Optimal fragmentation was produced with 1,5-diaminonphthalene (DAN), and extensive sequence informative fragmentation was observed for every peptide except hirudin(54-65). Cleavage at the N-Cα bond of the peptide backbone, producing c' and z' ions, was dominant for all peptides. Cleavage of the N-Cα bond N-terminal to proline residues was not observed. The formation of c and z ions is also found in electron transfer dissociation (ETD), electron capture dissociation (ECD), and positive ion mode ISD, which are considered to be radical-driven techniques. Oxidized insulin chain A, which has four highly acidic oxidized cysteine residues, had less extensive fragmentation. This peptide also exhibited the only charged localized fragmentation, with more pronounced product ion formation adjacent to the highly acidic residues. In addition, spectra were obtained by positive ion mode ISD for each protonated peptide; more sequence informative fragmentation was observed via nISD for all peptides. Three of the peptides studied had no product ion formation in ISD, but extensive sequence informative fragmentation was found in their nISD spectra. The results of this study indicate that nISD can be used to readily obtain sequence information for acidic peptides.

  14. Rapid Identification of the Foodborne Pathogen Trichinella spp. by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.

    Science.gov (United States)

    Mayer-Scholl, Anne; Murugaiyan, Jayaseelan; Neumann, Jennifer; Bahn, Peter; Reckinger, Sabine; Nöckler, Karsten

    2016-01-01

    Human trichinellosis occurs through consumption of raw or inadequately processed meat or meat products containing larvae of the parasitic nematodes of the genus Trichinella. Currently, nine species and three genotypes are recognized, of which T. spiralis, T. britovi and T. pseudospiralis have the highest public health relevance. To date, the differentiation of the larvae to the species and genotype level is based primarily on molecular methods, which can be relatively time consuming and labor intensive. Due to its rapidness and ease of use a matrix assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF MS) reference spectra database using Trichinella strains of all known species and genotypes was created. A formicacid/acetonitrile protein extraction was carried out after pooling 10 larvae of each Trichinella species and genotype. Each sample was spotted 9 times using α-cyano 4-hydoxy cinnamic acid matrix and a MicroFlex LT mass spectrometer was used to acquire 3 spectra (m/z 2000 to 20000 Da) from each spot resulting in 27 spectra/species or genotype. Following the spectra quality assessment, Biotyper software was used to create a main spectra library (MSP) representing nine species and three genotypes of Trichinella. The evaluation of the spectra generated by MALDI-TOF MS revealed a classification which was comparable to the results obtained by molecular methods. Also, each Trichinella species utilized in this study was distinct and distinguishable with a high confidence level. Further, different conservation methods such as freezing and conservation in alcohol and the host species origin of the isolated larvae did not have a significant influence on the generated spectra. Therefore, the described MALDI-TOF MS can successfully be implemented for both genus and species level identification and represents a major step forward in the use of this technique in foodborne parasitology.

  15. Large molecular mass materials in coal derived liquids by {sup 252}Cf-plasma and matrix assisted laser desorption mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Domin, M. [School of Pharmacy, London (United Kingdom). Dept. of Pharmaceutical and Biological Chemistry; Li, S.; Herod, A.A.; Larsen, J.W. [Lehigh Univ., Bethlehem, PA (United States). Dept. of Chemistry; Lazaro, M.J.; Kandiyoti, R. [Imperial College, London (United Kingdom). Dept. of Chemical Engineering and Chemical Technology

    1997-12-31

    A Point of Ayr coal extract, its hydrocracked product and the pyridine solubles/insolubles of a coal tar pitch have been examined using {sup 252}Cf-plasma desorption-mass spectrometry (PDMS) and matrix assisted laser desorption-mass spectrometry (MALDI-MS). Comparison of molecular masses (MMs) between the coal extract and its hydrocracked product by PDMS indicated ranges of masses in the product to be considerably smaller, with number and weight average MMs reduced by approximately a factor of two. MALDI-mass spectra of the same samples indicated a greater reduction in mass. Similar comparison of the pyridine soluble/insoluble fractions of the coal tar pitch showed smaller differences by PD-MS than by MALDI-MS. (orig.)

  16. Evaluation of matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry for identification of Candida parapsilosis, C. orthopsilosis and C. metapsilosis.

    Science.gov (United States)

    Quiles-Melero, I; García-Rodríguez, J; Gómez-López, A; Mingorance, J

    2012-01-01

    We have evaluated matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry for the rapid identification of Candida parapsilosis, C. orthopsilosis and C. metapsilosis. A total of 103 isolates, including reference strains and clinical isolates, were identified by pyrosequencing of the ITS1 region and then assay by MALDI-TOF mass spectrometry. Concordance between the two methods was 100%, showing that MALDI-TOF may be useful as a rapid and reliable method for discrimination of species within the C. parapsilosis group.

  17. A novel type of matrix for surface-assisted laser desorption-ionization mass spectrometric detection of biomolecules using metal-organic frameworks.

    Science.gov (United States)

    Fu, Chien-Ping; Lirio, Stephen; Liu, Wan-Ling; Lin, Chia-Her; Huang, Hsi-Ya

    2015-08-12

    A 3D metal-organic framework (MOF) nanomaterial as matrix for surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) and tandem mass spectrometry (MS/MS) was developed for the analysis of complex biomolecules. Unlike other nanoparticle matrices, this MOF nanomaterial does not need chemical modification prior to use. An exceptional signal reproducibility as well as very low background interferences in analyzing mono-/di-saccharides, peptides and complex starch digests demonstrate its high potential for biomolecule assays, especially for small molecules.

  18. Evaluation of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Identification of KPC-Producing Klebsiella pneumoniae.

    Science.gov (United States)

    Gaibani, Paolo; Galea, Anna; Fagioni, Marco; Ambretti, Simone; Sambri, Vittorio; Landini, Maria Paola

    2016-10-01

    We evaluated a real-time single-peak (11.109-Da) detection assay based on matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for the identification of Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae Our results demonstrated that the 11.109-Da peak was detected in 88.2% of the KPC producers. Analysis of blaKPC-producing K. pneumoniae showed that the gene encoding the 11.109-Da protein was commonly (97.8%) associated with the Tn4401a isoform.

  19. Evaluation of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Identification of KPC-Producing Klebsiella pneumoniae

    Science.gov (United States)

    Galea, Anna; Fagioni, Marco; Ambretti, Simone; Sambri, Vittorio; Landini, Maria Paola

    2016-01-01

    We evaluated a real-time single-peak (11.109-Da) detection assay based on matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) for the identification of Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae. Our results demonstrated that the 11.109-Da peak was detected in 88.2% of the KPC producers. Analysis of blaKPC-producing K. pneumoniae showed that the gene encoding the 11.109-Da protein was commonly (97.8%) associated with the Tn4401a isoform. PMID:27413192

  20. Assessment of Reproducibility of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Bacterial and Yeast Identification.

    Science.gov (United States)

    Westblade, Lars F; Garner, Omai B; MacDonald, Karen; Bradford, Constance; Pincus, David H; Mochon, A Brian; Jennemann, Rebecca; Manji, Ryhana; Bythrow, Maureen; Lewinski, Michael A; Burnham, Carey-Ann D; Ginocchio, Christine C

    2015-07-01

    Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has revolutionized the identification of clinical bacterial and yeast isolates. However, data describing the reproducibility of MALDI-TOF MS for microbial identification are scarce. In this study, we show that MALDI-TOF MS-based microbial identification is highly reproducible and can tolerate numerous variables, including differences in testing environments, instruments, operators, reagent lots, and sample positioning patterns. Finally, we reveal that samples of bacterial and yeast isolates prepared for MALDI-TOF MS identification can be repeatedly analyzed without compromising organism identification.

  1. Semiconductor Nanomaterials-Based Fluorescence Spectroscopic and Matrix-Assisted Laser Desorption/Ionization (MALDI Mass Spectrometric Approaches to Proteome Analysis

    Directory of Open Access Journals (Sweden)

    Suresh Kumar Kailasa

    2013-12-01

    Full Text Available Semiconductor quantum dots (QDs or nanoparticles (NPs exhibit very unusual physico-chemcial and optical properties. This review article introduces the applications of semiconductor nanomaterials (NMs in fluorescence spectroscopy and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS for biomolecule analysis. Due to their unique physico-chemical and optical properties, semiconductors NMs have created many new platforms for investigating biomolecular structures and information in modern biology. These semiconductor NMs served as effective fluorescent probes for sensing proteins and cells and acted as affinity or concentrating probes for enriching peptides, proteins and bacteria proteins prior to MALDI-MS analysis.

  2. Structure Determination of β-Glucans from Ganoderma lucidum with Matrix-assisted Laser Desorption/ionization (MALDI Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Wen-Bin Yang

    2008-08-01

    Full Text Available A novel method that uses matrix-assisted laser desorption/ionization (MALDI mass spectrometry to analyze molecular weight and sequencing of glucan in Ganoderma lucidum is presented. Thus, β-glucan, which was isolated from fruiting bodies of G. lucidum, was measured in a direct and fast way using MALDI mass spectrometry. In addition, tandem mass spectrometry of permethylated glucans of G. lucidum, dextran, curdlan and maltohexaose were also pursued and different fragment patterns were obtained. The G. lucidum glucan structure was determined and this method for linkage analysis of permethylated glucan has been proven feasible.

  3. Quantitation of peptides and proteins by matrix-assisted laser desorption/ionization mass spectrometry using (18)O-labeled internal standards

    DEFF Research Database (Denmark)

    Mirgorodskaya, O A; Kozmin, Y P; Titov, M I;

    2000-01-01

    A method for quantitating proteins and peptides in the low picomole and sub-picomole range has been developed using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) with internal (18)O-labeled standards. A simple procedure is proposed to produce such internal standards...... for the tested sample by enzymatic hydrolysis of the same sample (with known concentration) in (18)O-water. A mathematical algorithm was developed which uses the isotopic patterns of the substance, the internal standard, and the substance/internal standard mixture for accurate quantitation of the substance...

  4. Large molecular mass materials in coal-derived liquids by {sup 252}Cf-plasma and matrix-assisted laser desorption mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Domin, M.; Li, S.; Lazaro, M.-J.; Herod, A.A.; Larsen, J.W.; Kandiyoti, R. [School of Pharmacy, London (United Kingdom). Dept. of Parmaceutical and Biological Chemistry

    1998-05-01

    The paper compares responses of {sup 252}Cf-plasma desorption MS (PD-MS) and matrix-assisted laser desorption (MALDI) MS to identical samples. The two pairs of samples selected for the comparison were known from previous work to differ significantly in their high mass contents. MALDI-MS showed large differences in MM distributions within both pairs of samples. The PD-MS data showed a degree of similarity between one pair of samples (pyridine soluble/insoluble fractions of a coal tar pitch); for the second pair (a coal extract and its hydrocracked product), trends from the two MS techniques agreed closely. The MM range observed by PD-MS was somewhat narrower, extending to between 3000 and 5000 u. Significant differences within pairs of samples were observed by SEC and by UV-fluorescence spectroscopy, providing somewhat closer agreement with the MALDI spectra. The two MS instruments differ in two important respects: the ionization system (i.e., plasma vs laser desorption) and the maximum available ion extraction voltage: 30 kV for the MALDI-MS instrument and 15 kV for the PD-MS. The comparison of plasma vs laser desorption mass spectroscopy could not therefore take place at high ion extraction voltages. Work at up to 30 kV in the MALDI instrument indicated better sensitivity to high-mass materials at higher ion extraction voltages. The qualitative similarity of results from the two MS techniques is nevertheless apparent; the range of MMs observed in PD-MS as well as in MALDI-MS were, furthermore, far larger than those reported by any MS technique, to date. 38 refs., 6 figs., 1 tab.

  5. Direct Imaging Mass Spectrometry of Plant Leaves Using Surface-assisted Laser Desorption/Ionization with Sputter-deposited Platinum Film.

    Science.gov (United States)

    Ozawa, Tomoyuki; Osaka, Issey; Hamada, Satoshi; Murakami, Tatsuya; Miyazato, Akio; Kawasaki, Hideya; Arakawa, Ryuichi

    2016-01-01

    Plant leaves administered with systemic insecticides as agricultural chemicals were analyzed using imaging mass spectrometry (IMS). Matrix-assisted laser desorption/ionization (MALDI) is inadequate for the detection of insecticides on leaves because of the charge-up effect that occurs on the non-conductive surface of the leaves. In this study, surface-assisted laser desorption/ionization with a sputter-deposited platinum film (Pt-SALDI) was used for direct analysis of chemicals in plant leaves. Sputter-deposited platinum (Pt) films were prepared on leaves administered with the insecticides. A sputter-deposited Pt film with porous structure was used as the matrix for Pt-SALDI. Acephate and acetamiprid contained in the insecticides on the leaves could be detected using Pt-SALDI-MS, but these chemical components could not be adequately detected using MALDI-MS because of the charge-up effect. Enhancement of ion yields for the insecticides was achieved using Pt-SALDI, accompanied by prevention of the charge-up effect by the conductive Pt film. The movement of systemic insecticides in plants could be observed clearly using Pt-SALDI-IMS. The distribution and movement of components of systemic insecticides on leaves could be analyzed directly using Pt-SALDI-IMS. Additionally, changes in the properties of the chemicals with time, as an indicator of the permeability of the insecticides, could be evaluated.

  6. Nanoporous GaN-Ag composite materials prepared by metal-assisted electroless etching for direct laser desorption-ionization mass spectrometry.

    Science.gov (United States)

    Nie, Bei; Duan, Barrett K; Bohn, Paul W

    2013-07-10

    Three-dimensional nanoporous gallium nitride(PGaN) produced by metal-assisted electroless etching is chemically embedded with silver nanoparticles via electroless deposition, forming a metallized semiconductor membrane with large surface area and nanoscale metal features. A new application utilizing the unique chemical and morphological features of these composite nanostructures is described here, laser induced desorption-ionization(LDI) of biomolecules(e.g., cholesterol and nucleotides) for direct mass analysis, without use of additional organic matrix. Although PGaN itself is a poor matrix for direct LDI mass spectrometry, the combination of Ag and PGaN greatly improves ion signals relative to PGaN or Ag nanostructure surfaces alone. This behavior is attributed to the combination of strong UV absorption, enhanced surface area, and favorable thermal properties of PGaN. Importantly, Ag-PGaN is shown to facilitate the formation of Ag adduct ions in some cases, for example adenine, where adducts are not observed from either porous anodic aluminum membranes or surfaces presenting Ag nanoparticles in isolation. Nanopore-embedded Ag nanostructures serve a dual role: as cationization agents and to assist thermal desorption under UV laser irradiation. The results reported here suggest that the combination of Ag nanostructures embedded in PGaN has the capacity for high quality matrix-free LDI mass analysis.

  7. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) coupled to XAD fractionation: Method to algal organic matter characterization.

    Science.gov (United States)

    Nicolau, Rudy; Leloup, Maud; Lachassagne, Delphine; Pinault, Emilie; Feuillade-Cathalifaud, Geneviève

    2015-05-01

    This work is focused on the development of an analytical procedure for the improvement of the Organic Matter structure characterization, particularly the algal matter. Two fractions of algal organic matter from laboratory cultures of algae (Euglena gracilis) and cyanobacteria (Microcystis aeruginosa) were extracted with XAD resins. The fractions were studied using laser desorption ionization (LDI) and Matrix-Assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF). A comparison with the natural organic matter characteristics from commercial humic acids and fulvic acids extracted from Suwannee River was performed. Results show that algal and natural organic matters have unique quasi-polymeric structures. Significant repeating patterns were identified. Different fractions extracted from organic matter with common origin had common structures. Thus, 44, 114 and 169Da peaks separation for fractions from E. gracilis organic matter and 28, 58 and 100Da for M. aeruginosa ones were clearly observed. Using the developed protocol, a structural scheme and organic matter composition were obtained. The range 600-2000Da contained more architectural composition differences than the range 100-600Da, suggesting that organic matter is composed of an assembly of common small molecules. Associated to specific monomers, particular patterns were common to all samples but assembly and resulting structure were unique for each organic matter. Thus, XAD fractionation coupled to mass spectroscopy allowed determining a specific fingerprint for each organic matter.

  8. Analysis of Antiretrovirals in Single Hair Strands for Evaluation of Drug Adherence with Infrared-Matrix-Assisted Laser Desorption Electrospray Ionization Mass Spectrometry Imaging.

    Science.gov (United States)

    Rosen, Elias P; Thompson, Corbin G; Bokhart, Mark T; Prince, Heather M A; Sykes, Craig; Muddiman, David C; Kashuba, Angela D M

    2016-01-19

    Adherence to a drug regimen can be a strong predictor of health outcomes, and validated measures of adherence are necessary at all stages of therapy from drug development to prescription. Many of the existing metrics of drug adherence (e.g., self-report, pill counts, blood monitoring) have limitations, and analysis of hair strands has recently emerged as an objective alternative. Traditional methods of hair analysis based on LC-MS/MS (segmenting strands at ≥1 cm length) are not capable of preserving a temporal record of drug intake at higher resolution than approximately 1 month. Here, we evaluated the detectability of HIV antiretrovirals (ARVs) in hair from a range of drug classes using infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry imaging (MSI) with 100 μm resolution. Infrared laser desorption of hair strands was shown to penetrate into the strand cortex, allowing direct measurement by MSI without analyte extraction. Using optimized desorption conditions, a linear correlation between IR-MALDESI ion abundance and LC-MS/MS response was observed for six common ARVs with estimated limits of detection less than or equal to 1.6 ng/mg hair. The distribution of efavirenz (EFV) was then monitored in a series of hair strands collected from HIV infected, virologically suppressed patients. Because of the role hair melanin plays in accumulation of basic drugs (like most ARVs), an MSI method to quantify the melanin biomarker pyrrole-2,3,5-tricarboxylic acid (PTCA) was evaluated as a means of normalizing drug response between patients to develop broadly applicable adherence criteria.

  9. Study of bisphosphonates by matrix-assisted laser desorption/ionization mass spectrometry--influence of alkali atoms on fragmentation patterns.

    Science.gov (United States)

    Guénin, Erwann; Lecouvey, Marc; Hardouin, Julie

    2009-05-01

    1-hydroxymethylene-1,1-bisphosphonic acids (or bisphosphonates) are compounds that have interesting pharmacological applications. However, few mass spectrometric investigations have been carried out to determine their fragmentation patterns. Herein, we evaluated different matrices for the study by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) of the formation and fragmentation of the protonated, the cationized (MNa+ and MK+) and the deprotonated bisphosphonates. Some in-source fragmentations were observed both in positive and in negative ion modes. The fragmentation patterns obtained in post-source decay mode are also discussed. In contrast to previous electrospray ionization/multi-stage mass spectrometry (ESI-MSn) studies, some new fragmentation pathways were deduced and the effects of alkali ions on the fragmentation patterns were shown. The results summarized here completed the data previously recorded by ESI-MSn and could be used for the characterization of bisphosphonates as alkali complexes in biological mixtures.

  10. Reproducibility of serum protein profiling by systematic assessment using solid-phase extraction and matrix-assisted laser desorption/ionization mass spectrometry

    DEFF Research Database (Denmark)

    Callesen, Anne K; Christensen, René Depont; Madsen, Jonna S

    2008-01-01

    for serum protein profiling we investigated a range of sample preparation techniques and developed a statistical method based on repeated analyses for evaluation of protein-profiling performance of MALDI MS. Two different solid-phase extraction (SPE) methods were investigated, namely custom......Protein profiling of human serum by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) is potentially a new diagnostic tool for early detection of human diseases, including cancer. Sample preparation is a key issue in MALDI MS and the analysis of complex samples such as serum......-made microcolumns and commercially available magnetic beads. Using these two methods, nineteen different sample preparation methods for serum profiling by MALDI MS were systematically tested with regard to matrix selection, stationary phase, selectivity, and reproducibility. Microcolumns were tested with regard...

  11. Comparative study of matrix-assisted laser desorption/ionization and gas chromatography for quantitative determination of cocoa butter and cocoa butter equivalent triacylglycerol composition.

    Science.gov (United States)

    Guyon, F; Absalon, Ch; Eloy, A; Salagoity, M H; Esclapez, M; Medina, B

    2003-01-01

    The triacylglycerol (TAG) composition study of cocoa butter (CB) and cocoa butter equivalents (CBEs) has been performed by gas chromatography (GC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). These two techniques provided comparable results. The advantage of the MALDI technique was the detection of each compound comprising the triacylglycerol classes (Cn). Moreover, comparison of the data obtained by these two techniques indicated that TAG relative percentages could be obtained quantitatively with the MALDI technique. These techniques have been applied for the composition determination of CB + CBE mixtures. Encouraging results showed that it is possible to quantify an admixture containing as little as 4% of CBE.

  12. Direct Analysis of Triterpenes from High-Salt Fermented Cucumbers Using Infrared Matrix-Assisted Laser Desorption Electrospray Ionization (IR-MALDESI)

    Science.gov (United States)

    Ekelöf, Måns; McMurtrie, Erin K.; Nazari, Milad; Johanningsmeier, Suzanne D.; Muddiman, David C.

    2017-02-01

    High-salt samples present a challenge to mass spectrometry (MS) analysis, particularly when electrospray ionization (ESI) is used, requiring extensive sample preparation steps such as desalting, extraction, and purification. In this study, infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) coupled to a Q Exactive Plus mass spectrometer was used to directly analyze 50-μm thick slices of cucumber fermented and stored in 1 M sodium chloride brine. From the several hundred unique substances observed, three triterpenoid lipids produced by cucumbers, β-sitosterol, stigmasterol, and lupeol, were putatively identified based on exact mass and selected for structural analysis. The spatial distribution of the lipids were imaged, and the putative assignments were confirmed by tandem mass spectrometry performed directly on the same cucumber, demonstrating the capacity of the technique to deliver confident identifications from highly complex samples in molar concentrations of salt without the need for sample preparation.

  13. Detection of Radical Adducts with Small Molecular Weights by Matrix-Assisted Laser Desorption/Ionization with Fourier Transform Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    TIAN,Yao-Wei; SUN,Shi-Hao; XIE,Jian-Ping; ZONG,Yong-Li; NIE,Cong; GUO,Yin-Long

    2007-01-01

    As an alternative method, matrix-assisted laser desorption/ionization with Fourier transform mass spectrometry (MALDI-FTMS) has been successfully used to detect and identify free radical adducts with small molecular weights of hydroxyl and 2-cyano-2-propyl radicals trapped with 5,5-dimethylpyrroline N-oxide (DMPO). The detection and identification by MS/MS experiments using sustained offresonance irradiation collision-induced dissociation (SORI-CID) of [(DMPO+·OH-·H)+H+] (m/z 130.0868) and [DMPO+2 ·CH(CH3)2CN+H+] (m/z 250.1917) have demonstrated that MALDI-FTMS could be an effective method for detection and identification of free radical adducts. Other radical adducts have been also detected and identified. The approach of MALDI-FTMS is simple, fast, and sensitive which has potential for high-throughput analysis.

  14. Structural determination of the conjugate of human serum albumin with a mitomycin C derivative, KW-2149, by matrix-assisted laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Yasuzawa, T; Tomer, K B

    1997-01-01

    A new mitomycin C derivative, KW-2149, is known to form a covalent conjugate with human serum albumin (HSA). This conjugate exhibits 1/20 of the anticellular activity of unconjugated KW-2149. Structural studies of this conjugate were carried out using a combination of enzymatic digestion, high-performance liquid chromatography (HPLC), and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The tryptic peptide T5 (residues 21-41) was the only peptide found to be modified by KW-2149 moieties, the [(gamma-L-glutamylamino)ethyl]thio group or the (2-aminoethyl)thio group, through a disulfide bond. Although the latter peptide lost its mitomycin C moiety in the course of tryptic digestion, these data strongly suggest that KW-2149 was bound to Cys-34, the only free cysteine on HSA.

  15. An extraction method of positive blood cultures for direct identification of Candida species by Vitek MS matrix-assisted laser desorption ionization time of flight mass spectrometry.

    Science.gov (United States)

    Lavergne, Rose-Anne; Chauvin, Pamela; Valentin, Alexis; Fillaux, Judith; Roques-Malecaze, Christine; Arnaud, Sylvie; Menard, Sandie; Magnaval, Jean-François; Berry, Antoine; Cassaing, Sophie; Iriart, Xavier

    2013-08-01

    Candida spp. are an important cause of nosocomial bloodstream infections. Currently, complete identification of yeasts with conventional methods takes several days. We report here the first evaluation of an extraction method associated with the Vitek MS matrix-assisted laser desorption ionization time of flight mass spectrometry for direct identification of Candida species from positive blood cultures. We evaluated this protocol with blood cultures that were inoculated with reference and routine isolates (eight reference strains, 30 patients isolates and six mixed cultures containing two strains of different Candida species), or from patients with candidemia (28 isolates). This method performed extremely well (97% correct identification) with blood cultures of single Candida spp. and significantly reduced the time of diagnosis. Nevertheless, subculture remains indispensable to test fungal resistance and to detect mixed infections.

  16. Preparation of porous styrenics-based monolithic layers for thin layer chromatography coupled with matrix-assisted laser-desorption/ionization time-of-flight mass spectrometric detection.

    Science.gov (United States)

    Lv, Yongqin; Lin, Zhixing; Tan, Tianwei; Svec, Frantisek

    2013-11-01

    Monolithic 50 μm thin poly(4-methylstyrene-co-chloromethylstyrene-co-divinylbenzene) layers attached to 6.0 cm × 3.3 cm glass plates have been prepared, using a thermally initiated polymerization process. These layers had a well-defined porous structure with a globular morphology demonstrated with SEM images and exhibited superhydrophobic properties characterized with a water contact angle of 157°. They were then used for thin-layer chromatography of peptides and proteins fluorescently labeled with fluorescamine. The spots of individual separated compounds were visualized using UV light, and their identities were confirmed with a matrix-assisted laser desorption/ionization time of flight mass spectrometry. The presence of chloromethylstyrene units in the polymer enabled hypercrosslinking via a Friedel-Crafts alkylation reaction, and led to monoliths with much larger surface areas, which were suitable for separations of small dye molecules.

  17. Analysis of carbohydrates and glycoconjugates by matrix-assisted laser desorption/ionization mass spectrometry: An update for 2003-2004.

    Science.gov (United States)

    Harvey, David J

    2009-01-01

    This review is the third update of the original review, published in 1999, on the application of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry to the analysis of carbohydrates and glycoconjugates and brings the topic to the end of 2004. Both fundamental studies and applications are covered. The main topics include methodological developments, matrices, fragmentation of carbohydrates and applications to large polymeric carbohydrates from plants, glycans from glycoproteins and those from various glycolipids. Other topics include the use of MALDI MS to study enzymes related to carbohydrate biosynthesis and degradation, its use in industrial processes, particularly biopharmaceuticals and its use to monitor products of chemical synthesis where glycodendrimers and carbohydrate-protein complexes are highlighted.

  18. Feasibility of matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) networking in university hospitals in Brussels.

    Science.gov (United States)

    Martiny, D; Cremagnani, P; Gaillard, A; Miendje Deyi, V Y; Mascart, G; Ebraert, A; Attalibi, S; Dediste, A; Vandenberg, O

    2014-05-01

    The mutualisation of analytical platforms might be used to address rising healthcare costs. Our study aimed to evaluate the feasibility of networking a unique matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) system for common use in several university hospitals in Brussels, Belgium. During a one-month period, 1,055 successive bacterial isolates from the Brugmann University Hospital were identified on-site using conventional techniques; these same isolates were also identified using a MALDI-TOF MS system at the Porte de Hal Laboratory by sending target plates and identification projects via transportation and the INFECTIO_MALDI software (Infopartner, Nancy, France), respectively. The occurrence of transmission problems (helpdesk to manage potential connectivity problems.

  19. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry as a tool for fast identification of protein binders in color layers of paintings.

    Science.gov (United States)

    Hynek, Radovan; Kuckova, Stepanka; Hradilova, Janka; Kodicek, Milan

    2004-01-01

    Identification of materials in color layers of paintings is necessary for correct decisions concerning restoration procedures as well as proving the authenticity of the painting. The proteins are usually important components of the painting layers. In this paper it has been demonstrated that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) can be used for fast and reliable identification of proteins in color layers even in old, highly aged matrices. The digestion can be easily performed directly on silica wafers which are routinely used for infrared analysis. The amount of material necessary for such an analysis is extremely small. Peptide mass mapping using digestion with trypsin followed by MALDI-TOFMS and identification of the protein was successfully used for determination of the binder from a painting of the 19th century.

  20. Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Analysis of Gram-Positive, Catalase-Negative Cocci Not Belonging to the Streptococcus or Enterococcus Genus and Benefits of Database Extension

    DEFF Research Database (Denmark)

    Christensen, Jens Jørgen; Dargis, Rimtas; Hammer, Monja;

    2012-01-01

    Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry with a Bruker Daltonics microflex LT system was applied to 90 well-characterized catalase-negative, Gram-positive cocci not belonging to the streptococci or enterococci. Biotyper version 2.0.43.1 software was...

  1. Classification of wheat varieties: Use of two-dimensional gel electrophoresis for varieties that can not be classified by matrix assisted laser desorption/ionization-time of flight-mass spectrometry and an artificial neural network

    DEFF Research Database (Denmark)

    Jacobsen, Susanne; Nesic, Ljiljana; Petersen, Marianne Kjerstine;

    2001-01-01

    Analyzing a gliadin extract by matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI- TOF-MS) combined with an artificial neural network (ANN) is a suitable method for identification of wheat varieties. However, the ANN can not distinguish between all different wheat...

  2. Bacteriophage cell lysis of Shiga toxin-producing Escherichia coli for top-down proteomic identification of Shiga toxin 1 & 2 using matrix-assisted laser desorption/ionization tandem time-of-light mass spectrometry

    Science.gov (United States)

    RATIONALE: Analysis of bacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) often relies upon sample preparation methods that result in cell lysis, e.g. bead-beating. However, Shiga toxin-producing Escherichia coli (STEC) can undergo bacteriophage...

  3. On-Tissue Derivatization via Electrospray Deposition for Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging of Endogenous Fatty Acids in Rat Brain Tissues.

    Science.gov (United States)

    Wu, Qian; Comi, Troy J; Li, Bin; Rubakhin, Stanislav S; Sweedler, Jonathan V

    2016-06-07

    Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) is used for the multiplex detection and characterization of diverse analytes over a wide mass range directly from tissues. However, analyte coverage with MALDI MSI is typically limited to the more abundant compounds, which have m/z values that are distinct from MALDI matrix-related ions. On-tissue analyte derivatization addresses these issues by selectively tagging functional groups specific to a class of analytes, while simultaneously changing their molecular masses and improving their desorption and ionization efficiency. We evaluated electrospray deposition of liquid-phase derivatization agents as a means of on-tissue analyte derivatization using 2-picolylamine; we were able to detect a range of endogenous fatty acids with MALDI MSI. When compared with airbrush application, electrospray led to a 3-fold improvement in detection limits and decreased analyte delocalization. Six fatty acids were detected and visualized from rat cerebrum tissue using a MALDI MSI instrument operating in positive mode. MALDI MSI of the hippocampal area allowed targeted fatty acid analysis of the dentate gyrus granule cell layer and the CA1 pyramidal layer with a 20-μm pixel width, without degrading the localization of other lipids during liquid-phase analyte derivatization.

  4. Sequencing of phosphopeptides sulfonated by 4-sulfophenyl isothiocyanate using post-source decay matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    TANG Jianguang; WANG Yongjun; ZHANG Hainan; WANG Chunyu; HU Zhiping; XUE Zhigang; XIA Kun; SHI Xiaoliu

    2006-01-01

    Phosphorylation/dephosphorylation is probably the most common and important reversible post-translational modificaion of proteins. Analyzing the functional effects of phosphorylation is helpful for understanding the biological functions of proteins. Identification of the phosphorylation sites of phosphorylated protein is a prerequisite for research on phosphorylation. In this work, an effective and simple method of identification of protein phosphorylation sites has been developed. Phosphopeptides were selectively enriched with immobilized metal affinity chromatography (IMAC) and subsequently chemically modified by 4-sulfophenyl isothiocyanate, and then the chemically modified phosphopeptides were sequenced with post-source decay (PSD) matrix-assisted laser desorption/ionization (MALDI)time-of-flight mass spectrometry for detecting phosphorylation sites. The charge of derivatization by 4-sulfophenyl isothiocyanate introduces a negative sulfonic acid group at the N-terminus of a peptide, and enables the selective detection of only a single series of C-terminal y-type ions. This chemically assisted method greatly simplifies the extremely complex pattern of PSD fragment ions and makes the PSD spectra more easier to be interpreted. The phosphorylation sites of a synthesized model phosphopeptide and human c-myc protein have been successfully identified by this method.Phosphorylation/dephosphorylation is probably the most common and important reversible post-translational modificaion of proteins. Analyzing the functional effects of phosphorylation is helpful for understanding the biological functions of proteins. Identification of the phosphorylation sites of phosphorylated protein is a prerequisite for research on phosphorylation. In this work, an effective and simple method of identification of protein phosphorylation sites has been developed. Phosphopeptides were selectively enriched with immobilized metal affinity chromatography (IMAC) and subsequently chemically

  5. Carbon Dots and 9AA as a Binary Matrix for the Detection of Small Molecules by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

    Science.gov (United States)

    Chen, Yongli; Gao, Dan; Bai, Hangrui; Liu, Hongxia; Lin, Shuo; Jiang, Yuyang

    2016-07-01

    Application of matrix-assisted laser-desorption/ionization mass spectrometry (MALDI MS) to analyze small molecules have some limitations, due to the inhomogeneous analyte/matrix co-crystallization and interference of matrix-related peaks in low m/z region. In this work, carbon dots (CDs) were for the first time applied as a binary matrix with 9-Aminoacridine (9AA) in MALDI MS for small molecules analysis. By 9AA/CDs assisted desorption/ionization (D/I) process, a wide range of small molecules, including nucleosides, amino acids, oligosaccharides, peptides, and anticancer drugs with a higher sensitivity were demonstrated in the positive ion mode. A detection limit down to 5 fmol was achieved for cytidine. 9AA/CDs matrix also exhibited excellent reproducibility compared with 9AA matrix. Moreover, by exploring the ionization mechanism of the matrix, the influence factors might be attributed to the four parts: (1) the strong UV absorption of 9AA/CDs due to their π-conjugated network; (2) the carboxyl groups modified on the CDs surface act as protonation sites for proton transfer in positive ion mode; (3) the thin layer crystal of 9AA/CDs could reach a high surface temperature more easily and lower transfer energy for LDI MS; (4) CDs could serve as a matrix additive to suppress 9AA ionization. Furthermore, this matrix was allowed for the analysis of glucose as well as nucleosides in human urine, and the level of cytidine was quantified with a linear range of 0.05-5 mM (R2 > 0.99). Therefore, the 9AA/CDs matrix was proven to be an effective MALDI matrix for the analysis of small molecules with improved sensitivity and reproducibility. This work provides an alternative solution for small molecules detection that can be further used in complex samples analysis.

  6. Carbon Dots and 9AA as a Binary Matrix for the Detection of Small Molecules by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.

    Science.gov (United States)

    Chen, Yongli; Gao, Dan; Bai, Hangrui; Liu, Hongxia; Lin, Shuo; Jiang, Yuyang

    2016-07-01

    Application of matrix-assisted laser-desorption/ionization mass spectrometry (MALDI MS) to analyze small molecules have some limitations, due to the inhomogeneous analyte/matrix co-crystallization and interference of matrix-related peaks in low m/z region. In this work, carbon dots (CDs) were for the first time applied as a binary matrix with 9-Aminoacridine (9AA) in MALDI MS for small molecules analysis. By 9AA/CDs assisted desorption/ionization (D/I) process, a wide range of small molecules, including nucleosides, amino acids, oligosaccharides, peptides, and anticancer drugs with a higher sensitivity were demonstrated in the positive ion mode. A detection limit down to 5 fmol was achieved for cytidine. 9AA/CDs matrix also exhibited excellent reproducibility compared with 9AA matrix. Moreover, by exploring the ionization mechanism of the matrix, the influence factors might be attributed to the four parts: (1) the strong UV absorption of 9AA/CDs due to their π-conjugated network; (2) the carboxyl groups modified on the CDs surface act as protonation sites for proton transfer in positive ion mode; (3) the thin layer crystal of 9AA/CDs could reach a high surface temperature more easily and lower transfer energy for LDI MS; (4) CDs could serve as a matrix additive to suppress 9AA ionization. Furthermore, this matrix was allowed for the analysis of glucose as well as nucleosides in human urine, and the level of cytidine was quantified with a linear range of 0.05-5 mM (R(2) > 0.99). Therefore, the 9AA/CDs matrix was proven to be an effective MALDI matrix for the analysis of small molecules with improved sensitivity and reproducibility. This work provides an alternative solution for small molecules detection that can be further used in complex samples analysis. Graphical Abstract ᅟ.

  7. Infrared laser desorption/ionization on silicon.

    Science.gov (United States)

    Bhattacharya, Sucharita H; Raiford, Timothy J; Murray, Kermit K

    2002-05-01

    Laser desorption/ionization from a single-crystal silicon surface was performed using a laser operating in the 3-microm region of the mid-infrared. Analyte molecules up to 6 kDa were ionized with no added matrix. As with ultraviolet desorption/ionization from porous silicon (DIOS), IR laser desorption from silicon does not produce matrix ions that can interfere with analysis of low-mass analytes. However, in contrast to UV DIOS, silicon porosity or roughness is not required for ionization using an IR laser. Mass spectra were obtained in the wavelength range between 2.8 and 3.5 microm, which is consistent with energy absorption by a hydrogen-bonded OH group. A mechanism based on desorption of adsorbed solvent molecules is postulated.

  8. Efficiency of Gas-Phase Ion Formation in Matrix-Assisted Laser Desorption Ionization with 2,5-Dihydroxybenzoic Acid as Matrix

    Energy Technology Data Exchange (ETDEWEB)

    Park, Kyung Man; Ahn, Sung Hee; Bae, Yong Jin; Kim, Myung Soo [Seoul National Univ., Seoul (Korea, Republic of)

    2013-03-15

    Numbers of matrix- and analyte-derived ions and their sum in matrix-assisted laser desorption ionization (MALDI) of a peptide were measured using 2,5-dihydroxybenzoic acid (DHB) as matrix. As for MALDI with α-cyano-4-hydroxy cinnamic acid as matrix, the sum was independent of the peptide concentration in the solid sample, or was the same as that of pure DHB. This suggested that the matrix ion was the primary ion and that the peptide ion was generated by matrix-to-peptide proton transfer. Experimental ionization efficiencies of 10{sup -5}-10{sup -4} for peptides and 10{sup -8}-10{sup -7} for matrices are far smaller than 10.3-10.1 for peptides and 10{sup -5}-10{sup -3} for matrices speculated by Hillenkamp and Karas. Number of gas-phase ions generated by MALDI was unaffected by laser wavelength or pulse energy. This suggests that the main role of photo-absorption in MALDI is not in generating ions via a multi-photon process but in ablating materials in a solid sample to the gas phase.

  9. Generation of highly charged peptide and protein ions by atmospheric pressure matrix-assisted infrared laser desorption/ionization ion trap mass spectrometry.

    Science.gov (United States)

    König, Simone; Kollas, Oliver; Dreisewerd, Klaus

    2007-07-15

    We show that highly charged ions can be generated if a pulsed infrared laser and a glycerol matrix are employed for atmospheric pressure matrix-assisted laser desorption/ionization mass spectrometry with a quadrupole ion trap. Already for small peptides like bradykinin, doubly protonated ions form the most abundant analyte signal in the mass spectra. The center of the charge-state distribution increases with the size of the analyte. For example, insulin is detected with a most abundant ion signal corresponding to a charge state of four, whereas for cytochrome c, the 10 times protonated ion species produces the most intense signal. Myoglobin is observed with up to 13 charges. The high m/z ratios allow us to use the Paul trap for the detection of MALDI-generated protein ions that are, owing to their high molecular weight, not amenable in their singly protonated charge state. Formation of multiple charges critically depends on the addition of diluted acid to the analyte-matrix solution. Tandem mass spectra generated by collision-induced dissociation of doubly charged peptides are also presented. The findings allow speculations about the involvement of electrospray ionization processes in these MALDI experiments.

  10. ARTICLES: Influence Factors on Particle Growth for On-line Aerosol Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry

    Science.gov (United States)

    Xia, Wei-wei; Ti, Ru-fang; Zhang, Zi-Iiang; Zheng, Hai-yang; Fang, Li

    2010-06-01

    An evaporation/condensation flow cell was developed and interfaced with the matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometer for on-line bioaerosol detection and characterization, which allows matrix addition by condensation onto the laboratory-generated bioaerosol particles. The final coated particle exiting from the condenser is then introduced into the aerodynamic particle sizer spectrometer or home-built aerosol laser time-of-flight mass spectrometer, and its aerodynamic size directly effects on the matrix-to-analyte molar ratio, which is very important for MALDI technique. In order to observe the protonated analyte molecular ion, and then determine the classification of biological aerosols, the matrix-to-analyte molar ratio must be appropriate. Four experimental parameters, including the temperature of the heated reservoir, the initial particle size, its number concentration, and the matrix material, were tested experimentally to analyze their influences on the final particle size. This technique represents an on-line system of detection that has the potential to provide rapid and reliable identification of airborne biological aerosols.

  11. Advanced stored waveform inverse Fourier transform technique for a matrix-assisted laser desorption/ionization quadrupole ion trap mass spectrometer.

    Science.gov (United States)

    Doroshenko, V M; Cotter, R J

    1996-01-01

    The stored waveform inverse Fourier transform (SWIFT) technique is used for broadband excitation of ions in an ion-trap mass spectrometer to perform mass-selective accumulation, isolation, and fragmentation of peptide ions formed by matrix-assisted laser desorption/ionization. Unit mass resolution is achieved for isolation of ions in the range of m/z up to 1300 using a two-step isolation technique with stretched-in-time narrow band SWIFT pulses at the second stage. The effect of 'stretched-in-time' waveforms is similar to that observed previously for mass-scan-rate reduction. The asymmetry phenomenon resulting from the stretched ion-trap electrode geometry is observed during application of normal and time-reversed waveforms and is similar to the asymmetry effects observed for forward and reverse mass scans in the resonance ejection mode. Mass-selective accumulation of ions from multiple laser shots was accomplished using a method described earlier that involves increasing the trapping voltage during ion introduction for more efficient trapping of ions.

  12. Decision peptide-driven: a free software tool for accurate protein quantification using gel electrophoresis and matrix assisted laser desorption ionization time of flight mass spectrometry.

    Science.gov (United States)

    Santos, Hugo M; Reboiro-Jato, Miguel; Glez-Peña, Daniel; Nunes-Miranda, J D; Fdez-Riverola, Florentino; Carvallo, R; Capelo, J L

    2010-09-15

    The decision peptide-driven tool implements a software application for assisting the user in a protocol for accurate protein quantification based on the following steps: (1) protein separation through gel electrophoresis; (2) in-gel protein digestion; (3) direct and inverse (18)O-labeling and (4) matrix assisted laser desorption ionization time of flight mass spectrometry, MALDI analysis. The DPD software compares the MALDI results of the direct and inverse (18)O-labeling experiments and quickly identifies those peptides with paralleled loses in different sets of a typical proteomic workflow. Those peptides are used for subsequent accurate protein quantification. The interpretation of the MALDI data from direct and inverse labeling experiments is time-consuming requiring a significant amount of time to do all comparisons manually. The DPD software shortens and simplifies the searching of the peptides that must be used for quantification from a week to just some minutes. To do so, it takes as input several MALDI spectra and aids the researcher in an automatic mode (i) to compare data from direct and inverse (18)O-labeling experiments, calculating the corresponding ratios to determine those peptides with paralleled losses throughout different sets of experiments; and (ii) allow to use those peptides as internal standards for subsequent accurate protein quantification using (18)O-labeling. In this work the DPD software is presented and explained with the quantification of protein carbonic anhydrase.

  13. Direct identification of trypanosomatids by matrix-assisted laser desorption ionization-time of flight mass spectrometry (DIT MALDI-TOF MS).

    Science.gov (United States)

    Avila, C C; Almeida, F G; Palmisano, G

    2016-08-01

    Accurate and rapid determination of trypanosomatids is essential in epidemiological surveillance and therapeutic studies. Matrix-assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF MS) has been shown to be a useful and powerful technique to identify bacteria, fungi, metazoa and human intact cells with applications in clinical settings. Here, we developed and optimized a MALDI-TOF MS method to profile trypanosomatids. trypanosomatid cells were deposited on a MALDI target plate followed by addition of matrix solution. The plate was then subjected to MALDI-TOF MS measurement to create reference mass spectra library and unknown samples were identified by pattern matching using the BioTyper software tool. Several m/z peaks reproducibly and uniquely identified trypanosomatids species showing the potentials of direct identification of trypanosomatids by MALDI-TOF MS. Moreover, this method discriminated different life stages of Trypanosoma cruzi, epimastigote and bloodstream trypomastigote and Trypanosoma brucei, procyclic and bloodstream. T. cruzi Discrete Typing Units (DTUs) were also discriminated in three clades. However, it was not possible to achieve enough resolution and software-assisted identification at the strain level. Overall, this study shows the importance of MALDI-TOF MS for the direct identification of trypanosomatids and opens new avenues for mass spectrometry-based detection of parasites in biofluids. Copyright © 2016 John Wiley & Sons, Ltd.

  14. Global optimization of the infrared matrix-assisted laser desorption electrospray ionization (IR MALDESI) source for mass spectrometry using statistical design of experiments.

    Science.gov (United States)

    Barry, Jeremy A; Muddiman, David C

    2011-12-15

    Design of experiments (DOE) is a systematic and cost-effective approach to system optimization by which the effects of multiple parameters and parameter interactions on a given response can be measured in few experiments. Herein, we describe the use of statistical DOE to improve a few of the analytical figures of merit of the infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) source for mass spectrometry. In a typical experiment, bovine cytochrome c was ionized via electrospray, and equine cytochrome c was desorbed and ionized by IR-MALDESI such that the ratio of equine:bovine was used as a measure of the ionization efficiency of IR-MALDESI. This response was used to rank the importance of seven source parameters including flow rate, laser fluence, laser repetition rate, ESI emitter to mass spectrometer inlet distance, sample stage height, sample plate voltage, and the sample to mass spectrometer inlet distance. A screening fractional factorial DOE was conducted to designate which of the seven parameters induced the greatest amount of change in the response. These important parameters (flow rate, stage height, sample to mass spectrometer inlet distance, and laser fluence) were then studied at higher resolution using a full factorial DOE to obtain the globally optimized combination of parameter settings. The optimum combination of settings was then compared with our previously determined settings to quantify the degree of improvement in detection limit. The limit of detection for the optimized conditions was approximately 10 attomoles compared with 100 femtomoles for the previous settings, which corresponds to a four orders of magnitude improvement in the detection limit of equine cytochrome c.

  15. A method to detect metal-drug complexes and their interactions with pathogenic bacteria via graphene nanosheet assist laser desorption/ionization mass spectrometry and biosensors

    Energy Technology Data Exchange (ETDEWEB)

    Abdelhamid, Hani Nasser [Department of Chemistry, National Sun Yat-Sen University, Kaohsiung, 804, Taiwan (China); Wu, Hui-Fen, E-mail: hwu@faculty.nsysu.edu.tw [Department of Chemistry, National Sun Yat-Sen University, Kaohsiung, 804, Taiwan (China); School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, 800, Taiwan (China); Center for Nanoscience and Nanotechnology, National Sun Yat-Sen University, Kaohsiung, 804, Taiwan (China); Doctoral Degree Program in Marine Biotechnology, National Sun Yat-Sen University, Kaohsiung, 804, Taiwan (China)

    2012-11-02

    Highlights: Black-Right-Pointing-Pointer Probe transition metals-complexes based on noncovalent functionalized graphene for MALDI-MS. Black-Right-Pointing-Pointer Study interaction of transition metals complexes with pathogenic bacteria. Black-Right-Pointing-Pointer Propose a new biosensor for two pathogenic bacteria. - Abstract: A new method was proposed to probe the interactions between transition metals of Fe(II), Fe(III), Cu(II) with a non steroidal anti-inflammatory drug (NSAID), flufenamic acid (FF) using graphene as a matrix for Graphene assisted laser desorption ionization mass spectrometry (GALDI-MS). Metal-drug complexation was confirmed via UV absorption spectroscopy, fluorescence spectroscopy, pH meter, and change in solution conductivity. The optimal molar ratios for these complexation interactions are stoichiometry 1:2 in both Cu(II) and Fe(II) complexes, and 1:3 in Fe(III) complexes at physiological pH (7.4). Metal complexation of the drug could enhance fluorescence for 20 fold which is due to the charge transfer reaction or increase rigidity of the drug. The main interaction between graphene and flufenamic acid is the {Pi}-{Pi} interaction which allows us to probe the metal-drug complexation. The GALDI-MS could sensitively detect the drug at m/z 281.0 Da (protonated molecule) with detection limit 2.5 pmol (1.0 {mu}M) and complexation at m/z 661.0, 654.0 and 933.0 Da corresponding to [Cu(II)(FF){sub 2}(H{sub 2}O){sub 2} + H]{sup +}, [Fe(II)(FF){sub 2}(H{sub 2}O){sub 2} + H]{sup +} and [Fe(III) (FF){sub 3}(H{sub 2}O){sub 2} + H]{sup +}, respectively (with limit of detection (LOD) 2.0 pmol (10.0 {mu}M). Matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) spectra show change in the protein profile of intact pathogenic bacteria (Pseudomonas aeroginosa, Staphylococcus aureus). The change in the ionization ability (mainly proton affinity) of pathogenic bacteria may be due to the interactions between the bacteria with the drug (or its

  16. Quantum theory of laser-stimulated desorption

    Science.gov (United States)

    Slutsky, M. S.; George, T. F.

    1978-01-01

    A quantum theory of laser-stimulated desorption (LSDE) is presented and critically analyzed. It is shown how LSDE depends on laser-pulse characteristics and surface-lattice dynamics. Predictions of the theory for a Debye model of the lattice dynamics are compared to recent experimental results.

  17. Sample-first preparation: a method for surface-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of cyclic oligosaccharides.

    Science.gov (United States)

    Wu, Hsin-Pin; Su, Chih-Lin; Chang, Hui-Chiu; Tseng, Wei-Lung

    2007-08-15

    A new sample preparation method for the analysis of cyclic oligosaccharides in surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) is presented. We call this new technique "sample first method", in which a sample is deposited first and then bare gold nanoparticles (AuNPs), which serve as the SALDI matrixes, are added to the top of the sample layer. The use of the sample first method offers significant advantages for improving shot-to-shot reproducibility, enhancing the ionization efficiency of the analyte, and reducing sample preparation time as compared to the dried-droplet method, wherein samples and bare AuNPs are mixed and dried together. The relative standard deviation (RSD) values of the signal intensity as calculated from 65 sample spots was 25% when the sample first methods were applied to the analysis of beta-cyclodextrin. The results were more homogeneous as compared to the outcome using dried-droplet preparation of AuNPs (RSD=66%) and 2,5-dihydroxybenzoic acid (RSD=209%). We also found out that the optimal concentration of AuNP for ionization efficiency is 7.4 nM (4.52x10(12) particles/mL) while the lowest detectable concentration of cyclic oligosaccharides through this approach is 0.25 microM. Except for the cyclic oligosaccharide, the proposed method was also applied to the analyses of other biological samples, including neutral carbohydrate and steroid, aminothiols, and peptides as well as proteins.

  18. Discrimination of different species from the genus Drosophila by intact protein profiling using matrix-assisted laser desorption ionization mass spectrometry

    Directory of Open Access Journals (Sweden)

    Gröger-Arndt Helke

    2010-04-01

    Full Text Available Abstract Background The use of molecular biology-based methods for species identification and establishing phylogenetic relationships has supplanted traditional methods relying on morphological characteristics. While PCR-based methods are now the commonly accepted gold standards for these types of analysis, relatively high costs, time-consuming assay development or the need for a priori information about species-specific sequences constitute major limitations. In the present study, we explored the possibility to differentiate between 13 different species from the genus Drosophila via a molecular proteomic approach. Results After establishing a simple protein extraction procedure and performing matrix-assisted laser desorption/ionization (MALDI mass spectrometry (MS with intact proteins and peptides, we could show that most of the species investigated reproducibly yielded mass spectra that were adequate for species classification. Furthermore, a dendrogram generated by cluster analysis of total protein patterns agrees reasonably well with established phylogenetic relationships. Conclusion Considering the intra- and interspecies similarities and differences between spectra obtained for specimens of closely related Drosophila species, we estimate that species typing of insects and possibly other multicellular organisms by intact protein profiling (IPP can be established successfully for species that diverged from a common ancestor about 3 million years ago.

  19. Study of Phospholipids in Single Cells Using an Integrated Microfluidic Device Combined with Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.

    Science.gov (United States)

    Xie, Weiyi; Gao, Dan; Jin, Feng; Jiang, Yuyang; Liu, Hongxia

    2015-07-21

    Single-cell trapping and high-throughput mass spectrometry analysis remain challenging now. Current technologies for single-cell analysis have several limitations, such as throughput, space resolution, and multicomponent analysis. In this study, we demonstrate, for the first time, the combination of microfluidic chip and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) for high-throughput and automatic single-cell phospholipids analysis. A microwell-array-based microfluidic chip was designed and fabricated for cell array formation on an indium tin oxide (ITO)-coated glass slide. Mass spectrometry imaging measurement with 25 μm pixel size was performed with a MALDI ion source. Eight phospholipids in a single A549 cell were detected, and their structures were further identified by MS/MS spectra. Selected ion images were generated with a bin width of Δm/z ± 0.005. The selected ion images and optical images of the cell array showed excellent correlation, and mass spectrometry information on phospholipids from 1-3 cells was extracted automatically by selecting pixels with the same fixed interval between microwells on the chip. The measurement and data extraction could be processed in several minutes to achieve a high-throughput analysis. Through the optimization of different microwell sizes and different matrices, this method showed potential for the analysis of other metabolites or metabolic changes at the single-cell level.

  20. Examination of the translocation of sulfonylurea herbicides in sunflower plants by matrix-assisted laser desorption/ionisation mass spectrometry imaging.

    Science.gov (United States)

    Anderson, David M G; Carolan, Vikki A; Crosland, Susan; Sharples, Kate R; Clench, Malcolm R

    2010-11-30

    Pesticides are widely used in agriculture to control weeds, pests and diseases. Successful control is dependent on the compound reaching the target site within the organism after spray or soil application. Conventional methods for determining uptake and movement of herbicides and pesticides include autoradiography, liquid scintillation and chromatographic techniques such as high-performance liquid chromatography (HPLC). Autoradiography using radiolabelled compounds provides the best indication of a compound's movement within the plant system. Autoradiography is an established technique but it relies on the synthesis of radiolabelled compounds. The distribution of four sulfonylurea herbicides in sunflower plants has been studied 24  h after foliar application. The use of matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) images of protonated molecules and fragment ions (resulting from fragmentation at the urea bond within the sulfonylurea herbicides) has provided evidence for translocation above and below the application point. The translocation of nicosulfuron and azoxystrobin within the same plant system has also been demonstrated following their application to the plant stem. This study provides evidence that MALDI-MSI has great potential as an analytical technique to detect and assess the foliar, root and stem uptake of agrochemicals, and to reveal their distribution through the plant once absorbed and translocated.

  1. Probing chain-end functionalization reactions in living anionic polymerization via matrix-assisted laser desorption ionization time-of-flight mass spectrometry

    Science.gov (United States)

    Arnould, Mark A.; Polce, Michael J.; Quirk, Roderic P.; Wesdemiotis, Chrys

    2004-11-01

    Matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) is applied to examine the products arising upon the preparation of chain-end functional polymers via living anionic polymerization techniques. Both post-polymerization functionalizations as well as the use of functionalized initiators are investigated. MALDI-TOF MS is shown to be a sensitive probe for the qualitative analysis of the major and minor oligomers from novel functionalization reactions whose mechanisms are not yet well established. The method is particularly valuable for the identification of the end groups of the minor, and often unexpected, distributions that may be undetectable by other analytical means. Complete characterization of all oligomers generated during functionalization reactions provides an essential tool to the synthetic chemist for understanding the corresponding mechanisms. This insight is necessary for selecting alternative routes or making modifications to the reaction conditions. It is demonstrated that MALDI-TOF MS can convey quantitative information about the yields of the chain-end groups introduced during functionalization. From the cases presented it is evident that post-polymerization reactions allow for better control of chain-end functionality and molecular weight than functionalization with the limited number of currently available protected functionalized initiators.

  2. Solvent selection for matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of synthetic polymers employing solubility parameters.

    Science.gov (United States)

    Brandt, Heike; Ehmann, Thomas; Otto, Matthias

    2010-08-30

    The principle relating to the selection of a proper matrix, cationization reagent, and solvent for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) of synthetic polymers is still a topic of research. In this work we focused on the selection of a suitable MALDI solvent. Polystyrene PS7600 and poly(ethylene glycol) PEG4820 were analyzed by MALDI-TOF MS using various solvents which were selected based on the Hansen solubility parameter system. For polystyrene (PS), dithranol was used as the matrix and silver trifluoroacetate as the cationization reagent whereas, for poly(ethylene glycol) (PEG), the combination of 2,5-dihydroxybenzoic acid and sodium trifluoroacetate was used for all experiments. When employing solvents which dissolve PS and PEG, reliable MALDI mass spectra were obtained while samples in non-solvents (solvents which are not able to dissolve the polymer) failed to provide spectra. It seems that the solubility of the matrix and the cationization reagent are less important than the polymer solubility.

  3. Detection of methicillin-resistant Staphylococcus aureus using phage amplification combined with matrix-assisted laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Rees, Jon C; Barr, John R

    2017-02-01

    Antibiotic resistance continues to contribute significantly to morbidity and mortality across the world. Developing new tests for antibiotic-resistant bacteria is a core action to combat resistant infections. We describe a method that uses phage amplification detection (PAD) combined with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to rapidly identify Staphylococcus aureus and determine phenotypic susceptibility to cefoxitin. Samples tested for S. aureus are incubated together with bacteriophage in the presence and absence of cefoxitin and subjected to rapid trypsin digestion followed by MALDI-MS analysis. Tryptic peptides derived from amplified phage proteins can be detected by MALDI-MS, as validated by time-of-flight (TOF)/TOF analysis of each peptide combined with database searching. Methicillin-resistant S. aureus show significant phage amplification in the presence of cefoxitin, while methicillin-sensitive S. aureus show no phage amplification relative to a no-antibiotic control. We also show that PAD methodology can be implemented on an FDA-approved commercial MALDI-MS bacterial identification system to identify S. aureus and determine antibiotic susceptibility. The novelty of this assay includes the use of phage-derived tryptic peptides as detected by MALDI-MS to monitor the results of PAD on an instrument common to many modern microbiology laboratories.

  4. Visualization of phosphatidylcholine, lysophosphatidylcholine and sphingomyelin in mouse tongue body by matrix-assisted laser desorption/ionization imaging mass spectrometry.

    Science.gov (United States)

    Enomoto, Hirofumi; Sugiura, Yuki; Setou, Mitsutoshi; Zaima, Nobuhiro

    2011-06-01

    The mammalian tongue is one of the most important organs during food uptake because it is helpful for mastication and swallowing. In addition, taste receptors are present on the surface of the tongue. Lipids are the second most abundant biomolecules after water in the tongue. Lipids such as phosphatidylcholine (PC), lysophosphatidylcholine (LPC) and sphingomyelin (SM) are considered to play fundamental roles in the mediation of cell signaling. Imaging mass spectrometry (IMS) is powerful tool for determining and visualizing the distribution of lipids across sections of dissected tissue. In this study, we identified and visualized the PC, LPC, and SM species in a mouse tongue body section with matrix-assisted laser desorption/ionization (MALDI)-IMS. The ion image constructed from the peaks revealed that docosahexaenoic acid (DHA)-containing PC, LPC, linoleic acid-containing PC and SM (d18:1/16:0), and oleic acid-containing PC were mainly distributed in muscle, connective tissue, stratified epithelium, and the peripheral nerve, respectively. Furthermore, the distribution of SM (d18:1/16:0) corresponded to the distribution of nerve tissue relating to taste in the stratified epithelium. This study represents the first visualization of PC, LPC and SM localization in the mouse tongue body.

  5. Increased detection rate of melamine-containing calcium urolithiasis by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry technique in clinical practice.

    Science.gov (United States)

    Wu, Chia-Fang; Liu, Chia-Chu; Chou, Yii-Her; Shiea, Jentaie; Shen, Jung-Tsung; Wang, Shiun-Shiuan; Wu, Ming-Tsang

    2014-04-20

    Background: Studies have shown that melamine may be associated with urolithiasis. A more sensitive method is needed to analyze melamine in urinary stones to identify potential causes of urolithiasis.Methods: Here we compare the analytical methods of detecting melamine in urinary stones by Fourier transform infrared (FTIR) spectroscopy and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS) in the laboratory and clinic. First, we established the melamine detection limit in melamine cyanurate standard by the methods of FTIR spectrophotometer and MALDI-TOF MS. Subsequently, we applied these two methods to 54 adult patients with upper urinary tract calcium urolithiasis.Results: We found that the detection limit of melamine in melamine cyanurate standard by MALD-TOF MS was~10,000-fold more sensitive than FTIR.We applied both instruments to 54 stone specimens from 54 calcium urolithias is patients. In those without distinctive melamine pattern in the FTIR spectra,melamine could be detected by MALD-TOF MS in an additional 12 out of 42 subjects' stone specimens (28.6%). Compared to MALD-TOF MS negative subjects (n = 30), those positive subjects (n = 12) excreted significantly higher urinary melamine levels (P melamine in melamine-containing kidney stones

  6. Investigations of the lysophospholipid composition of human neutrophils under different stimulation conditions by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Directory of Open Access Journals (Sweden)

    JURGEN ARNHOLD

    2002-03-01

    Full Text Available Matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS is usually used for the analyses of proteins, carbohydrates and oligonucleotides. In spite of the number of advantages that MALDI-TOF MS exhibits for lipid analysis, this method has not often been applied in this field. In this paper we have extended our previous studies on the suitability of MALDI-TOF MS for the investigation of changes in the content of lipid-derived second messengers in organic extracts of human neutrophils. Qualitative differences in the lysophospholipid composition in organic extracts of the human neutrophils under different stimulation conditions could be easily observed by MALDI-TOF MS. Although there are still some methodological problems to be solved before this method can be routinely applied for the quantification of different lipid classes in complex biological mixtures (such as organic extracts of human neutrophils it is shown here that MALDI-TOF MS possesses the capability to be used as a simple screening method for the investigation of the content of lipid-derived second messengers and of signalling pathways in cells.

  7. Matrix-assisted laser desorption ionization-time of flight mass spectrometry: a fundamental shift in the routine practice of clinical microbiology.

    Science.gov (United States)

    Clark, Andrew E; Kaleta, Erin J; Arora, Amit; Wolk, Donna M

    2013-07-01

    Within the past decade, clinical microbiology laboratories experienced revolutionary changes in the way in which microorganisms are identified, moving away from slow, traditional microbial identification algorithms toward rapid molecular methods and mass spectrometry (MS). Historically, MS was clinically utilized as a high-complexity method adapted for protein-centered analysis of samples in chemistry and hematology laboratories. Today, matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) MS is adapted for use in microbiology laboratories, where it serves as a paradigm-shifting, rapid, and robust method for accurate microbial identification. Multiple instrument platforms, marketed by well-established manufacturers, are beginning to displace automated phenotypic identification instruments and in some cases genetic sequence-based identification practices. This review summarizes the current position of MALDI-TOF MS in clinical research and in diagnostic clinical microbiology laboratories and serves as a primer to examine the "nuts and bolts" of MALDI-TOF MS, highlighting research associated with sample preparation, spectral analysis, and accuracy. Currently available MALDI-TOF MS hardware and software platforms that support the use of MALDI-TOF with direct and precultured specimens and integration of the technology into the laboratory workflow are also discussed. Finally, this review closes with a prospective view of the future of MALDI-TOF MS in the clinical microbiology laboratory to accelerate diagnosis and microbial identification to improve patient care.

  8. Evaluation of synthase and hemisynthase activities of glucosamine-6-phosphate synthase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Gaucher-Wieczorek, Florence; Guérineau, Vincent; Touboul, David; Thétiot-Laurent, Sophie; Pelissier, Franck; Badet-Denisot, Marie-Ange; Badet, Bernard; Durand, Philippe

    2014-08-01

    Glucosamine-6-phosphate synthase (GlmS, EC 2.6.1.16) catalyzes the first and rate-limiting step in the hexosamine biosynthetic pathway, leading to the synthesis of uridine-5'-diphospho-N-acetyl-D-glucosamine, the major building block for the edification of peptidoglycan in bacteria, chitin in fungi, and glycoproteins in mammals. This bisubstrate enzyme converts D-fructose-6-phosphate (Fru-6P) and L-glutamine (Gln) into D-glucosamine-6-phosphate (GlcN-6P) and L-glutamate (Glu), respectively. We previously demonstrated that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) allows determination of the kinetic parameters of the synthase activity. We propose here to refine the experimental protocol to quantify Glu and GlcN-6P, allowing determination of both hemisynthase and synthase parameters from a single assay kinetic experiment, while avoiding interferences encountered in other assays. It is the first time that MALDI-MS is used to survey the activity of a bisubstrate enzyme.

  9. Peptidylation for the determination of low-molecular-weight compounds by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Tang, Feng; Cen, Si-Ying; He, Huan; Liu, Yi; Yuan, Bi-Feng; Feng, Yu-Qi

    2016-05-23

    Determination of low-molecular-weight compounds by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) has been a great challenge in the analytical research field. Here we developed a universal peptide-based derivatization (peptidylation) strategy for the sensitive analysis of low-molecular-weight compounds by MALDI-TOF-MS. Upon peptidylation, the molecular weights of target analytes increase, thus avoiding serious matrix ion interference in the low-molecular-weight region in MALDI-TOF-MS. Since peptides typically exhibit good signal response during MALDI-TOF-MS analysis, peptidylation endows high detection sensitivities of low-molecular-weight analytes. As a proof-of-concept, we analyzed low-molecular-weight compounds of aldehydes and thiols by the developed peptidylation strategy. Our results showed that aldehydes and thiols can be readily determined upon peptidylation, thus realizing the sensitive and efficient determination of low-molecular-weight compounds by MALDI-TOF-MS. Moreover, target analytes also can be unambiguously detected in biological samples using the peptidylation strategy. The established peptidylation strategy is a universal strategy and can be extended to the sensitive analysis of various low-molecular-weight compounds by MALDI-TOF-MS, which may be potentially used in areas such as metabolomics.

  10. Cultivable Methylobacterium species diversity in rice seeds identified with whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis.

    Science.gov (United States)

    Okumura, Marie; Fujitani, Yoshiko; Maekawa, Masahiko; Charoenpanich, Jittima; Murage, Hunja; Kimbara, Kazuhide; Sahin, Nurettin; Tani, Akio

    2017-02-01

    Methylobacterium species are methylotrophic bacteria that widely inhabit plant surfaces. In addition to studies on methylotrophs as model organisms, research has also been conducted on their mechanism of plant growth promotion as well as the species-species specificity of plant-microbe interaction. We employed whole-cell matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (WC-MS) analysis, which enables the rapid and accurate identification of bacteria at the species level, to identify Methylobacterium isolates collected from the rice seeds of different cultivars harvested in Japan, Thailand, and Kenya. Rice seeds obtained from diverse geographical locations showed different communities of Methylobacterium species. We found that M. fujisawaense, M. aquaticum, M. platani, and M. radiotolerans are the most frequently isolated species, but none were isolated as common species from 18 seed samples due to the highly biased communities in some samples. These findings will contribute to the development of formulations containing selected species that promote rice growth, though it may be necessary to customize the formulations depending on the cultivars and farm conditions.

  11. Identification of different respiratory viruses, after a cell culture step, by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS)

    Science.gov (United States)

    Calderaro, Adriana; Arcangeletti, Maria Cristina; Rodighiero, Isabella; Buttrini, Mirko; Montecchini, Sara; Vasile Simone, Rosita; Medici, Maria Cristina; Chezzi, Carlo; De Conto, Flora

    2016-01-01

    In this study matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS), a reliable identification method for the diagnosis of bacterial and fungal infections, is presented as an innovative tool to investigate the protein profile of cell cultures infected by the most common viruses causing respiratory tract infections in humans. MALDI-TOF MS was applied to the identification of influenza A and B viruses, adenovirus C species, parainfluenza virus types 1, 2 and 3, respiratory syncytial virus, echovirus, cytomegalovirus and metapneumovirus. In this study MALDI-TOF MS was proposed as a model to be applied to the identification of cultivable respiratory viruses using cell culture as a viral proteins enrichment method to the proteome profiling of virus infected and uninfected cell cultures. The reference virus strains and 58 viruses identified from respiratory samples of subjects with respiratory diseases positive for one of the above mentioned viral agents by cell culture were used for the in vitro infection of suitable cell cultures. The isolated viral particles, concentrated by ultracentrifugation, were used for subsequent protein extraction and their spectra profiles were generated by MALDI-TOF MS analysis. The newly created library allowed us to discriminate between uninfected and respiratory virus infected cell cultures. PMID:27786297

  12. Influence of secondary structure on in-source decay of protein in matrix-assisted laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Takayama, Mitsuo; Osaka, Issey; Sakakura, Motoshi

    2012-01-01

    The susceptibility of the N-Cα bond of the peptide backbone to specific cleavage by in-source decay (ISD) in matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) was studied from the standpoint of the secondary structure of three proteins. A naphthalene derivative, 5-amino-1-naphtol (5,1-ANL), was used as the matrix. The resulting c'-ions, which originate from the cleavage at N-Cα bonds in flexible secondary structures such as turn and bend, and are free from intra-molecular hydrogen-bonded α-helix structure, gave relatively intense peaks. Furthermore, ISD spectra of the proteins showed that the N-Cα bonds of specific amino acid residues, namely Gly-Xxx, Xxx-Asp, and Xxx-Asn, were more susceptible to MALDI-ISD than other amino acid residues. This is in agreement with the observation that Gly, Asp and Asn residues usually located in turns, rather than α-helix. The results obtained indicate that protein molecules embedded into the matrix crystal in the MALDI experiments maintain their secondary structures as determined by X-ray crystallography, and that MALDI-ISD has the capability for providing information concerning the secondary structure of protein.

  13. Proteomic profiling of hepatitis B virus-related hepatocellular carcinoma with magnetic bead-based matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    Taotao Liu; Ruyi Xue; Xiaowu Huang; Danying Zhang; Ling Dong; Hao Wu; Xizhong Shen

    2011-01-01

    Proteomic techniques are promising strategies in the surveillance of hepatocellular carcinoma (HCC). This study aimed to investigate the serum profiling with magnetic bead (MB) and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and to further identify the biomarkers for HCC. Serum samples from 80 chronic hepatitis B (CHB) patients, 94 HCC concomitant with HBV patients and 24 healthy subjects were examined by MALDI-TOF MS after peptide enrichment on MBs. Based on the genetic algorithm,diagnostic models for HCC were established between 30HCC patients and 24 healthy subjects/30 CHB patients.Validations were done with the remaining cases. Markers in the models were identified through liquid chromatography (LC)/MS-MS. The three groups were well separated from each other and two discrimination models were established for HCC. The overall recognition capability of these two models was 96.25% and 93.33%, respectively.Validations showed the misdiagnosis ratio for HCC was 1.6% and 23.4%, respectively. The identified biomarkers for HCC included prothrombin precursor (fragment),calcium-dependent secretion activator 1, Baculoviral inhibitor of apoptosis repeat-containing protein 6, etc.MB-based MALDI-TOF MS is applicable in identifying the serum biomarkers and can be used in the surveillance of HCC among HBV-infected patients.

  14. Differentiation of Lactobacillus brevis strains using Matrix-Assisted-Laser-Desorption-Ionization-Time-of-Flight Mass Spectrometry with respect to their beer spoilage potential.

    Science.gov (United States)

    Kern, Carola C; Vogel, Rudi F; Behr, Jürgen

    2014-06-01

    Lactobacillus (L.) brevis is one of the most frequently encountered bacteria in beer-spoilage incidents. As the species Lactobacillus brevis comprises strains showing varying ability to grow in beer, ranging from growth in low hopped wheat to highly hopped pilsner beer, differentiation and classification of L. brevis with regard to their beer-spoiling ability is of vital interest for the brewing industry. Matrix-Assisted-Laser-Desorption-Ionization-Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) has been shown as a powerful tool for species and sub-species differentiation of bacterial isolates and is increasingly used for strain-level differentiation. Seventeen L. brevis strains, representative of different spoilage types, were characterized according to their tolerance to iso-alpha-acids and their growth in wheat-, lager- and pilsner beer. MALDI-TOF MS spectra were acquired to perform strain-level identification, cluster analysis and biomarker detection. Strain-level identification was achieved in 90% out of 204 spectra. Misidentification occurred nearly exclusively among strains belonging to the same spoilage type. Though spectra of strongly beer-spoiling strains showed remarkable similarity, no decisive single markers were detected to be present in all strains of one group. However, MALDI-TOF MS spectra can be reliably assigned to the corresponding strain and thus allow to track single strains and connect them to their physiological properties.

  15. Rapid detection of high-risk Enterococcus faecium clones by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Freitas, Ana R; Sousa, Clara; Novais, Carla; Silva, Liliana; Ramos, Helena; Coque, Teresa M; Lopes, João; Peixe, Luísa

    2017-04-01

    We aimed to explore the potential of matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for early identification of dominant Enterococcus faecium (Efm) clones involved in human infections. Well-characterized Efm isolates (n=77), analyzed by pulsed-field gel electrophoresis and multilocus sequence typing(eBURST and BAPS [Bayesian analysis of population structure] algorithms), and belonging to different hospital (n=53) and community (n=24) phylogenomic groups, were tested. Mass spectra (Bruker) were analyzed by visual inspection and different chemometric tools. Discrimination between groups comprising isolates commonly found in hospitals (BAPS 2.1a, 3.3a1, 3.3a2) and community (BAPS 2.1b and 3.2) was achieved with >99% accuracy, while identification of sequence types belonging to different BAPS subgroups was associated with >95% correct predictions. Our work is a proof of concept with regard to the suitability of MALDI-TOF MS in the identification of high-risk Efm clones. Further studies including strains from a wider variety of clones and sources will strengthen the potential of the workflow here described.

  16. Localization of ginsenosides in Panax ginseng with different age by matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry imaging.

    Science.gov (United States)

    Bai, Hangrui; Wang, Shujuan; Liu, Jianjun; Gao, Dan; Jiang, Yuyang; Liu, Hongxia; Cai, Zongwei

    2016-07-15

    The root of Panax ginseng C.A. Mey. (P. ginseng) is one of the most popular traditional Chinese medicines, with ginsenosides as its main bioactive components. Because different ginsenosides have varied pharmacological effects, extraction and separation of ginsenosides are usually required for the investigation of pharmacological effects of different ginsenosides. However, the contents of ginsenosides vary with the ages and tissues of P. ginseng root. In this research, an efficient method to explore the distribution of ginsenosides and differentiate P. ginseng roots with different ages was developed based on matrix assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-TOF-MSI). After a simple sample preparation, there were 18 peaks corresponding to 31 ginsenosides with distinct localization in the mass range of m/z 700-1400 identified by MALDI-TOF-MSI and MALDI-TOF-MS/MS. All the three types of ginsenosides were successfully detected and visualized in images, which could be correlated with anatomical features. The P. ginseng at the ages of 2, 4 and 6 could be differentiated finely through the principal component analysis of data collected from the cork based on the ion images but not data from the whole tissue. The experimental result implies that the established method for the direct analysis of metabolites in plant tissues has high potential for the rapid identification of metabolites and analysis of their localizations in medicinal herbs. Furthermore, this technique also provides valuable information for the component-specific extraction and pharmacological research of herbs.

  17. Dithranol as a matrix for matrix assisted laser desorption/ionization imaging on a fourier transform ion cyclotron resonance mass spectrometer.

    Science.gov (United States)

    Le, Cuong H; Han, Jun; Borchers, Christoph H

    2013-11-26

    Mass spectrometry imaging (MSI) determines the spatial localization and distribution patterns of compounds on the surface of a tissue section, mainly using MALDI (matrix assisted laser desorption/ionization)-based analytical techniques. New matrices for small-molecule MSI, which can improve the analysis of low-molecular weight (MW) compounds, are needed. These matrices should provide increased analyte signals while decreasing MALDI background signals. In addition, the use of ultrahigh-resolution instruments, such as Fourier transform ion cyclotron resonance (FTICR) mass spectrometers, has the ability to resolve analyte signals from matrix signals, and this can partially overcome many problems associated with the background originating from the MALDI matrix. The reduction in the intensities of the metastable matrix clusters by FTICR MS can also help to overcome some of the interferences associated with matrix peaks on other instruments. High-resolution instruments such as the FTICR mass spectrometers are advantageous as they can produce distribution patterns of many compounds simultaneously while still providing confidence in chemical identifications. Dithranol (DT; 1,8-dihydroxy-9,10-dihydroanthracen-9-one) has previously been reported as a MALDI matrix for tissue imaging. In this work, a protocol for the use of DT for MALDI imaging of endogenous lipids from the surfaces of mammalian tissue sections, by positive-ion MALDI-MS, on an ultrahigh-resolution hybrid quadrupole FTICR instrument has been provided.

  18. Direct Analysis of hCGβcf Glycosylation in Normal and Aberrant Pregnancy by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Ray K. Iles

    2014-06-01

    Full Text Available The analysis of human chorionic gonadotropin (hCG in clinical chemistry laboratories by specific immunoassay is well established. However, changes in glycosylation are not as easily assayed and yet alterations in hCG glycosylation is associated with abnormal pregnancy. hCGβ-core fragment (hCGβcf was isolated from the urine of women, pregnant with normal, molar and hyperemesis gravidarum pregnancies. Each sample was subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS analysis following dithiothreitol (DTT reduction and fingerprint spectra of peptide hCGβ 6–40 were analyzed. Samples were variably glycosylated, where most structures were small, core and largely mono-antennary. Larger single bi-antennary and mixtures of larger mono-antennary and bi-antennary moieties were also observed in some samples. Larger glycoforms were more abundant in the abnormal pregnancies and tri-antennary carbohydrate moieties were only observed in the samples from molar and hyperemesis gravidarum pregnancies. Given that such spectral profiling differences may be characteristic, development of small sample preparation for mass spectral analysis of hCG may lead to a simpler and faster approach to glycostructural analysis and potentially a novel clinical diagnostic test.

  19. Characterization of Staphylococcus aureus Isolated from Clinical Specimens by Matrix Assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    WANG Ye Ru; CHEN Qian; CUI Sheng Hui; LI Feng Qin

    2013-01-01

    Objective To develop a matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) approach to identify Staphylococcus aureus (S. aureus) and differentiate methicillin-resistant S. aureus (MRSA) from methicillin-sensitive S. aureus (MSSA). Methods A total of 100 S. aureus strains isolated from clinical specimens and farm workers were collected and analyzed by MALDI-TOF-MS. And data obtained were interpreted with biotyper software. Results Ninety-two strains were identified by MALDI-TOF-MS as S. aureus at a level of secure genus and probable species, and 4 strains were identified at probable genus after their cultivation, spectral collection and data preprocessing. One strain was identified as S. aureus with lower score. It was revealed that identification of S. aureus by MALDI-TOF-MS was highly correlated with typing by biochemical and serological methods with an accuracy as high as 97%. The biotyper cluster analysis showed that 100 isolates were divided into 2 types at the distance level of 400. Higher peak intensity in the mass of both 3784 Da and 5700 Da was observed in MRSA, whereas that was absent from MSSA. Conclusion MALDI-TOF-MS is considered as a simple, rapid and highly reproducible technique with high-throughput and accuracy for the identification of S. aureus and it can reliably differentiate MRSA from MSSA.

  20. Matrix-assisted laser desorption ionization-time of flight mass spectrometry based identification of Edwardsiella ictaluri isolated from Vietnamese striped catfish (Pangasius hypothalamus)

    Science.gov (United States)

    Nhu, Truong Quynh; Park, Seong Bin; Kim, Si Won; Lee, Jung Seok; Im, Se Pyeong; Lazarte, Jassy Mary S.; Seo, Jong Pyo; Lee, Woo-Jai; Kim, Jae Sung

    2016-01-01

    Edwardsiella (E.) ictaluri is a major bacterial pathogen that affects commercially farmed striped catfish (Pangasius hypothalamus) in Vietnam. In a previous study, 19 strains of E. ictaluri collected from striped catfish were biochemically identified with an API-20E system. Here, the same 19 strains were used to assess the ability of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS; applied using a MALDI Biotyper) to conduct rapid, easy and accurate identification of E. ictaluri. MALDI-TOF MS could directly detect the specific peptide patterns of cultured E. ictaluri colonies with high (> 2.0, indicating species-level identification) scores. MALDI Biotyper 3.0 software revealed that all of the strains examined in this study possessed highly similar peptide peak patterns. In addition, electrophoresis (SDS-PAGE) and subsequent immuno-blotting using a specific chicken antibody (IgY) against E. ictaluri revealed that the isolates had highly similar protein profiles and antigenic banding profiles. The results of this study suggest that E. ictaluri isolated from striped catfish in Vietnam have homologous protein compositions. This is important, because it indicates that MALDI-TOF MS analysis could potentially outperform the conventional methods of identifying E. ictaluri. PMID:26726022

  1. Novel approach for differentiating Shigella species and Escherichia coli by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Khot, Prasanna D; Fisher, Mark A

    2013-11-01

    Shigella species are so closely related to Escherichia coli that routine matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) cannot reliably differentiate them. Biochemical and serological methods are typically used to distinguish these species; however, "inactive" isolates of E. coli are biochemically very similar to Shigella species and thus pose a greater diagnostic challenge. We used ClinProTools (Bruker Daltonics) software to discover MALDI-TOF MS biomarker peaks and to generate classification models based on the genetic algorithm to differentiate between Shigella species and E. coli. Sixty-six Shigella spp. and 72 E. coli isolates were used to generate and test classification models, and the optimal models contained 15 biomarker peaks for genus-level classification and 12 peaks for species-level classification. We were able to identify 90% of E. coli and Shigella clinical isolates correctly to the species level. Only 3% of tested isolates were misidentified. This novel MALDI-TOF MS approach allows laboratories to streamline the identification of E. coli and Shigella species.

  2. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry for the rapid identification of yeasts causing bloodstream infections.

    Science.gov (United States)

    Ghosh, A K; Paul, S; Sood, P; Rudramurthy, S M; Rajbanshi, A; Jillwin, T J; Chakrabarti, A

    2015-04-01

    Few studies have systematically standardised and evaluated matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for identification of yeasts from bloodstream infections. This is rapidly becoming pertinent for early identification of yeasts and appropriate antifungal therapy. We used 354 yeast strains identified by polymerase chain reaction (PCR) sequencing for standardisation and 367 blind clinical strains for validation of our MALDI-TOF MS protocols. We also evaluated different sample preparation methods and found the on-plate formic acid extraction method as most cost- and time-efficient. The MALDI-TOF assay correctly identified 98.9% of PCR-sequenced yeasts. Novel main spectrum projections (MSP) were developed for Candida auris, C. viswanathii and Kodamaea ohmeri, which were missing from the Bruker MALDI-TOF MS database. Spectral cut-offs computed by receiver operating characteristics (ROC) analysis showed 99.4% to 100% accuracy at a log score of ≥ 1.70 for C. tropicalis, C. parapsilosis, C. pelliculosa, C. orthopsilosis, C. albicans, C. rugosa, C. guilliermondii, C. lipolytica, C. metapsilosis, C. nivariensis. The differences in the species-specific scores of our standardisation and blind validation strains were not statistically significant, implying the optimal performance of our test protocol. The MSPs of the three new species also were validated. We conclude that MALDI-TOF MS is a rapid, accurate and reliable tool for identification of bloodstream yeasts. With proper standardisation, validation and regular database expansion, its efficiency can be further enhanced.

  3. Application of porous metal enrichment probe sampling to single cell analysis using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).

    Science.gov (United States)

    Fu, Qiang; Tang, Jun; Cui, Meng; Xing, Junpeng; Liu, Zhiqiang; Liu, Shuying

    2016-01-01

    There is an increasing need for analyzing metabolism in a single cell, which is important to understand the nature of cellular heterogeneity, disease, growth and specialization, etc. However, single cell analysis is often challenging for the traces of samples. In the present study, porous metal enrichment probe sampling combined with matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) has been applied for in situ analysis of live onion epidemic cell. Porous probe, treated by corroding copper wire with HCl, was directly inserted into a single cell to get cell solution. A self-made linear actuator was enough to control the penetration of probe into the target cell accurately. Then samples on the tip of probe were eluted and detected by a commercial MALDI-TOF-MS directly. The formation of porous microstructure on the probe surface increased the adsorptive capacity of cell solution. The sensitivity of porous probe sampling was 6 times higher than uncorroded probes generally. This method provides a sensitive and convenient way for the sampling and detection of single cell solution. Copyright © 2015 John Wiley & Sons, Ltd.

  4. A novel cluster of Mycobacterium abscessus complex revealed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS).

    Science.gov (United States)

    Suzuki, Hiromichi; Yoshida, Shiomi; Yoshida, Atsushi; Okuzumi, Katsuko; Fukusima, Atsuhito; Hishinuma, Akira

    2015-12-01

    Mycobacterium abscessus complex is a rapidly growing mycobacterium consisting of 3 subspecies, M. abscessus, Mycobacterium massiliense, and Mycobacterium bolletii. However, rapid and accurate species identification is difficult. We first evaluated a suitable protocol of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for distinguishing these subspecies. Then, we studied spectral signals by MALDI-TOF MS in 59 M. abscessus, 42 M. massiliense, and 2 M. bolletii. Among several specific spectral signals, 4 signals clearly differentiate M. massiliense from the other 2 subspecies, M. abscessus and M. bolletii. Moreover, 6 of the 42 M. massiliense isolates showed a spectral pattern similar to M. abscessus. These isolates correspond to the distinctive class of M. massiliense (cluster D) which is closer to M. abscessus by the previous variable number tandem repeat analysis. These results indicate that MALDI-TOF MS is not only useful for the identification of 3 subspecies of M. abscessus complex but also capable of distinguishing clusters of M. massiliense.

  5. Enhanced reliability of avian influenza virus (AIV) and Newcastle disease virus (NDV) identification using matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS).

    Science.gov (United States)

    Jang, Ho Bin; Sung, Haan Woo; Nho, Seong Won; Park, Seong Bin; Cha, In Seok; Aoki, Takashi; Jung, Tae Sung

    2011-03-01

    In-solution enzymatic and nonenzymatic digestion methods have been successfully implemented in matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS)-based virus identification, extending to typing/subtyping of deadly influenza viruses. However, these methods are inefficient in obtaining more precise information on surface proteins of myxovirus particles, not only the hemagglutinin and neuraminidase of influenza virus but also the hemagglutinin-neuraminidase of Newcastle disease virus (NDV). Imbalances in viral protein composition cause ion suppression of tryptic fragments from low-abundant target proteins (surface proteins), adversely affecting reproducibility of mass spectra. Additionally, the coexistence of tryptic peptides from several proteins requires sophisticated statistical solutions for precise result interpretations. To circumvent these, we apply detergent-based (gel-free) partitioning of whole viruses into soluble surface proteins and insoluble virus materials, using differential centrifugation. MALDI-TOF or MALDI-TOF/TOF MS was applied to analyze tryptic peptides from separated viral proteins. In this study, we achieved type/subtype of avian influenza virus (AIV) within 5 h, based on 4 major proteins, by significantly reducing ion suppression and signal overlap from various protein sources. Hence, our approach can both yield dependable results and allow Web-based search engines to be directly employed, obviating the need for additional statistical strategy. Additionally, we demonstrate the utility of the method using NDV.

  6. Independent assessment of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) sample preparation quality: A novel statistical approach for quality scoring.

    Science.gov (United States)

    Kooijman, Pieter C; Kok, Sander J; Weusten, Jos J A M; Honing, Maarten

    2016-05-05

    Preparation of samples according to an optimized method is crucial for accurate determination of polymer sample characteristics by Matrix-Assisted Laser Desorption Ionization (MALDI) analysis. Sample preparation conditions such as matrix choice, cationization agent, deposition technique or even the deposition volume should be chosen to suit the sample of interest. Many sample preparation protocols have been developed and employed, yet finding the optimal sample preparation protocol remains a challenge. Because an objective comparison between the results of diverse protocols is not possible, "gut-feeling" or "good enough" is often decisive in the search for an optimum. This implies that sub-optimal protocols are used, leading to a loss of mass spectral information quality. To address this problem a novel analytical strategy based on MALDI imaging and statistical data processing was developed in which eight parameters were formulated to objectively quantify the quality of sample deposition and optimal MALDI matrix composition and finally sum up to an overall quality score of the sample deposition. These parameters can be established in a fully automated way using commercially available mass spectrometry imaging instruments without any hardware adjustments. With the newly developed analytical strategy the highest quality MALDI spots were selected, resulting in more reproducible and more valuable spectra for PEG in a variety of matrices. Moreover, our method enables an objective comparison of sample preparation protocols for any analyte and opens up new fields of investigation by presenting MALDI performance data in a clear and concise way.

  7. An evaluation of matrix-assisted laser desorption ionization time-of-flight mass spectrometry for the identification of Staphylococcus pseudintermedius isolates from canine infections.

    Science.gov (United States)

    Silva, Marcella Braga; Ferreira, Fabienne Antunes; Garcia, Luize Neli Nunes; Silva-Carvalho, Maria Cícera; Botelho, Larissa Alvarenga Batista; Figueiredo, Agnes Marie Sá; Vieira-da-Motta, Olney

    2015-03-01

    It has been proposed, based on taxonomic and molecular studies, that all canine isolates belonging to Staphylococcus intermedius group (SIG) should be renamed Staphylococcus pseudintermedius. However, isolates of SIG and other coagulase-positive staphylococci share many phenotypic characteristics, which could lead to misidentification. The accuracy of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for identifying S. pseudintermedius isolates obtained from canine infections was evaluated, using a polymerase chain reaction (PCR)-based identification as the gold standard. In addition, MALDI-TOF MS was compared with conventional biochemical tests. A central problem was the incorrect identification of S. pseudintermedius isolates as S. intermedius by either MALDI-TOF MS or biochemical identification. From the 49 S. pseudintermedius isolates identified by the molecular method, only 21 could be assigned to this species by the biochemical approach and only 12 by MALDI-TOF MS. The 6 S. aureus isolates were correctly identified by all 3 techniques. However, using biochemical tests, 9 S. pseudintermedius were mistakenly classified as S. aureus, indicating a reduced specificity relative to the MALDI-TOF MS system. Analysis with the MALDI-TOF MS platform allowed rapid and accurate identification of the 49 isolates to the S. intermedius group but the approach was very limited in identifying S. pseudintermedius isolates, as only 12 of 49 isolates were correctly identified, a sensitivity of 0.24 (95% confidence interval: 0.13-0.39).

  8. Paraffin-wax-coated plates as matrix-assisted laser desorption/ionization sample support for high-throughput identification of proteins by peptide mass fingerprinting.

    Science.gov (United States)

    Tannu, Nilesh S; Wu, Jian; Rao, Vamshi K; Gadgil, Himanshu S; Pabst, Michael J; Gerling, Ivan C; Raghow, Rajendra

    2004-04-15

    We compared trysin-digested protein samples desalted by ZipTip(C18) reverse-phase microcolumns with on-plate washing of peptides deposited either on paraffin-coated plates (PCP), Teflon-based AnchorChip plates, or stainless steel plates, before analysis by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). Trypsinized bovine serum albumin and ovalbumin and 16 protein spots extracted from silver-stained two-dimensional gels of murine C(2)C(12) myoblasts or human leukocytes, prepared by the above two methods, were subjected to MALDI on PCP, AnchorChip plates, or uncoated stainless steel plates. Although most peptide mass peaks were identical regardless of the method of desalting and concentrating of protein samples, samples washed and concentrated by the PCP-based method had peptide peaks that were not seen in the samples prepared using the ZipTip(C18) columns. The mass spectra of peptides desalted and washed on uncoated stainless steel MALDI plates were consistently inferior due to loss of peptides. Some peptides of large molecular masses were apparently lost from samples desalted by ZipTip(C18) microcolumns, thus diminishing the quality of the fingerprint needed for protein identification. We demonstrate that the method of washing of protein samples on paraffin-coated plates provides an easy, reproducible, inexpensive, and high-throughput alternative to ZipTip(C18)-based purification of protein prior to MALDI-TOF-MS analysis.

  9. Hydrophilic interaction chromatography coupled matrix assisted laser desorption/ionization mass spectrometry for molecular analysis of organic compounds in medicines, tea, and coffee

    KAUST Repository

    Wang, Renqi

    2013-01-01

    Natural occurring organic compounds from food, natural organic matter, as well as metabolic products have received intense attention in current chemical and biological studies. Examination of unknown compounds in complex sample matrices is hampered by the limited choices for data readout and molecular elucidation. Herein, we report a generic method of hydrophilic interaction chromatography (HILIC) coupled with matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) for the rapid characterization of ingredients in pharmaceutical compounds, tea, and coffee. The analytes were first fractionated using a cationic HILIC column prior to MALDI-MS analyses. It was found that the retention times of a compound arising from different samples were consistent under the same conditions. Accordingly, molecules can be readily characterized by both the mass and chromatographic retention time. The retention behaviors of acidic and basic compounds on the cationic HILIC column were found to be significantly influenced by the pH of mobile phases, whereas neutral compounds depicted a constant retention time at different pH. The general HILIC-MALDI-MS method is feasible for fast screening of naturally occurring organic compounds. A series of homologs can be determined if they have the same retention behavior. Their structural features can be elucidated by considering their mass differences and hydrophilic properties as determined by HILIC chromatogram. © 2013 The Royal Society of Chemistry.

  10. Enhanced visualization of small peptides absorbed in rat small intestine by phytic-acid-aided matrix-assisted laser desorption/ionization-imaging mass spectrometry.

    Science.gov (United States)

    Hong, Seong-Min; Tanaka, Mitsuru; Yoshii, Saori; Mine, Yoshinori; Matsui, Toshiro

    2013-11-05

    Enhanced visualization of small peptides absorbed through a rat intestinal membrane was achieved by matrix-assisted laser desorption/ionization time-of-flight imaging mass spectrometry (MALDI-IMS) with the aid of phytic acid as a matrix additive. Penetrants through intestinal peptide transporter 1, i.e., glycyl-sarcosine (Gly-Sar, 147.1 m/z) and antihypertensive dipeptide, Val-Tyr (281.2 m/z), were chosen for MALDI-IMS. The signal-to-noise (S/N) ratios of dipeptides Gly-Sar and Val-Tyr were seen to increase by 2.4- and 8.0-fold, respectively, when using a 2',4',6'-trihydroxyacetophenone (THAP) matrix containing 5.0 mM phytic acid, instead of the THAP matrix alone. Owing to the phytic-acid-aided MALDI-IMS method, Gly-Sar and Val-Tyr absorbed in the rat intestinal membrane were successfully visualized. The proposed imaging method also provided useful information on intestinal peptide absorption; to some extent, Val-Tyr was rapidly hydrolyzed to Tyr by peptidases located at the intestinal microvillus during the absorption process. In conclusion, the strongly acidic additive, phytic acid, is beneficial for enhancing the visualization of small peptides using MALDI-IMS, owing to the suppression of ionization-interfering salts in the tissue.

  11. Evaluation of a Semiquantitative Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Method for Rapid Antimicrobial Susceptibility Testing of Positive Blood Cultures.

    Science.gov (United States)

    Jung, Jette S; Hamacher, Christina; Gross, Birgit; Sparbier, Katrin; Lange, Christoph; Kostrzewa, Markus; Schubert, Sören

    2016-11-01

    With the increasing prevalence of multidrug-resistant Gram-negative bacteria, rapid identification of the pathogen and its individual antibiotic resistance is crucial to ensure adequate antiinfective treatment at the earliest time point. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry for the identification of bacteria directly from the blood culture bottle has been widely established; however, there is still an urgent need for new methods that permit rapid resistance testing. Recently, a semiquantitative MALDI-TOF mass spectrometry-based method for the prediction of antibiotic resistance was described. We evaluated this method for detecting nonsusceptibility against two β-lactam and two non-β-lactam antibiotics. A collection of 30 spiked blood cultures was tested for nonsusceptibility against gentamicin and ciprofloxacin. Furthermore, 99 patient-derived blood cultures were tested for nonsusceptibility against cefotaxime, piperacillin-tazobactam, and ciprofloxacin in parallel with MALDI-TOF mass spectrometry identification from the blood culture fluid. The assay correctly classified all isolates tested for nonsusceptibility against gentamicin and cefotaxime. One misclassification for ciprofloxacin nonsusceptibility and five misclassifications for piperacillin-tazobactam nonsusceptibility occurred. Identification of the bacterium and prediction of nonsusceptibility was possible within approximately 4 h.

  12. Identification of pathogenic microorganisms directly from positive blood vials by matrix-assisted laser desorption/ionization time of flight mass spectrometry

    DEFF Research Database (Denmark)

    Nonnemann, Bettina; Tvede, Michael; Bjarnsholt, Thomas

    2013-01-01

    Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) is a promising and fast method for identifying fungi and bacteria directly from positive blood cultures. Various pre-treatment methods for MALDI-TOF MS identification have been reported for this purpose. In......-house results for identification of bacterial colonies by MALDI-TOF MS using a cut-off score of 1.5 did not reduce the diagnostic accuracy compared with the recommended cut-off score of 1.8. A 3-month consecutive study of positive blood cultures was carried out in our laboratory to evaluate whether...... the Sepsityper™ Kit (Bruker Daltonics) with Biotyper 2.0 software could be used as a fast diagnostic tool for bacteria and fungi and whether a 1.5 cut-off score could improve species identification compared with the recommended score of 1.8. Two hundred and fifty-six positive blood vials from 210 patients and 19...

  13. Turnaround time of positive blood cultures after the introduction of matrix-assisted laser desorption-ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Angeletti, Silvia; Dicuonzo, Giordano; D'Agostino, Alfio; Avola, Alessandra; Crea, Francesca; Palazzo, Carlo; Dedej, Etleva; De Florio, Lucia

    2015-07-01

    A comparative evaluation of the turnaround time (TAT) of positive blood culture before and after matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) introduction in the laboratory routine was performed. A total of 643 positive blood cultures, of which 310 before and 333 after MALDI-TOF technique introduction, were collected. In the post MALDI-TOF period, blood culture median TAT decreased from 73.53 hours to 71.73 for Gram-positive, from 64.09 hours to 63.59 for Gram-negative and from 115.7 hours to 47.62 for anaerobes. MALDI-TOF significantly decreased the TAT of anaerobes, for which antimicrobial susceptibility test is not routinely performed. Furthermore, the major advantage of MALDI-TOF introduction was the decrease of the time for pathogen identification (TID) independently from the species with an improvement of 93% for Gram-positive, 86% for Gram-negative and 95% for anaerobes. In addition, high species-level identification rates and cost savings than conventional methods were achieved after MALDI-TOF introduction.

  14. Gold-decorated titania nanotube arrays as dual-functional platform for surface-enhanced Raman spectroscopy and surface-assisted laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Nitta, Syuhei; Yamamoto, Atsushi; Kurita, Masahiro; Arakawa, Ryuichi; Kawasaki, Hideya

    2014-06-11

    In this report, we demonstrate gold-decorated titania nanotube arrays (Au-TNA substrate) as a dual-functional platform for surface-enhanced Raman spectroscopy (SERS) and surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). The Au nanoparticles are grown on the substrate using vapor deposition of Au. The resulting substrates perform better than Au colloids in terms of the reproducibility of the SERS measurements, long-term stability of the fabricated structures, and clean surface of the Au. The nanostructure of the Au-TNA substrate was designed to optimize the SALDI-MS and SERS performance. Excellent reproducibility of the SERS measurements using the Au-TNA substrate was obtained, with a standard error less than 6 %. SALDI activity was also demonstrated for the same Au-TNA substrates. Finally, the Au-TNA substrate was used for combined SERS and SALDI-MS analysis (i) to discriminate the structural isomers of pyridine compounds (para-, meta-, and ortho-pyridinecarboxylic acid) and (ii) to detect polycarbamate, a dithiocarbamate fungicide. These results are difficult to obtain using either approach alone.

  15. Accuracy of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of clinical pathogenic fungi: a meta-analysis.

    Science.gov (United States)

    Ling, Huazhi; Yuan, Zhijie; Shen, Jilu; Wang, Zhongxin; Xu, Yuanhong

    2014-07-01

    Fungal infections in the clinic have become increasingly serious. In many cases, the identification of clinically relevant fungi remains time-consuming and may also be unreliable. Matrix-assisted laser desorption ionization-time of flight mass spectroscopy (MALDI-TOF MS) is a newly developed diagnostic tool that is increasingly being employed to rapidly and accurately identify clinical pathogenic microorganisms. The present meta-analysis aimed to systematically evaluate the accuracy of MALDI-TOF MS for the identification of clinical pathogenic fungi. After a rigorous selection process, 33 articles, involving 38 trials and a total of 9,977 fungal isolates, were included in the meta-analysis. The random-effects pooled identification accuracy of MALDI-TOF MS increased from 0.955 (95% confidence interval [CI], 0.939 to 0.969) at the species level to 0.977 (95% CI, 0.955 to 0.993) at the genus level (P meta-analysis and some of the subanalyses. In parallel, significant differences in heterogeneity among different systems and among different methods for calculating the identification ratios were found by multivariate metaregression, but none of the factors, except for the moderator of outcome, was significantly associated with heterogeneity by univariate metaregression. In summary, the MALDI-TOF MS method is highly accurate for the identification of clinically pathogenic fungi; future studies should analyze the comprehensive capability of this technology for clinical diagnostic microbiology.

  16. Monitoring the enzymatic polymerization of 4-phenylphenol by matrix-assisted laser desorption ionization time-of-flight mass spectrometry: a novel approach.

    Science.gov (United States)

    Xu, Peng; Kumar, Jayant; Samuelson, Lynne; Cholli, Ashok L

    2002-01-01

    Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry is a powerful tool for polymer characterization. It has been used to understand the enzymatic polymerization of 4-phenylphenol and to monitor number average molecular weight and weight average molecular weight of the polymer as a function of systematic addition of hydrogen peroxide (H(2)O(2)) in the reaction. A novel method, an introduction of internal standard for quantification of data, has been developed for MALDI-TOF MS to investigate the fate of each mers during the reaction. The preliminary data suggest that this approach provides new insight on the enzymatic synthesis, which is not available by other techniques. For the first time, we are able to understand the fate of several mers as a function of reaction conditions. The relative content of each mer increases with the addition of H(2)O(2), except for dimer and trimer. For example, the concentration of dimer species decreases as a function of H(2)O(2). On the other hand, the concentration of trimer species increases first and then decreases in the course of the reaction.

  17. Detection of bacteria from biological mixtures using immunomagnetic separation combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Science.gov (United States)

    Madonna, A.J.; Basile, F.; Furlong, E.; Voorhees, K.J.

    2001-01-01

    A rapid method for identifying specific bacteria from complex biological mixtures using immunomagnetic separation coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been developed. The technique employs commercially available magnetic beads coated with polycolonal antibodies raised against specific bacteria and whole cell analysis by MALDI-MS. A suspension of a bacterial mixture is mixed with the immunomagnetic beads specific for the target microorganism. After a short incubation period (20 mins) the bacteria captured by the beads are washed, resuspended in deionized H2O and directly applied onto a MALDI probe. Liquid suspensions containing bacterial mixtures can be screened within 1 h total analysis time. Positive tests result in the production of a fingerprint mass spectrum primarily consisting of protein biomarkers characteristic of the targeted microorganism. Using this procedure, Salmonella choleraesuis was isolated and detected from standard bacterial mixtures and spiked samples of river water, human urine, and chicken blood. Copyright ?? 2001 John Wiley & Sons, Ltd.

  18. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry profiling of trace constituents of condom lubricants in the presence of biological fluids.

    Science.gov (United States)

    Spencer, Sandra E; Kim, Sin Young; Kim, Seoung Bum; Schug, Kevin A

    2011-04-15

    The use of condoms in sexual assault cases has become increasingly common due to the heightened awareness of the use of DNA as evidence in criminal investigations. The ability to identify and differentiate the polymers and additives found in lubricant residues can provide investigators leads and insights as to the perpetrator of a sexual assault. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) is ideal for detecting condom lubricants and additives; the instrument is capable of surveying analytes across a wide mass range and is a preferred technique for the analysis of polymers. Three MALDI-TOF-MS methods directed toward the detection and differentiation of condom and personal lubricant residues, as well as their mixtures with biological fluids, were developed and compared: (a) a sample premixed with aqueous matrix; (b) a sample premixed with an ionic liquid matrix; and (c) a layering method that incorporates a cationization reagent. Of the three, the layered method that utilized sodium chloride as a cationization reagent showed the best sensitivity and selectivity. This method allowed for the segregation of the various lubricant formulas into a discrete number of groups. Infrared spectroscopy was used to support and clarify the MALDI data. Principal component analysis was used to further demonstrate the ability of this method to segregate various lubricant types into a limited number of classes. Additionally, lubricant residues could be detected in the presence of biological fluids down to a fraction of a percent.

  19. Flexible xxx-asp/asn and gly-xxx residues of equine cytochrome C in matrix-assisted laser desorption/ionization in-source decay mass spectrometry.

    Science.gov (United States)

    Takayama, Mitsuo

    2012-01-01

    The backbone flexibility of a protein has been studied from the standpoint of the susceptibility of amino acid residues to in-source decay (ISD) in matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). Residues more susceptible to MALDI-ISD, namely Xxx-Asp/Asn and Gly-Xxx, were identified from the discontinuous intense peak of c'-ions originating from specific cleavage at N-Cα bonds of the backbone of equine cytochrome c. The identity of the residues susceptible to ISD was consistent with the known flexible backbone amides as estimated by hydrogen/deuterium exchange (HDX) experiments. The identity of these flexible amino acid residues (Asp, Asn, and Gly) is consistent with the fact that these residues are preferred in flexible secondary structure free from intramolecular hydrogen-bonded structures such as α-helix and β-sheet. The MALDI-ISD spectrum of equine cytochrome c gave not only intense N-terminal side c'-ions originating from N-Cα bond cleavage at Xxx-Asp/Asn and Gly-Xxx residues, but also C-terminal side complement z'-ions originating from the same cleavage sites. The present study implies that MALDI-ISD can give information about backbone flexibility of proteins, comparable with the protection factors estimated by HDX.

  20. Applications of whole-cell matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry in systematic microbiology.

    Science.gov (United States)

    Welker, Martin; Moore, Edward R B

    2011-02-01

    In the last few years matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been increasingly studied and applied for the identification and typing of microorganisms. Very recently, MALDI-TOF MS has been introduced in clinical routine microbiological diagnostics with marked success, which is remarkable considering that not long ago the technology was generally seen as being far from practical application. The identification of microbial isolates by whole-cell mass spectrometry (WC-MS) is being recognized as one of the latest tools forging a revolution in microbial diagnostics, with the potential of bringing to an end many of the time-consuming and man-power-intensive identification procedures that have been used for decades. Apart from applications of WC-MS in clinical diagnostics, other fields of microbiology also have adopted the technology with success. In this article, an over-view of the principles of MALDI-TOF MS and WC-MS is presented, highlighting the characteristics of the technology that allow its utilization for systematic microbiology.

  1. Evaluation of a simple protein extraction method for species identification of clinically relevant staphylococci by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Matsuda, Naoto; Matsuda, Mari; Notake, Shigeyuki; Yokokawa, Hirohide; Kawamura, Yoshiaki; Hiramatsu, Keiichi; Kikuchi, Ken

    2012-12-01

    In clinical microbiology, bacterial identification is labor-intensive and time-consuming. A solution for this problem is the use of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). In this study, we evaluated a modified protein extraction method of identification performed on target plates (on-plate extraction method) with MALDI-TOF (Bruker Microflex LT with Biotyper version 3.0) and compared it to 2 previously described methods: the direct colony method and a standard protein extraction method (standard extraction method). We evaluated the species of 273 clinical strains and 14 reference strains of staphylococci. All isolates were characterized using the superoxide dismutase A sequence as a reference. For the species identification, the on-plate, standard extraction, and direct colony methods identified 257 isolates (89.5%), 232 isolates (80.8%), and 173 isolates (60.2%), respectively, with statistically significant differences among the three methods (P extraction method is at least as good as standard extraction in identification rate and has the advantage of a shorter processing time.

  2. Differentiation of clinically relevant Mucorales Rhizopus microsporus and R. arrhizus by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).

    Science.gov (United States)

    Dolatabadi, Somayeh; Kolecka, Anna; Versteeg, Matthijs; de Hoog, Sybren G; Boekhout, Teun

    2015-07-01

    This study addresses the usefulness of matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS for reliable identification of the two most frequently occurring clinical species of Rhizopus, namely Rhizopus arrhizus with its two varieties, arrhizus and delemar, and Rhizopus microsporus. The test-set comprised 38 isolates of clinical and environmental origin previously identified by internal transcribed spacer (ITS) sequencing of rDNA. Multi-locus sequence data targeting three gene markers (ITS, ACT, TEF ) showed two monophylic clades for Rhizopus arrhizus and Rhizopus microsporus (bootstrap values of 99 %). Cluster analysis confirmed the presence of two distinct clades within Rhizopus arrhizus representing its varieties arrhizus and delemar. The MALDI Biotyper 3.0 Microflex LT platform (Bruker Daltonics) was used to confirm the distinction between Rhizopus arrhizus and Rhizopus microsporus and the presence of two varieties within the species Rhizopus arrhizus. An in-house database of 30 reference main spectra (MSPs) was initially tested for correctness using commercially available databases of Bruker Daltonics. By challenging the database with the same strains of which an in-house database was created, automatic identification runs confirmed that MALDI-TOF MS is able to recognize the strains at the variety level. Based on principal component analysis, two MSP dendrograms were created and showed concordance with the multi-locus tree; thus, MALDI-TOF MS is a useful tool for diagnostics of mucoralean species.

  3. Investigation of colloidal graphite as a matrix for matrix-assisted laser desorption/ionisation mass spectrometry of low molecular weight analytes.

    Science.gov (United States)

    Warren, Alexander D; Conway, Ulric; Arthur, Christopher J; Gates, Paul J

    2016-07-01

    The analysis of low molecular weight compounds by matrix-assisted laser desorption/ionisation mass spectrometry is problematic due to the interference and suppression of analyte ionisation by the matrices typically employed - which are themselves low molecular weight compounds. The application of colloidal graphite is demonstrated here as an easy to use matrix that can promote the ionisation of a wide range of analytes including low molecular weight organic compounds, complex natural products and inorganic complexes. Analyte ionisation with colloidal graphite is compared with traditional organic matrices along with various other sources of graphite (e.g. graphite rods and charcoal pencils). Factors such as ease of application, spectra reproducibility, spot longevity, spot-to-spot reproducibility and spot homogeneity (through single spot imaging) are explored. For some analytes, considerable matrix suppression effects are observed resulting in spectra completely devoid of matrix ions. We also report the observation of radical molecular ions [M(-●) ] in the negative ion mode, particularly with some aromatic analytes. Copyright © 2016 John Wiley & Sons, Ltd.

  4. Matrix-assisted laser desorption/ionization mass spectrometry imaging: a powerful tool for probing the molecular topology of plant cutin polymer.

    Science.gov (United States)

    Veličković, Dušan; Herdier, Hélène; Philippe, Glenn; Marion, Didier; Rogniaux, Hélène; Bakan, Bénédicte

    2014-12-01

    The cutin polymers of different fruit cuticles (tomato, apple, nectarine) were examined using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) after in situ release of the lipid monomers by alkaline hydrolysis. The mass spectra were acquired from each coordinate with a lateral spatial resolution of approximately 100 μm. Specific monomers were released at their original location in the tissue, suggesting that post-hydrolysis diffusion can be neglected. Relative quantification of the species was achieved by introducing an internal standard, and the collection of data was subjected to non-supervised and supervised statistical treatments. The molecular images obtained showed a specific distribution of ions that could unambiguously be ascribed to cutinized and suberized regions observed at the surface of fruit cuticles, thus demonstrating that the method is able to probe some structural changes that affect hydrophobic cuticle polymers. Subsequent chemical assignment of the differentiating ions was performed, and all of these ions could be matched to cutin and suberin molecular markers. Therefore, this MALDI-MSI procedure provides a powerful tool for probing the surface heterogeneity of plant lipid polymers. This method should facilitate rapid investigation of the relationships between cuticle phenotypes and the structure of cutin within a large population of mutants.

  5. Discrimination of Bacillus anthracis Spores by Direct in-situ Analysis of Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Jeong, Youngsu; Lee, Jonghee; Kim, Seongsoo [Agency for Defense Development, Daejeon (Korea, Republic of)

    2013-09-15

    The rapid and accurate identification of biological agents is a critical step in the case of bio-terror and biological warfare attacks. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been widely used for the identification of microorganisms. In this study, we describe a method for the rapid and accurate discrimination of Bacillus anthracis spores using MALDI-TOF MS. Our direct in-situ analysis of MALDI-TOF MS does not involve subsequent high-resolution mass analyses and sample preparation steps. This method allowed the detection of species-specific biomarkers from each Bacillus spores. Especially, B. anthracis spores had specific biomarker peaks at 2503, 3089, 3376, 6684, 6698, 6753, and 6840 m/z. Cluster and PCA analyses of the mass spectra of Bacillus spores revealed distinctively separated clusters and within-groups similarity. Therefore, we believe that this method is effective in the real-time identification of biological warfare agents such as B. anthracis as well as other microorganisms in the field.

  6. A SIMPLE AND RAPID MATRIX-ASSISTED LASER DESORPTION/IONIZATION TIME OF FLIGHT MASS SPECTROMETRY METHOD TO SCREEN FISH PLASMA SAMPLES FOR ESTROGEN-RESPONSIVE BIOMARKERS

    Science.gov (United States)

    In this study, we describe and evaluate the performance of a simple and rapid mass spectral method for screening fish plasma for estrogen-responsive biomarkers using matrix assisted laster desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) couopled with a short...

  7. Rapid Identification of Bacteria Directly from Positive Blood Cultures by Use of a Serum Separator Tube, Smudge Plate Preparation, and Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

    Science.gov (United States)

    Chen, Yan; Porter, Vanessa; Mubareka, Samira; Kotowich, Leona; Simor, Andrew E

    2015-10-01

    We analyzed the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) of smudge plate growth for bacterial identification from 400 blood cultures. Ninety-seven percent of Gram-negative bacilli and 85% of Gram-positive organisms were correctly identified within 4 h; only eight isolates (2.0%) were misidentified. This method provided rapid and accurate microbial identification from positive blood cultures.

  8. Amazonian Vegetable Oils and Fats: Fast Typification and Quality Control via Triacylglycerol (TAG) Profiles from Dry Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-TOF) Mass Spectrometry Fingerprinting

    OpenAIRE

    Saraiva, Sergio A.; Cabral, Elaine C; Eberlin, Marcos N; Rodrigo R. Catharino

    2009-01-01

    Amazonian oils and fats display unique triacylglycerol (TAG) profiles and, because of their economic importance as renewable raw materials and use by the cosmetic and food industries, are often subject to adulteration and forgery. Representative samples of these oils (andiroba, Brazil nut, buriti, and passion fruit) and fats (cupuacu, murumuru, and ucuba) were characterized without pre-separation or derivatization via dry (solvent-free) matrix-assisted laser desorption/ionization time-of-flig...

  9. Comparison of the accuracy of matrix-assisted laser desorption ionization-time of flight mass spectrometry with that of other commercial identification systems for identifying Staphylococcus saprophyticus in urine.

    Science.gov (United States)

    Lee, Tai-Fen; Lee, Hao; Chen, Chung-Ming; Du, Shin-Hei; Cheng, Ya-Chih; Hsu, Chen-Ching; Chung, Meng-Yu; Teng, Shih-Hua; Teng, Lee-Jene; Hsueh, Po-Ren

    2013-05-01

    Among 30 urinary isolates of Staphylococcus saprophyticus identified by sequencing methods, the rate of accurate identification was 100% for Bruker Biotyper matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), 86.7% for the Phoenix PID and Vitek 2 GP systems, 93.3% for the MicroScan GP33 system, and 46.7% for the BBL CHROMagar Orientation system.

  10. 1,8-Bis(dimethylamino)naphthalene/9-aminoacridine: A new binary matrix for lipid fingerprinting of intact bacteria by matrix assisted laser desorption ionization mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Calvano, C.D., E-mail: cosimadamiana.calvano@uniba.it [Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Via Orabona, 4, 70126 Bari (Italy); Monopoli, A.; Ditaranto, N. [Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Via Orabona, 4, 70126 Bari (Italy); Palmisano, F. [Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Via Orabona, 4, 70126 Bari (Italy); Centro Interdipartimentale di Ricerca S.M.A.R.T., Università degli Studi di Bari Aldo Moro, Via Orabona, 4, 70126 Bari (Italy)

    2013-10-10

    Graphical abstract: -- Highlights: •New binary matrix for less ionizable lipid analysis with no interfering peaks. •Combined MALDI and X-ray photoelectron spectroscopy (XPS) analyses. •Fast lipid fingerprint on Gram positive and Gram negative bacteria by MALDI MS. •Mapping of phospholipids by XPS imaging. •Very fast membrane lipid extraction procedure. -- Abstract: The effectiveness of a novel binary matrix composed of 1,8-bis(dimethylamino)naphthalene (DMAN; proton sponge) and 9-aminoacridine (9AA) for the direct lipid analysis of whole bacterial cells by matrix assisted laser desorption ionization mass spectrometry (MALDI MS) is demonstrated. Deprotonated analyte signals nearly free of matrix-related ions were observed in negative ion mode. The effect of the most important factors (laser energy, pulse voltage, DMAN/9AA ratio, analyte/matrix ratio) was investigated using a Box–Behnken response surface design followed by multi-response optimization in order to simultaneously maximize signal-to-noise (S/N) ratio and resolution. The chemical surface composition of single or mixed matrices was explored by X-ray photoelectron spectroscopy (XPS). Moreover, XPS imaging was used to map the spatial distribution of a model phospholipid in single or binary matrices. The DMAN/9AA binary matrix was then successfully applied to the analysis of intact Gram positive (Lactobacillus sanfranciscensis) or Gram negative (Escherichia coli) microorganisms. About fifty major membrane components (free fatty acids, mono-, di- and tri-glycerides, phospholipids, glycolipids and cardiolipins) were quickly and easily detected over a mass range spanning from ca. 200 to ca. 1600 m/z. Moreover, mass spectra with improved S/N ratio (compared to single matrices), reduced chemical noise and no formation of matrix-clusters were invariably obtained demonstrating the potential of this binary matrix to improve sensitivity.

  11. Natural products in Glycyrrhiza glabra (licorice) rhizome imaged at the cellular level by atmospheric pressure matrix-assisted laser desorption/ionization tandem mass spectrometry imaging.

    Science.gov (United States)

    Li, Bin; Bhandari, Dhaka Ram; Janfelt, Christian; Römpp, Andreas; Spengler, Bernhard

    2014-10-01

    The rhizome of Glycyrrhiza glabra (licorice) was analyzed by high-resolution mass spectrometry imaging and tandem mass spectrometry imaging. An atmospheric pressure matrix-assisted laser desorption/ionization imaging ion source was combined with an orbital trapping mass spectrometer in order to obtain high-resolution imaging in mass and space. Sections of the rhizome were imaged with a spatial resolution of 10 μm in the positive ion mode, and a large number of secondary metabolites were localized and identified based on their accurate mass and MS/MS fragmentation patterns. Major tissue-specific metabolites, including free flavonoids, flavonoid glycosides and saponins, were successfully detected and visualized in images, showing their distributions at the cellular level. The analytical power of the technique was tested in the imaging of two isobaric licorice saponins with a mass difference of only 0.02 Da. With a mass resolving power of 140 000 and a bin width of 5 ppm in the image processing, the two compounds were well resolved in full-scan mode, and appeared with different distributions in the tissue sections. The identities of the compounds and their distributions were validated in a subsequent MS/MS imaging experiment, thereby confirming their identities and excluding possible analyte interference. The use of high spatial resolution, high mass resolution and tandem mass spectrometry in imaging experiments provides significant information about the biosynthetic pathway of flavonoids and saponins in legume species, combing the spatially resolved chemical information with morphological details at the microscopic level. Furthermore, the technique offers a scheme capable of high-throughput profiling of metabolites in plant tissues.

  12. The use of matrix-assisted laser desorption ionization-time of flight mass spectrometry in the identification of Francisella tularensis

    Science.gov (United States)

    Karatuna, Onur; Çelebi, Bekir; Can, Simge; Akyar, Işın; Kiliç, Selçuk

    2016-01-01

    Francisella tularensis is the cause of the zoonotic disease tularemia and is classified among highly pathogenic bacteria (HPB) due to its low infection dose and potential for airborne transmission. In the case of HBP, there is a pressing need for rapid, accurate and reliable identification. Phenotypic identification of Francisella species is inappropriate for clinical microbiology laboratories because it is time-consuming, hazardous and subject to variable interpretation. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was recently evaluated as a useful tool for the rapid identification of a variety of microorganisms. In this study, we evaluated the use of MALDI-TOF MS for the rapid identification of Francisella tularensis and differentiation of its subspecies. Using national collection of Francisella isolates from the National Tularemia Reference Laboratory (Public Health Institution of Turkey, Ankara), a total of 75 clinical isolates were investigated by species and subspecies-specific polymerase chain reaction (PCR) test and MALDI-TOF MS. All isolates were originally identified as F. tularensis subsp. holarctica according to region of difference 1 (RD1) subspecies-specific PCR results. For all isolates MALDI-TOF MS provided results in concordance with subspecies-specific PCR analysis. Although PCR-based methods are effective in identifying Francisella species, they are labor-intensive and take longer periods of time to obtain the results when compared with MALDI-TOF MS. MALDI-TOF MS appeared to be a rapid, reliable and cost-effective identification technique for Francisella spp. Shorter analysis time and low cost make this an appealing new option in microbiology laboratories. PMID:26773181

  13. Identification of rare pathogenic bacteria in a clinical microbiology laboratory: impact of matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Seng, Piseth; Abat, Cedric; Rolain, Jean Marc; Colson, Philippe; Lagier, Jean-Christophe; Gouriet, Frédérique; Fournier, Pierre Edouard; Drancourt, Michel; La Scola, Bernard; Raoult, Didier

    2013-07-01

    During the past 5 years, matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has become a powerful tool for routine identification in many clinical laboratories. We analyzed our 11-year experience in routine identification of clinical isolates (40 months using MALDI-TOF MS and 91 months using conventional phenotypic identification [CPI]). Among the 286,842 clonal isolates, 284,899 isolates of 459 species were identified. The remaining 1,951 isolates were misidentified and required confirmation using a second phenotypic identification for 670 isolates and using a molecular technique for 1,273 isolates of 339 species. MALDI-TOF MS annually identified 112 species, i.e., 36 species/10,000 isolates, compared to 44 species, i.e., 19 species/10,000 isolates, for CPI. Only 50 isolates required second phenotypic identifications during the MALDI-TOF MS period (i.e., 4.5 reidentifications/10,000 isolates) compared with 620 isolates during the CPI period (i.e., 35.2/10,000 isolates). We identified 128 bacterial species rarely reported as human pathogens, including 48 using phenotypic techniques (22 using CPI and 37 using MALDI-TOF MS). Another 75 rare species were identified using molecular methods. MALDI-TOF MS reduced the time required for identification by 55-fold and 169-fold and the cost by 5-fold and 96-fold compared with CPI and gene sequencing, respectively. MALDI-TOF MS was a powerful tool not only for routine bacterial identification but also for identification of rare bacterial species implicated in human infectious diseases. The ability to rapidly identify bacterial species rarely described as pathogens in specific clinical specimens will help us to study the clinical burden resulting from the emergence of these species as human pathogens, and MALDI-TOF MS may be considered an alternative to molecular methods in clinical laboratories.

  14. Imaging Mass Spectrometry by Matrix-Assisted Laser Desorption/Ionization and Stress-Strain Measurements in Iontophoresis Transepithelial Corneal Collagen Cross-Linking

    Directory of Open Access Journals (Sweden)

    Paolo Vinciguerra

    2014-01-01

    Full Text Available Purpose. To compare biomechanical effect, riboflavin penetration and distribution in transepithelial corneal collagen cross-linking with iontophoresis (I-CXL, with standard cross linking (S-CXL and current transepithelial protocol (TE-CXL. Materials and Methods. The study was divided into two different sections, considering, respectively, rabbit and human cadaver corneas. In both sections corneas were divided according to imbibition protocols and irradiation power. Imaging mass spectrometry by matrix-assisted laser desorption/ionization (MALDI-IMS and stress-strain measurements were used. Forty-eight rabbit and twelve human cadaver corneas were evaluated. Results. MALDI-IMS showed a deep riboflavin penetration throughout the corneal layers with I-CXL, with a roughly lower concentration in the deepest layers when compared to S-CXL, whereas with TE-CXL penetration was considerably less. In rabbits, there was a significant increase (by 71.9% and P=0.05 in corneal rigidity after I-CXL, when compared to controls. In humans, corneal rigidity increase was not significantly different among the subgroups. Conclusions. In rabbits, I-CXL induced a significant increase in corneal stiffness as well as better riboflavin penetration when compared to controls and TE-CXL but not to S-CXL. Stress-strain in human corneas did not show significant differences among techniques, possibly because of the small sample size of groups. In conclusion, I-CXL could be a valid alternative to S-CXL for riboflavin delivery in CXL, preserving the epithelium.

  15. Coumarins as new matrices for matrix-assisted laser-desorption/ionization Fourier transform ion cyclotron resonance mass spectrometric analysis of hydrophobic compounds

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Hang, E-mail: hangwang@sjtu.edu.cn [Instrumental Analysis Center, Shanghai Jiao Tong University, Dongchuan Road 800, Shanghai 200240 (China); Dai, Bona [Instrumental Analysis Center, Shanghai Jiao Tong University, Dongchuan Road 800, Shanghai 200240 (China); Liu, Bin [Key Laboratory of Kidney Disease Pathogenesis and Intervention of Hubei Province, College of Medicine, Hubei Polytechnic University, Huangshi, Hubei 435003 (China); Lu, Han [Department of Anesthesiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine (SJTU-SM), 197, Rui Jin Er Road, Shanghai 200025 (China)

    2015-07-02

    Highlights: • Coumarins were used as new MALDI matrices. • Coumarins were used for MALDI-FT ICR MS detection of hydrophobic compounds. • DCA had improvement in detection sensitivity, stability, selectivity and reproducibility. • DCA was applied to sterols detection in yeast cells. - Abstract: Hydrophobic compounds with hydroxyl, aldehyde or ketone groups are generally difficult to detect using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), because these compounds have low proton affinity and are poorly ionized by MALDI. Herein, coumarins have been used as new matrices for MALDI-MS analysis of a variety of hydrophobic compounds with low ionization efficiency, including steroids, coenzyme Q10, a cyclic lipopeptide and cholesterol oleate. Five coumarins, including coumarin, umbelliferone, esculetin, 7-hydroxycoumarin-3-carboxylic acid (HCA) and 6,7-dihydroxycoumarin-3-carboxylic acid (DCA), were compared with the conventional matrices of 2,5-dihydroxybenzoic acid (DHB) and α-cyano-4-hydroxycinnamic acid (CHCA). Coumarins with hydroxyl or carboxylic acid groups enabled detection. Taking DCA as an example, this matrix proved to be superior to DHB or CHCA in detection sensitivity, stability, spot-to-spot and sample-to-sample reproducibility, and accuracy. DCA increased the stability of the target compounds and decreased the loss of water. The [M + Na]{sup +} peaks were observed for all target compounds by adding NaCl as an additive, and the [M − H{sub 2}O + H]{sup +} and [M + H]{sup +} peaks decreased. DCA was selected for the identification of sterols in yeast cells, and thirteen sterols were detected by Fourier transform ion cyclotron resonance (FT ICR) mass spectrometry. This work demonstrates the potential of DCA as a new matrix for detection of hydrophobic molecules by MALDI-MS and provides an alternative tool for screening sterols in antifungal research.

  16. Comparison of the Microflex LT and Vitek MS systems for routine identification of bacteria by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Martiny, Delphine; Busson, Laurent; Wybo, Ingrid; El Haj, Rachid Ait; Dediste, Anne; Vandenberg, Olivier

    2012-04-01

    This study compared the performance of three matrix-assisted laser desorption ionization-time of flight mass spectrometry systems: Microflex LT (Bruker Daltonics, Bremen, Germany), Vitek MS RUO (Axima Assurance-Saramis database; bioMérieux, Marcy l'Etoile, France), and Vitek MS IVD (bioMérieux). A total of 1,129 isolates, including 1,003 routine isolates, 73 anaerobes, and 53 bacterial enteropathogens, were tested on the Microflex LT and Axima Assurance devices. The spectra were analyzed using three databases: Biotyper (Bruker Daltonics), Saramis, and Vitek MS (bioMérieux). Among the routine isolates requiring identification to the species level (n = 986), 92.7% and 93.2% were correctly identified by the Biotyper and Vitek MS databases, respectively. The Vitek MS database is more specific for the identification of Streptococcus viridans. For the anaerobes, the Biotyper database often identified Fusobacterium isolates to only the genus level, which is of low clinical significance, whereas 20% of the Bacteroides species were not identified or were misidentified by the Vitek MS database. For the enteropathogens, the poor discrimination between Escherichia coli and Shigella explains the high proportion of unidentified organisms. In contrast to the Biotyper database, the Vitek MS database properly discriminated all of the Salmonella entrica serovar Typhi isolates (n = 5). The performance of the Saramis database was globally poorer. In conclusion, for routine procedures, the Microflex LT and Vitek-MS systems are equally good choices in terms of analytical efficiency. Other factors, including price, work flow, and lab activity, will affect the choice of a system.

  17. Chemical analysis of pharmaceuticals and explosives in fingermarks using matrix-assisted laser desorption ionization/time-of-flight mass spectrometry.

    Science.gov (United States)

    Kaplan-Sandquist, Kimberly; LeBeau, Marc A; Miller, Mark L

    2014-02-01

    Chemical analysis of latent fingermarks, "touch chemistry," has the potential of providing intelligence or forensically relevant information. Matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI/TOF MS) was used as an analytical platform for obtaining mass spectra and chemical images of target drugs and explosives in fingermark residues following conventional fingerprint development methods and MALDI matrix processing. There were two main purposes of this research: (1) develop effective laboratory methods for detecting drugs and explosives in fingermark residues and (2) determine the feasibility of detecting drugs and explosives after casual contact with pills, powders, and residues. Further, synthetic latent print reference pads were evaluated as mimics of natural fingermark residue to determine if the pads could be used for method development and quality control. The results suggest that artificial amino acid and sebaceous oil residue pads are not suitable to adequately simulate natural fingermark chemistry for MALDI/TOF MS analysis. However, the pads were useful for designing experiments and setting instrumental parameters. Based on the natural fingermark residue experiments, handling whole or broken pills did not transfer sufficient quantities of drugs to allow for definitive detection. Transferring drugs or explosives in the form of powders and residues was successful for preparing analytes for detection after contact with fingers and deposition of fingermark residue. One downfall to handling powders was that the analyte particles were easily spread beyond the original fingermark during development. Analyte particles were confined in the original fingermark when using transfer residues. The MALDI/TOF MS was able to detect procaine, pseudoephedrine, TNT, and RDX from contact residue under laboratory conditions with the integration of conventional fingerprint development methods and MALDI matrix. MALDI/TOF MS is a nondestructive

  18. A Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Method for the Identification of Anthraquinones: the Case of Historical Lakes

    Science.gov (United States)

    Sabatini, Francesca; Lluveras-Tenorio, Anna; Degano, Ilaria; Kuckova, Stepanka; Krizova, Iva; Colombini, Maria Perla

    2016-08-01

    This study deals with the identification of anthraquinoid molecular markers in standard dyes, reference lakes, and paint model systems using a micro-invasive and nondestructive technique such as matrix-assisted laser desorption/ionization time-of-flight-mass spectrometry (MALDI-ToF-MS). Red anthraquinoid lakes, such as madder lake, carmine lake, and Indian lac, have been the most widely used for painting purposes since ancient times. From an analytical point of view, identifying lakes in paint samples is challenging and developing methods that maximize the information achievable minimizing the amount of sample needed is of paramount importance. The employed method was tested on less than 0.5 mg of reference samples and required a minimal sample preparation, entailing a hydrofluoric acid extraction. The method is fast and versatile because of the possibility to re-analyze the same sample (once it has been spotted on the steel plate), testing both positive and negative modes in a few minutes. The MALDI mass spectra collected in the two analysis modes were studied and compared with LDI and simulated mass spectra in order to highlight the peculiar behavior of the anthraquinones in the MALDI process. Both ionization modes were assessed for each species. The effect of the different paint binders on dye identification was also evaluated through the analyses of paint model systems. In the end, the method was successful in detecting madder lake in archeological samples from Greek wall paintings and on an Italian funerary clay vessel, demonstrating its capabilities to identify dyes in small amount of highly degraded samples.

  19. High-Throughput Identification of Bacteria and Yeast by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry in Conventional Medical Microbiology Laboratories ▿

    Science.gov (United States)

    van Veen, S. Q.; Claas, E. C. J.; Kuijper, Ed J.

    2010-01-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is suitable for high-throughput and rapid diagnostics at low costs and can be considered an alternative for conventional biochemical and molecular identification systems in a conventional microbiological laboratory. First, we evaluated MALDI-TOF MS using 327 clinical isolates previously cultured from patient materials and identified by conventional techniques (Vitek-II, API, and biochemical tests). Discrepancies were analyzed by molecular analysis of the 16S genes. Of 327 isolates, 95.1% were identified correctly to genus level, and 85.6% were identified to species level by MALDI-TOF MS. Second, we performed a prospective validation study, including 980 clinical isolates of bacteria and yeasts. Overall performance of MALDI-TOF MS was significantly better than conventional biochemical systems for correct species identification (92.2% and 83.1%, respectively) and produced fewer incorrect genus identifications (0.1% and 1.6%, respectively). Correct species identification by MALDI-TOF MS was observed in 97.7% of Enterobacteriaceae, 92% of nonfermentative Gram-negative bacteria, 94.3% of staphylococci, 84.8% of streptococci, 84% of a miscellaneous group (mainly Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella [HACEK]), and 85.2% of yeasts. MALDI-TOF MS had significantly better performance than conventional methods for species identification of staphylococci and genus identification of bacteria belonging to HACEK group. Misidentifications by MALDI-TOF MS were clearly associated with an absence of sufficient spectra from suitable reference strains in the MALDI-TOF MS database. We conclude that MALDI-TOF MS can be implemented easily for routine identification of bacteria (except for pneumococci and viridans streptococci) and yeasts in a medical microbiological laboratory. PMID:20053859

  20. Direct demonstration of tissue uptake of an inhaled drug: proof-of-principle study using matrix-assisted laser desorption ionization mass spectrometry imaging.

    Science.gov (United States)

    Fehniger, Thomas E; Végvári, Akos; Rezeli, Melinda; Prikk, Kaiu; Ross, Peeter; Dahlbäck, Magnus; Edula, Goutham; Sepper, Ruth; Marko-Varga, György

    2011-11-01

    Drug therapy is often directed to specific organ and tissue compartments where the mode of action of the compound affects specifically targeted biological processes. However, the direct measurement of drug uptake in terms of a time kinetic and concentrations attained at the local sites has not been readily available as a clinical index for most drugs. A proof-of-principle study was conducted to test the utility of applying matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) to demonstrate the qualitative distribution pattern of a locally administered drug within tissue sites of targeted action. Here we have measured the occurrence of an inhaled bronchodilator, the muscarinic receptor antagonist ipratropium, within human bronchial biopsies obtained by fiber optic bronchoscopy shortly after dosing exposure. Cryo-preserved biopsy samples from five subjects being evaluated for airway obstruction or potential tumor development were prepared as thin frozen sections. Samples coated with a MALDI matrix were analyzed by a MALDI LTQ Orbitrap XL mass spectrometer at large (100 μm) and small (30 μm) raster sizes. Our results demonstrate that ipratropium is rapidly absorbed into the airway wall. Ipratropium parent ion (m/z 332.332) and daughter ions (m/z 166.2 and 290.2) were coincidently partitioned within submucosal spaces containing targeted airway smooth muscle in four out of five subjects. The signal intensity of ipratropium fragment ions provided estimates that local drug concentrations between 3 and 80 nM were achieved within the airway wall. To our knowledge, this is the first reported study in applying MALDI-MSI to demonstrate the localization of a drug administered at therapeutic levels. The study highlights the potential benefit of MALDI-MSI to provide important measurements of drug efficacy in clinical settings.

  1. Use of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of molds of the Fusarium genus.

    Science.gov (United States)

    Triest, David; Stubbe, Dirk; De Cremer, Koen; Piérard, Denis; Normand, Anne-Cécile; Piarroux, Renaud; Detandt, Monique; Hendrickx, Marijke

    2015-02-01

    The rates of infection with Fusarium molds are increasing, and a diverse number of Fusarium spp. belonging to different species complexes can cause infection. Conventional species identification in the clinical laboratory is time-consuming and prone to errors. We therefore evaluated whether matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a useful alternative. The 289 Fusarium strains from the Belgian Coordinated Collections of Microorganisms (BCCM)/Institute of Hygiene and Epidemiology Mycology (IHEM) culture collection with validated sequence-based identities and comprising 40 species were used in this study. An identification strategy was developed, applying a standardized MALDI-TOF MS assay and an in-house reference spectrum database. In vitro antifungal testing was performed to assess important differences in susceptibility between clinically relevant species/species complexes. We observed that no incorrect species complex identifications were made by MALDI-TOF MS, and 82.8% of the identifications were correct to the species level. This success rate was increased to 91% by lowering the cutoff for identification. Although the identification of the correct species complex member was not always guaranteed, antifungal susceptibility testing showed that discriminating between Fusarium species complexes can be important for treatment but is not necessarily required between members of a species complex. With this perspective, some Fusarium species complexes with closely related members can be considered as a whole, increasing the success rate of correct identifications to 97%. The application of our user-friendly MALDI-TOF MS identification approach resulted in a dramatic improvement in both time and accuracy compared to identification with the conventional method. A proof of principle of our MALDI-TOF MS approach in the clinical setting using recently isolated Fusarium strains demonstrated its validity.

  2. Liquid chromatography electrospray ionization and matrix-assisted laser desorption ionization tandem mass spectrometry for the analysis of lipid raft proteome of monocytes

    Energy Technology Data Exchange (ETDEWEB)

    Zhang Nan [Department of Chemistry, University of Alberta, Edmonton, Alberta (Canada); Shaw, Andrew R.E. [Department of Chemistry, University of Alberta, Edmonton, Alberta (Canada)], E-mail: andrewsh@cancerboard.ab.ca; Li Nan; Chen Rui [Department of Chemistry, University of Alberta, Edmonton, Alberta (Canada); Mak, Allan; Hu Xiuying [Department of Oncology, University of Alberta, Edmonton, Alberta (Canada); Young, Nelson; Wishart, David [Department of Computing Science, University of Alberta, Edmonton, Alberta (Canada); Li Liang, E-mail: Liang.Li@ualberta.ca

    2008-10-03

    Lipid rafts are dynamic assemblies of cholesterol and glycolipid that form detergent-insoluble microdomains within membrane lipid bilayers. Because rafts can be separated by flotation on sucrose gradients, interrogation by mass spectrometry (MS) provides a valuable new insight into lipid raft function. Here we combine liquid chromatography (LC) electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI) MS/MS to corroborate and extend our previous description of lipid raft proteomes derived from the monocytic cell line THP-1. Interestingly, LC-ESI and MALDI MS/MS identify largely non-overlapping, and therefore, potentially complementary protein populations. Using the combined approach, we detected 277 proteins compared to 52 proteins obtained with the original gel-based MALDI MS. We confirmed the presence of 47 of the original 52 proteins demonstrating the consistency of the lipid raft preparations. We demonstrated by immunoblotting that Rac 1 and Rac 2, two of the 52 proteins we failed to confirm, were indeed absent from the lipid raft fractions. The majority of new proteins were cytoskeletal proteins and their regulators, proteins implicated in membrane fusion and vesicular trafficking or signaling molecules. Our results therefore, confirm and extend previous evidence indicating lipid rafts of monocytic cells are specialized for cytoskeletal assembly and vesicle trafficking. Of particular interest, we detected SNAP-23, basigin, Glut-4 and pantophysin in lipid rafts. Since these proteins are implicated in both vesicular trafficking and gamete fusion, lipid rafts may play a common role in these processes. It is evident that the combination of LC-ESI and LC-MALDI MS/MS increases the proteome coverage which allows better understanding of the lipid raft function.

  3. 4-Chloro-α-cyanocinnamic acid is an efficient soft matrix for cyanocobalamin detection in foodstuffs by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS).

    Science.gov (United States)

    Calvano, Cosima Damiana; Ventura, Giovanni; Palmisano, Francesco; Cataldi, Tommaso R I

    2016-09-01

    4-Chloro-α-cyanocinnamic acid (ClCCA) is a very useful matrix able to give the protonated adduct [M+H](+) of intact cyanocobalamin (CNCbl) as the base peak (m/z 1355.58) in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS). The only fragment observed is [M-CN + H](+•) formed through the facile (•) CN neutral loss reflecting the fairly low Co-C bond energy. All other investigated proton transfer matrices, including α-cyano-4-hydroxycinnamic acid, para-nitroaniline and 2,5-dihydroxybenzoic acid, give rise to a complete decyanation of CNCbl with concomitant formation of [M-CN + H](+•) , [M-CN + Na](+•) and [M-CN + K](+•) adducts at m/z 1329.57, 1351.55 and 1367.51, respectively. Depending on the matrix used, a variable degree of fragmentation involving the α-side axial ligand was observed. A plausible explanation of the specific behaviour of 4-chloro-α-cyanocinnamic acid as a soft matrix is discussed. Tandem mass spectra of both [M + H](+) and [M-CN + H](+•) ions were obtained and product ions successfully assigned. The possibility of detecting the protonated adduct of intact CNCbl was exploited in foodstuff samples such as cow milk and hen egg yolk by MALDI tandem MS upon sample extraction. We believe that our data provide strong basis for the application of MALDI tandem MS in the qualitative analysis of natural CNCbl, including fish, liver and meat samples. Copyright © 2016 John Wiley & Sons, Ltd.

  4. Analysis of methicillin-resistant Staphylococcus aureus major clonal lineages by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS).

    Science.gov (United States)

    Zhang, Tingting; Ding, Jinya; Rao, Xiancai; Yu, Jingbo; Chu, Meiling; Ren, Wei; Wang, Lu; Xue, Wencheng

    2015-10-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen associated with nosocomial infections in many countries. Multilocus sequence typing (MLST) is one of the genetic typing methods used to type MRSA with a high discriminatory power, however, it is labor-intensive, timely, and costly. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) coupled with ClinProTools is a potential tool to discover biomarker peaks and to generate a classification model based on highly sophisticated mathematical algorithms to discriminate clonal lineages. We investigated the performance of MALDI-TOF MS for discriminating 154 MRSA-ST239, 72 MRSA-ST5, 30 MRSA-ST59, 14 MRSA-ST45, and 20 MRSA-OST (other clonal lineages). Our results indicate that the model construction and validation have good potency to discriminate ST45 from other lineages with a sensitivity and a specificity of both 100%, and a sensitivity of 95.80% and a specificity of 94.62% to identify ST239. For Biotyper classification, the sensitivity and specificity were more than of 90% for ST239, ST59 and ST45, whereas only 81.94% sensitivity for ST5. By single-peak analysis, the peaks m/z 4808 and 9614 can correctly discriminate ST45 a sensitivity and a specificity of both 100%; the peak m/z 6554 can also discriminate ST239 with a sensitivity of 91.9% and a specificity of 85.4%. In conclusion, MALDI-TOF MS coupled with ClinProTools has a high detection performance for MRSA typing with obvious advantages of being rapid, highly accurate, and being a low cost in comparison with MLST.

  5. Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry for species identification of Acinetobacter strains isolated from blood cultures.

    Science.gov (United States)

    Kishii, K; Kikuchi, K; Matsuda, N; Yoshida, A; Okuzumi, K; Uetera, Y; Yasuhara, H; Moriya, K

    2014-05-01

    The clinical relevance of Acinetobacter species, other than A. baumannii, as human pathogens has not been sufficiently assessed owing to the insufficiency of simple phenotypic clinical diagnostic laboratory tests. Infections caused by these organisms have different impacts on clinical outcome and require different treatment and management approaches. It is therefore important to correctly identify Acinetobacter species. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been introduced to identify a wide range of microorganisms in clinical laboratories, but only a few studies have examined its utility for identifying Acinetobacter species, particularly those of the non-Acinetobacter baumannii complex. We therefore evaluated MALDI-TOF MS for identification of Acinetobacter species by comparing it with sequence analysis of rpoB using 123 isolates of Acinetobacter species from blood. Of the isolates examined, we identified 106/123 (86.2%) to species, and 16/123 (13.0%) could only be identified as acinetobacters. The identity of one isolate could not be established. Of the 106 species identified, 89/106 (84.0%) were confirmed by rpoB sequence analysis, and 17/106 (16.0%) were discordant. These data indicate correct identification of 89/123 (72.4%) isolates. Surprisingly, all blood culture isolates were identified as 13 species of Acinetobacter, and the incidence of Acinetobacter pittii was unexpectedly high (42/123; 34.1%) and exceeded that of A. baumannii (22/123; 17.9%). Although the present identification rate using MALDI-TOF MS is not acceptable for species-level identification of Acinetobacter, further expansion of the database should remedy this situation.

  6. A Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Method for the Identification of Anthraquinones: the Case of Historical Lakes

    Science.gov (United States)

    Sabatini, Francesca; Lluveras-Tenorio, Anna; Degano, Ilaria; Kuckova, Stepanka; Krizova, Iva; Colombini, Maria Perla

    2016-11-01

    This study deals with the identification of anthraquinoid molecular markers in standard dyes, reference lakes, and paint model systems using a micro-invasive and nondestructive technique such as matrix-assisted laser desorption/ionization time-of-flight-mass spectrometry (MALDI-ToF-MS). Red anthraquinoid lakes, such as madder lake, carmine lake, and Indian lac, have been the most widely used for painting purposes since ancient times. From an analytical point of view, identifying lakes in paint samples is challenging and developing methods that maximize the information achievable minimizing the amount of sample needed is of paramount importance. The employed method was tested on less than 0.5 mg of reference samples and required a minimal sample preparation, entailing a hydrofluoric acid extraction. The method is fast and versatile because of the possibility to re-analyze the same sample (once it has been spotted on the steel plate), testing both positive and negative modes in a few minutes. The MALDI mass spectra collected in the two analysis modes were studied and compared with LDI and simulated mass spectra in order to highlight the peculiar behavior of the anthraquinones in the MALDI process. Both ionization modes were assessed for each species. The effect of the different paint binders on dye identification was also evaluated through the analyses of paint model systems. In the end, the method was successful in detecting madder lake in archeological samples from Greek wall paintings and on an Italian funerary clay vessel, demonstrating its capabilities to identify dyes in small amount of highly degraded samples.

  7. Rapid Identification of microbes in positive blood cultures by use of the vitek MS matrix-assisted laser desorption ionization-time of flight mass spectrometry system.

    Science.gov (United States)

    Foster, Arnold G W

    2013-11-01

    Sepsis is a major cause of death worldwide among nonhospitalized people and hospitalized patients. A wide range of pathogens are involved, and the correct identification and correct antimicrobial therapy are critical to ensure optimal clinical outcomes. With the recent introduction of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), rapid identification of bacteria and fungi is now possible. The purpose of this study was to develop a rapid technique for identifying organisms in positive blood cultures using the Vitek MS system (bioMérieux). This technique is a lysis centrifugation method which involves a four-step washing and centrifugation procedure. A total of 253 positive monomicrobial blood cultures (Bactec Plus aerobic, anaerobic, and pediatric bottles) were tested using the Vitek MS system (KnowledgeBase version 2.0), with 92.1% and 88.1% of organisms overall being identified to the genus level and the species level, respectively. Of 161 Gram-positive bacterial isolates, 95.7% and 90.1% were identified to the genus level and the species level, respectively; of 92 Gram-negative bacterial isolates, 84.7% and 83.7% were identified to the genus level and the species level, respectively. The results obtained using this method demonstrate that the Vitek MS system can be used for rapid and effective identification of bacteria from positive blood cultures within 30 to 45 min after the positive signal has been provided by the Bactec FX blood culture system (Becton, Dickinson). This will lead to faster administration of the appropriate antimicrobial therapy and increase the chances for optimal clinical outcomes for patients.

  8. Rapid detection of enterobacteriaceae producing extended spectrum beta-lactamases directly from positive blood cultures by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Oviaño, M; Fernández, B; Fernández, A; Barba, M J; Mouriño, C; Bou, G

    2014-11-01

    Bacteria that produce extended-spectrum β-lactamases (ESBLs) are an increasing healthcare problem and their rapid detection is a challenge that must be overcome in order to optimize antimicrobial treatment and patient care. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has been used to determine resistance to β-lactams, including carbapenems in Enterobacteriaceae, but the methodology has not been fully validated as it remains time-consuming. We aimed to assess whether MALDI-TOF can be used to detect ESBL-producing Enterobacteriaceae from positive blood culture bottles in clinical practice. In the assay, 141 blood cultures were tested, 13 of them were real bacteraemias and 128 corresponded to blood culture bottles seeded with bacterial clinical isolates. Bacteraemias were analysed by MALDI-TOF after a positive growth result and the 128 remaining blood cultures 24 h after the bacterial seeding. β-lactamase activity was determined through the profile of peaks associated with the antibiotics cefotaxime and ceftazidime and its hydrolyzed forms. Clavulanic acid was added to rule out the presence of non-ESBL mechanisms. Overall data show a 99% (103 out of 104) sensitivity in detecting ESBL in positive blood cultures. Data were obtained in 90 min (maximum 150 min). The proposed methodology has a great impact on the early detection of ESBL-producing Enterobacteriaceae from positive blood cultures, being a rapid and efficient method and allowing early administration of an appropriate antibiotic therapy.

  9. Interfacing droplet microfluidics with matrix-assisted laser desorption/ionization mass spectrometry: label-free content analysis of single droplets.

    Science.gov (United States)

    Küster, Simon K; Fagerer, Stephan R; Verboket, Pascal E; Eyer, Klaus; Jefimovs, Konstantins; Zenobi, Renato; Dittrich, Petra S

    2013-02-05

    Droplet-based microfluidic systems have become a very powerful tool to miniaturize chemical and biological reactions. However, droplet content analysis remains challenging and relies almost exclusively on optical methods such as fluorescence spectroscopy. Hence, labeling of the analyte is typically required which impedes a more universal applicability of microdroplets. Here we present a novel interface coupling droplet microfluidics and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry for label-free content analysis of single droplets. Nanoliter aqueous droplets immersed in perfluorinated oil are created in a microfluidic T-junction, transferred into a capillary, and deposited on a high-density microarray MALDI plate mounted on a motorized xy-stage. The fully automated system is robust and reliable due to two unique features. First, a simple optical droplet detection system is used to synchronize stage movement and exit of droplets from the capillary. Second, the microarray plate contains an array of over 26,000 hydrophilic spots within a hydrophobic coating, each spot acting as a recipient to confine the droplets and to prevent cross-contamination. The MALDI matrix can also be applied using our system by spotting matrix droplets on the microarray in a separate run. To demonstrate the potential of our system, we studied the enzymatic cleavage of angiotensin I by angiotensin converting enzyme and monitored the increasing concentration of the product angiotensin II over time. The interface provides a robust and fully automated method for rapid label-free and information-rich content analysis of single droplets. With the high number of droplets per plate, this method is particularly suitable for high-throughput screening applications.

  10. Rapid identification of Bacillus anthracis spores in suspicious powder samples by using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS).

    Science.gov (United States)

    Dybwad, Marius; van der Laaken, Anton L; Blatny, Janet Martha; Paauw, Armand

    2013-09-01

    Rapid and reliable identification of Bacillus anthracis spores in suspicious powders is important to mitigate the safety risks and economic burdens associated with such incidents. The aim of this study was to develop and validate a rapid and reliable laboratory-based matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis method for identifying B. anthracis spores in suspicious powder samples. A reference library containing 22 different Bacillus sp. strains or hoax materials was constructed and coupled with a novel classification algorithm and standardized processing protocol for various powder samples. The method's limit of B. anthracis detection was determined to be 2.5 × 10(6) spores, equivalent to a 55-μg sample size of the crudest B. anthracis-containing powder discovered during the 2001 Amerithrax incidents. The end-to-end analysis method was able to successfully discriminate among samples containing B. anthracis spores, closely related Bacillus sp. spores, and commonly encountered hoax materials. No false-positive or -negative classifications of B. anthracis spores were observed, even when the analysis method was challenged with a wide range of other bacterial agents. The robustness of the method was demonstrated by analyzing samples (i) at an external facility using a different MALDI-TOF MS instrument, (ii) using an untrained operator, and (iii) using mixtures of Bacillus sp. spores and hoax materials. Taken together, the observed performance of the analysis method developed demonstrates its potential applicability as a rapid, specific, sensitive, robust, and cost-effective laboratory-based analysis tool for resolving incidents involving suspicious powders in less than 30 min.

  11. High-throughput identification of bacteria and yeast by matrix-assisted laser desorption ionization-time of flight mass spectrometry in conventional medical microbiology laboratories.

    Science.gov (United States)

    van Veen, S Q; Claas, E C J; Kuijper, Ed J

    2010-03-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is suitable for high-throughput and rapid diagnostics at low costs and can be considered an alternative for conventional biochemical and molecular identification systems in a conventional microbiological laboratory. First, we evaluated MALDI-TOF MS using 327 clinical isolates previously cultured from patient materials and identified by conventional techniques (Vitek-II, API, and biochemical tests). Discrepancies were analyzed by molecular analysis of the 16S genes. Of 327 isolates, 95.1% were identified correctly to genus level, and 85.6% were identified to species level by MALDI-TOF MS. Second, we performed a prospective validation study, including 980 clinical isolates of bacteria and yeasts. Overall performance of MALDI-TOF MS was significantly better than conventional biochemical systems for correct species identification (92.2% and 83.1%, respectively) and produced fewer incorrect genus identifications (0.1% and 1.6%, respectively). Correct species identification by MALDI-TOF MS was observed in 97.7% of Enterobacteriaceae, 92% of nonfermentative Gram-negative bacteria, 94.3% of staphylococci, 84.8% of streptococci, 84% of a miscellaneous group (mainly Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella [HACEK]), and 85.2% of yeasts. MALDI-TOF MS had significantly better performance than conventional methods for species identification of staphylococci and genus identification of bacteria belonging to HACEK group. Misidentifications by MALDI-TOF MS were clearly associated with an absence of sufficient spectra from suitable reference strains in the MALDI-TOF MS database. We conclude that MALDI-TOF MS can be implemented easily for routine identification of bacteria (except for pneumococci and viridans streptococci) and yeasts in a medical microbiological laboratory.

  12. Performances of the Vitek MS matrix-assisted laser desorption ionization-time of flight mass spectrometry system for rapid identification of bacteria in routine clinical microbiology.

    Science.gov (United States)

    Dubois, Damien; Grare, Marion; Prere, Marie-Françoise; Segonds, Christine; Marty, Nicole; Oswald, Eric

    2012-08-01

    Rapid and cost-effective matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based systems will replace conventional phenotypic methods for routine identification of bacteria. We report here the first evaluation of the new MALDI-TOF MS-based Vitek MS system in a large clinical microbiology laboratory. This system uses an original spectrum classifier algorithm and a specific database designed for the identification of clinically relevant species. We have tested 767 routine clinical isolates representative of 50 genera and 124 species. Vitek MS-based identifications were performed by means of a single deposit on a MALDI disposable target without any prior extraction step and compared with reference identifications obtained mainly with the VITEK2 phenotypic system; if the identifications were discordant, molecular techniques provided reference identifications. The Vitek MS system provided 96.2% correct identifications to the species level (86.7%), to the genus level (8.2%), or within a range of species belonging to different genera (1.3%). Conversely, 1.3% of isolates were misidentified and 2.5% were unidentified, partly because the species was not included in the database; a second deposit provided a successful identification for 0.8% of isolates unidentified with the first deposit. The Vitek MS system is a simple, convenient, and accurate method for routine bacterial identification with a single deposit, considering the high bacterial diversity studied and as evidenced by the low prevalence of species without correct identification. In addition to a second deposit in uncommon cases, expanding the spectral database is expected to further enhance performances.

  13. Identification of Haemophilus influenzae Type b Isolates by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

    Science.gov (United States)

    Månsson, Viktor; Kostrzewa, Markus; Nilson, Bo; Riesbeck, Kristian

    2015-01-01

    Haemophilus influenzae type b (Hib) is, in contrast to non-type b H. influenzae, associated with severe invasive disease, such as meningitis and epiglottitis, in small children. To date, accurate H. influenzae capsule typing requires PCR, a time-consuming and cumbersome method. Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) provides rapid bacterial diagnostics and is increasingly used in clinical microbiology laboratories. Here, MALDI-TOF MS was evaluated as a novel approach to separate Hib from other H. influenzae. PCR-verified Hib and non-Hib reference isolates were selected based on genetic and spectral characteristics. Mass spectra of reference isolates were acquired and used to generate different classification algorithms for Hib/non-Hib differentiation using both ClinProTools and the MALDI Biotyper software. A test series of mass spectra from 33 Hib and 77 non-Hib isolates, all characterized by PCR, was used to evaluate the algorithms. Several algorithms yielded good results, but the two best were a ClinProTools model based on 22 separating peaks and subtyping main spectra (MSPs) using MALDI Biotyper. The ClinProTools model had a sensitivity of 100% and a specificity of 99%, and the results were 98% reproducible using a different MALDI-TOF MS instrument. The Biotyper subtyping MSPs had a sensitivity of 97%, a specificity of 100%, and 93% reproducibility. Our results suggest that it is possible to use MALDI-TOF MS to differentiate Hib from other H. influenzae. This is a promising method for rapidly identifying Hib in unvaccinated populations and for the screening and surveillance of Hib carriage in vaccinated populations. PMID:25926500

  14. Identification of Haemophilus influenzae Type b Isolates by Use of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

    Science.gov (United States)

    Månsson, Viktor; Resman, Fredrik; Kostrzewa, Markus; Nilson, Bo; Riesbeck, Kristian

    2015-07-01

    Haemophilus influenzae type b (Hib) is, in contrast to non-type b H. influenzae, associated with severe invasive disease, such as meningitis and epiglottitis, in small children. To date, accurate H. influenzae capsule typing requires PCR, a time-consuming and cumbersome method. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) provides rapid bacterial diagnostics and is increasingly used in clinical microbiology laboratories. Here, MALDI-TOF MS was evaluated as a novel approach to separate Hib from other H. influenzae. PCR-verified Hib and non-Hib reference isolates were selected based on genetic and spectral characteristics. Mass spectra of reference isolates were acquired and used to generate different classification algorithms for Hib/non-Hib differentiation using both ClinProTools and the MALDI Biotyper software. A test series of mass spectra from 33 Hib and 77 non-Hib isolates, all characterized by PCR, was used to evaluate the algorithms. Several algorithms yielded good results, but the two best were a ClinProTools model based on 22 separating peaks and subtyping main spectra (MSPs) using MALDI Biotyper. The ClinProTools model had a sensitivity of 100% and a specificity of 99%, and the results were 98% reproducible using a different MALDI-TOF MS instrument. The Biotyper subtyping MSPs had a sensitivity of 97%, a specificity of 100%, and 93% reproducibility. Our results suggest that it is possible to use MALDI-TOF MS to differentiate Hib from other H. influenzae. This is a promising method for rapidly identifying Hib in unvaccinated populations and for the screening and surveillance of Hib carriage in vaccinated populations.

  15. Rapid identification of Burkholderia mallei and Burkholderia pseudomallei by intact cell Matrix-assisted Laser Desorption/Ionisation mass spectrometric typing

    Directory of Open Access Journals (Sweden)

    Karger Axel

    2012-10-01

    Full Text Available Abstract Background Burkholderia (B. pseudomallei and B. mallei are genetically closely related species. B. pseudomallei causes melioidosis in humans and animals, whereas B. mallei is the causative agent of glanders in equines and rarely also in humans. Both agents have been classified by the CDC as priority category B biological agents. Rapid identification is crucial, because both agents are intrinsically resistant to many antibiotics. Matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-TOF MS has the potential of rapid and reliable identification of pathogens, but is limited by the availability of a database containing validated reference spectra. The aim of this study was to evaluate the use of MALDI-TOF MS for the rapid and reliable identification and differentiation of B. pseudomallei and B. mallei and to build up a reliable reference database for both organisms. Results A collection of ten B. pseudomallei and seventeen B. mallei strains was used to generate a library of reference spectra. Samples of both species could be identified by MALDI-TOF MS, if a dedicated subset of the reference spectra library was used. In comparison with samples representing B. mallei, higher genetic diversity among B. pseudomallei was reflected in the higher average Eucledian distances between the mass spectra and a broader range of identification score values obtained with commercial software for the identification of microorganisms. The type strain of B. pseudomallei (ATCC 23343 was isolated decades ago and is outstanding in the spectrum-based dendrograms probably due to massive methylations as indicated by two intensive series of mass increments of 14 Da specifically and reproducibly found in the spectra of this strain. Conclusions Handling of pathogens under BSL 3 conditions is dangerous and cumbersome but can be minimized by inactivation of bacteria with ethanol, subsequent protein extraction under BSL 1 conditions and MALDI-TOF MS

  16. Matrix-assisted laser desorption ionization-time of flight mass spectrometry identification of yeasts is contingent on robust reference spectra.

    Directory of Open Access Journals (Sweden)

    Angie Pinto

    Full Text Available BACKGROUND: Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS for yeast identification is limited by the requirement for protein extraction and for robust reference spectra across yeast species in databases. We evaluated its ability to identify a range of yeasts in comparison with phenotypic methods. METHODS: MALDI-TOF MS was performed on 30 reference and 167 clinical isolates followed by prospective examination of 67 clinical strains in parallel with biochemical testing (total n = 264. Discordant/unreliable identifications were resolved by sequencing of the internal transcribed spacer region of the rRNA gene cluster. PRINCIPAL FINDINGS: Twenty (67%; 16 species, and 24 (80% of 30 reference strains were identified to species, (spectral score ≥2.0 and genus (score ≥1.70-level, respectively. Of clinical isolates, 140/167 (84% strains were correctly identified with scores of ≥2.0 and 160/167 (96% with scores of ≥1.70; amongst Candida spp. (n = 148, correct species assignment at scores of ≥2.0, and ≥1.70 was obtained for 86% and 96% isolates, respectively (vs. 76.4% by biochemical methods. Prospectively, species-level identification was achieved for 79% of isolates, whilst 91% and 94% of strains yielded scores of ≥1.90 and ≥1.70, respectively (100% isolates identified by biochemical methods. All test scores of 1.70-1.90 provided correct species assignment despite being identified to "genus-level". MALDI-TOF MS identified uncommon Candida spp., differentiated Candida parapsilosis from C. orthopsilosis and C. metapsilosis and distinguished between C. glabrata, C. nivariensis and C. bracarensis. Yeasts with scores of <1.70 were rare species such as C. nivariensis (3/10 strains and C. bracarensis (n = 1 but included 4/12 Cryptococcus neoformans. There were no misidentifications. Four novel species-specific spectra were obtained. Protein extraction was essential for reliable results

  17. Teaching Microbial Identification with Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS and Bioinformatics Tools

    Directory of Open Access Journals (Sweden)

    Wenfa Ng

    2013-01-01

    Full Text Available Ever since the first observation of “animalcules” under a microscope, and the subsequent discovery of microorganisms of myriad size, shape, pigmentation and motility modes, classification in aid of microbial identification is key to understanding inter-relationships between diverse microbes. Combining universal applicability with robustness, 16S rRNA sequencing is the gold standard for microbial typing; however, recent developments in clinical diagnostics have called attention to a shift towards PCR-independent instrumentation and methods given PCR’s requirement for expensive and complex sample preparation. Using ribosomal proteins as biomarkers for evolutionary relatedness, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS - originally developed for the soft ionization of proteins and peptides in proteomics studies - has been successfully applied to identifying bacteria, archaea, fungi and viruses to the species, and, on occasions, sub-species level. Though experimentally proven and increasingly adopted in the clinic, the relatively low-cost (on a per sample basis and rapid MALDI-TOF MS microbial identification technique, along with its theoretical principles and methodology, is a conspicuous absentee in contemporary microbiology curricula. Motivated by a desire to close the curriculum gap, this article describes a discovery-based activity for teaching microbial identification - using MALDI-TOF MS in combination with open-source genomics and proteomics search tools – while providing tips on mass spectra interpretation and activity implementation for lowering the barrier for classroom adoption. Infused with inquiry-based learning concepts guiding students in identifying microbes from environmental water samples with unknown species diversity, the activity spurs students’ learning by igniting their spirit of inquiry, which leads to better mastery of concepts; a significant departure from

  18. Selective reduction of C=C double bonds in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of microcystins.

    Science.gov (United States)

    Deleuze, Christelle; De Pauw, Edwin; Quinton, Loic

    2010-01-01

    Cyanobacteria are photosynthetic bacteria encountered in various aquatic environments. Some of them are able to produce powerful toxins called cyanotoxins. Among cyanotoxins, microcystins (MCs) constitute a group of closely related cyclic heptapeptides. Their sequences are made up of classical amino acids as well as post- translational modified ones. Interestingly, in vivo metabolism of microcystins seems to be greatly dependent on various minor structural differences and particularly those of the seventh amino acid, which can be either dehydroalanine (or a derivative), dehydroaminobutyric acid (or a derivative), serine or alanine. As a consequence, microcystins have been classified on the basis of the nature of this singular amino acid. A major difficulty in the classification of such toxins is that some of them share the same molecular masses and the same molecular formulas. Consequently, a simple mass measurement is not sufficient to determine the structure and the class of a toxin of interest. Heavy and expensive techniques are used to classify them, such as multi-dimensional nuclear magnetic resonance and amino acid analysis. In this work, a new matrix-assisted laser desorption/ionization time-of-flight method leading to an easy classification of MCs is proposed. The methodology relies on the reductive properties of the matrix 1,5-diaminonaphtalene (1,5-DAN) which appears to be able to selectively reduce the double carbon-carbon bond belonging to the seventh amino acid. Moreover, the yield of reduction seems to be influenced by the degree of substitution of this double bond, allowing a discrimination between dehydroalanine and dehydroaminobutyric acid. This selective reduction was confirmed by the study of three synthetic peptides by mass spectrometry and tandem mass spectrometry. According to these results, the use of reductive matrices seems to be promising in the study of microcystins and in their classification. More generally, 1,5-DAN allows the selective

  19. Determination of the fatty acid composition of saponified vegetable oils using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Ayorinde, F O; Garvin, K; Saeed, K

    2000-01-01

    A method using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) for the determination of the fatty acid composition of vegetable oils is described and illustrated with the analysis of palm kernel oil, palm oil, olive oil, canola oil, soybean oil, vernonia oil, and castor oil. Solutions of the saponified oils, mixed with the matrix, meso-tetrakis(pentafluorophenyl)porphyrin, provided reproducible MALDI-TOF spectra in which the ions were dominated by sodiated sodium carboxylates [RCOONa + Na]+. Thus, palm kernel oil was found to contain capric acid, lauric acid, myristic acid, palmitic acid, oleic acid, and stearic acid. Palm oil had a fatty acid profile including palmitic, linoleic, oleic, and stearic. The relative percentages of the fatty acids in olive oil were palmitoleic (1.2 +/- 0.5), palmitic (10.9 +/- 0.8), linoleic (0.6 +/- 0.1), linoleic (16.5 +/- 0.8), and oleic (70.5 +/- 1.2). For soybean oil, the relative percentages were: palmitoleic (0.4 +/- 0.4), palmitic (6.0 +/- 1.3), linolenic (14.5 +/- 1.8), linoleic (50.1 +/- 4.0), oleic (26.1 +/- 1.2), and stearic (2.2 +/- 0.7). This method was also applied to the analysis of two commercial soap formulations. The first soap gave a fatty acid profile that included: lauric (19.4% +/- 0.8), myristic (9.6% +/- 0.5), palmitoleic (1.9% +/- 0.3), palmitic (16.3% +/- 0.9), linoleic (5.6% +/- 0.4), oleic (37.1% +/- 0.8), and stearic (10.1% +/- 0.7) and that of the second soap was: lauric (9.3% +/- 0.3), myristic (3.8% +/- 0.5), palmitoleic (3.1% +/- 0.8), palmitic (19.4% +/- 0.8), linoleic (4.9% +/- 0.7), oleic (49.5% +/- 1.1), and stearic (10.0% +/- 0.9). The MALDI-TOFMS method described in this communication is simpler and less time-consuming than the established transesterification method that is coupled with analysis by gas chromatography/mass spectrometry (GC/MS). The new method could be used routinely to determine the qualitative fatty acid composition of vegetable oils

  20. Matrix-assisted laser desorption ionization time-of-flight mass spectrometric analysis of degradation products after treatment of methylene blue aqueous solution with three-dimensionally integrated microsolution plasma

    Science.gov (United States)

    Shirafuji, Tatsuru; Nomura, Ayano; Hayashi, Yui; Tanaka, Kenji; Goto, Motonobu

    2016-01-01

    Methylene blue can be degraded in three-dimensionally integrated microsolution plasma. The degradation products have been analyzed by matrix-assisted laser desorption ionization time-of-flight (MALDI TOF) mass spectrometry to understand the degradation mechanisms. The results of MALDI TOF mass spectrometry have shown that sulfoxide is formed at the first stage of the oxidation. Then, partial oxidation proceeds on the methyl groups left on the sulfoxide. The sulfoxide is subsequently separated to two benzene derivatives. Finally, weak functional groups are removed from the benzene derivatives.

  1. Fragmentation study of rutin, a naturally occurring flavone glycoside cationized with different alkali metal ions, using post-source decay matrix-assisted laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Kéki, S; Deák, G; Zsuga, M

    2001-12-01

    A post-source decay matrix-assisted laser desorption/ionization mass spectrometric (PSD-MALDI-MS) study of rutin, a naturally occurring flavone glycoside cationized with different alkali metal ions, is reported. The fragmentations of rutin were performed by selecting the [R + Cat]+ peaks for PSD, where R represents a rutin molecule and Cat an alkali metal ion (Li+, Na+, K+). The PSD-MALDI mass spectra showed, depending on Cat, different fragmentation patterns with respect to both the quality and quantity of the fragment ions formed. The intensity of fragmentation decreased in the order Li+ > Na+ > K+. The fragmentation mechanism and an explanation for the observed differences are suggested.

  2. Using Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to analyze differentially expressed brain polypeptides in scrapie strain 22L-infected BALB/c mice

    Institute of Scientific and Technical Information of China (English)

    Xiangyu Liao; Chuanjing Ju; Jiayu Wan; Wensen Liu; Xin Tang; Wufei Zhu; Na Xu; Jing Xu; Nan Li; Yaping Chang

    2011-01-01

    Differentially expressed polypeptides in the brain of a BALB/c mouse model infected with scrapie strain 22L were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The results showed that 21 peptides were down-regulated, with peptides of mass-to-charge ratio 758.772 5 and mass-to-charge ratio 5 432.206 9, demonstrating the most significant decreases. These finding suggest that these peptides are candidate biomarkers and may play an important role in the pathogenesis of prion disease.

  3. Characterization of Achromobacter Species in Cystic Fibrosis Patients: Comparison of blaOXA-114 PCR Amplification, Multilocus Sequence Typing, and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

    Science.gov (United States)

    Rodrigues, Elenice R. A.; Ferreira, Alex G.; Leão, Robson S.; Leite, Cassiana C. F.; Carvalho-Assef, Ana Paula; Albano, Rodolpho M.

    2015-01-01

    Molecular methodologies were used to identify 28 Achromobacter spp. from patients with cystic fibrosis (CF). Multilocus sequence typing (MLST) identified 17 Achromobacter xylosoxidans isolates (all blaOXA-114 positive), nine Achromobacter ruhlandii isolates (all blaOXA-114 positive), one Achromobacter dolens isolate, and one Achromobacter insuavis isolate. All less common species were misidentified as A. xylosoxidans by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS). Chronic colonization by clonally related A. ruhlandii isolates was demonstrated. PMID:26400790

  4. MoS2/Ag nanohybrid: A novel matrix with synergistic effect for small molecule drugs analysis by negative-ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Zhao, Yaju; Deng, Guoqing; Liu, Xiaohui; Sun, Liang; Li, Hui; Cheng, Quan; Xi, Kai; Xu, Danke

    2016-09-21

    This paper reports a facile synthesis of molybdenum disulfide nanosheets/silver nanoparticles (MoS2/Ag) hybrid and its use as an effective matrix in negative ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The nanohybrid exerts a strong synergistic effect, leading to high performance detection of small molecule analytes including amino acids, peptides, fatty acids and drugs. The enhancement of laser desorption/ionization (LDI) efficiency is largely attributed to the high surface roughness and large surface area for analyte adsorption, better dispersibility, increased thermal conductivity and enhanced UV energy absorption as compared to pure MoS2. Moreover, both Ag nanoparticles and the edge of the MoS2 layers function as deprotonation sites for proton capture, facilitating the charging process in negative ion mode and promoting formation of negative ions. As a result, the MoS2/Ag nanohybrid proves to be a highly attractive matrix in MALDI-TOF MS, with desired features such as high desorption/ionization efficiency, low fragmentation interference, high salt tolerance, and no sweet-spots for mass signal. These characteristic properties allowed for simultaneous analysis of eight different drugs and quantification of acetylsalicylic acid in the spiked human serum. This work demonstrates for the first time the fabrication and application of a novel MoS2/Ag hybrid, and provides a new platform for use in the rapid and high throughput analysis of small molecules by mass spectrometry.

  5. Analysis of metal-binding proteins separated by non-denaturating gel electrophoresis using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS).

    Science.gov (United States)

    Becker, J Susanne; Mounicou, Sandra; Zoriy, Miroslav V; Becker, J Sabine; Lobinski, Ryszard

    2008-09-15

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) have become established as very efficient and sensitive biopolymer and elemental mass spectrometric techniques for studying metal-binding proteins (metalloproteins) in life sciences. Protein complexes present in rat tissues (liver and kidney) were separated in their native state in the first dimension by blue native gel electrophoresis (BN-PAGE). Essential and toxic metals, such as zinc, copper, iron, nickel, chromium, cadmium and lead, were detected by scanning the gel bands using quadrupole LA-ICP-MS with and without collision cell as a microanalytical technique. Several proteins were identified by using MALDI-TOF-MS together with a database search. For example, on one protein band cut from the BN-PAGE gel and digested with the enzyme trypsin, two different proteins - protein FAM44B and cathepsin B precursor - were identified. By combining biomolecular and elemental mass spectrometry, it was possible to characterize and identify selected metal-binding rat liver and kidney tissue proteins.

  6. Rapid identification of bacillus anthracis spores in suspicious powder samples by using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)

    NARCIS (Netherlands)

    Dybwad, M.; Laaken, A.L. van der; Blatny, J.M.; Paauw, A.

    2013-01-01

    Rapid and reliable identification of Bacillus anthracis spores in suspicious powders is important to mitigate the safety risks and economic burdens associated with such incidents. The aim of this study was to develop and validate a rapid and reliable laboratory- based matrix-assisted laser desorptio

  7. Substrate-Enhanced Micro Laser Desorption Ionization Mass Spectrometry Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Aerodyne Research, Inc. and the University of Massachusetts at Amherst will collaborate to develop laser desorption ionization (LDI) mass spectrometric analysis of...

  8. The high-mass component (>m/z 10 000) of coal tar pitch by matrix-assisted laser desorption/ionisation mass spectrometry and size-exclusion chromatography.

    Science.gov (United States)

    Millan, Marcos; Morgan, Trevor J; Behrouzi, Mahtab; Karaca, Fatma; Galmes, Carolina; Herod, Alan A; Kandiyoti, Rafael

    2005-01-01

    The size-exclusion chromatography (SEC) of acetone-soluble, pyridine-soluble and pyridine-insoluble fractions of a coal tar pitch indicates a bimodal distribution in each fraction. The proportion of high-mass material excluded from the SEC column porosity increases with solvent polarity. The polymer calibration of SEC shows the mass range of the small molecules to be from approximately 100 u to approximately 6000 u, with the mass range of the large excluded molecules above 200 000 u and up to several million u. In contrast, matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS) shows a similar low-mass range of ion abundances (< m/z 6000), but with a smaller range of high-mass ion abundances, from approximately m/z 10 000 to 100 000. The large molecules may have three-dimensional structures to allow molecules of relatively low mass to behave as if they are of large size in SEC. Laser desorption mass spectrometry of the acetone- and pyridine-soluble fractions produced molecular ions of polycyclic aromatics that can be related to the known compositions from gas chromatography (GC) mass spectrometry. The experimental conditions used to generate the bimodal distribution by MALDI-MS involve reducing the ion signal intensities to avoid overload of the detector and enable detection of the high-mass ions, by reducing the high-mass detector voltage (i.e. sensitivity) and increasing the laser power.

  9. How Suitable is Matrix-Assisted Laser Desorption/Ionization-Time-of-Flight for Metabolite Imaging from Clinical Formalin-Fixed and Paraffin-Embedded Tissue Samples in Comparison to Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance Mass Spectrometry?

    Science.gov (United States)

    Buck, Achim; Balluff, Benjamin; Voss, Andreas; Langer, Rupert; Zitzelsberger, Horst; Aichler, Michaela; Walch, Axel

    2016-05-17

    In research and clinical settings, formalin-fixed and paraffin-embedded (FFPE) tissue specimens are collected routinely and therefore this material constitutes a highly valuable source to gather insight in metabolic changes of diseases. Among mass spectrometry techniques to examine the molecular content of FFPE tissue, mass spectrometry imaging (MSI) is the most appropriate when morphological and histological features are to be related to metabolic information. Currently, high-resolution mass spectrometers are widely used for metabolomics studies. However, with regards to matrix-assisted laser desorption/ionization (MALDI) MSI, no study has so far addressed the necessity of instrumental mass resolving power in terms of clinical diagnosis and prognosis using archived FFPE tissue. For this matter we performed for the first time a comprehensive comparison between a high mass resolution Fourier-transform ion cyclotron resonance (FTICR) mass spectrometer and a time-of-flight (TOF) instrument with lower mass resolving power. Spectra analysis revealed that about one-third of the detected peaks remained unresolved by MALDI-TOF, which led to a 3-5 times lower number of m/z features compared to FTICR measurements. Overlaid peak information and background noise in TOF images made a precise assignment of molecular attributes to morphological features more difficult and limited classification approaches. This clearly demonstrates the need for high-mass resolution capabilities for metabolite imaging. Nevertheless, MALDI-TOF allowed reproducing and verifying individual markers identified previously by MALDI-FTICR MSI. The systematic comparison gives rise to a synergistic combination of the different MSI platforms for high-throughput discovery and validation of biomarkers.

  10. The laser desorption/laser ionization mass spectra of some methylated xanthines and the laser desorption of caffeine and theophylline from thin layer chromatography plates

    Science.gov (United States)

    Rogers, Kevin; Milnes, John; Gormally, John

    1993-02-01

    Laser desorption/laser ionization time-of-flight mass spectra of caffeine, theophylline, theobromine and xanthine are reported. These mass spectra are compared with published spectra obtained using electron impact ionization. Mass spectra of caffeine and theophylline obtained by IR laser desorption from thin layer chromatography plates are also described. The laser desorption of materials from thin layer chromatography plates is discussed.

  11. Modified-Atmospheric Pressure-Matrix Assisted Laser Desorption/Ionization Identification of Friction Modifier Additives Oleamide and Ethoxylated Tallow Amines on Varied Metal Target Materials and Tribologically Stressed Steel Surfaces.

    Science.gov (United States)

    Widder, Lukas; Ristic, Andjelka; Brenner, Florian; Brenner, Josef; Hutter, Herbert

    2015-11-17

    For many tasks in failure and damage analysis of surfaces deteriorated in heavy tribological contact, the detailed characterization of used lubricants and their additives is essential. The objective of the presented work is to establish accessibility of tribostressed surfaces for direct characterization via modified atmospheric pressure-matrix assisted laser desorption/ionization-mass spectrometry (m-AP-MALDI-MS). Special target holders were constructed to allow target samples of differing shape and form to fit into the desorption/ionization chamber. The best results of desorption and ionization on different target materials and varying roughnesses were achieved on smooth surfaces with low matrix/substrate interaction. M-AP-MALDI characterization of tribologically stressed steel surfaces after pin-on-disc sliding wear tests (SRV-tribotests) yielded positive identification of used friction modifier additives. Further structure elucidation by electrospray ionization mass spectrometry (ESI-MS) and measurements of worn surfaces by time-of-flight-secondary ion mass spectrometry (TOF-SIMS) accompanied findings about additive behavior and deterioration during tribological contact. Using m-AP-MALDI for direct offline examinations of worn surfaces may set up a quick method for determination of additives used for lubrication and general characterization of a tribological system.

  12. Studies on extracting solutions of endohedral rare-earth metallofullerenes by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    孙大勇; 刘志强; 刘子阳; 郭兴华; 徐文国; 刘淑莹

    1997-01-01

    Thirteen extracting solutions of rare-earth metallofullerenes containing La,Ce,Pr,Nd Sm,Eu,Gd,Tb,Dy,Ho,Er,Tm and Yb respectively have been investigated by means of matrix-assisted laser desorpuon/ ionization time-of-flight mass spectrometry.The influences of the positive-ion/negative-ion mode,laser intensity,ma trix and mass discrimination to the analytical results are studied,based on which the optimal analytical conditions have been determined.The results show that the extracting solutions contain large quantities of rare-earth metallofullerenes besides empty fullerenes.On the basis of comparing their relative intensities,the different structure stabilities and solubilities of metallofullerenes with different rare-earth metals encapsulated into the fullerene cages,as well as some possible reasons to those differences,are discussed.

  13. Impact of rapid identification of Acinetobacter Baumannii via matrix-assisted laser desorption ionization time-of-flight mass spectrometry combined with antimicrobial stewardship in patients with pneumonia and/or bacteremia.

    Science.gov (United States)

    Wenzler, Eric; Goff, Debra A; Mangino, Julie E; Reed, Erica E; Wehr, Allison; Bauer, Karri A

    2016-01-01

    We evaluated the clinical and economic outcomes of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) with stewardship intervention in patients with Acinetobacter baumannii (AB) pneumonia and/or bacteremia. 66 patients were included in the pre-intervention group and 53 in the intervention group. The combination of AB identification via MALDI-TOF MS and ID PharmD intervention significantly reduced the median time to effective therapy compared to conventional identification without intervention [77.7 (95% CI: 73.1-84.8) to 36.6 (95% CI: 25.9-50.9) hours (P timely, effective antimicrobial therapy and is associated with increased clinical cure.

  14. Development of a Rapid and Accurate Identification Method for Citrobacter Species Isolated from Pork Products Using a Matrix-Assisted Laser-Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS).

    Science.gov (United States)

    Kwak, Hye-Lim; Han, Sun-Kyung; Park, Sunghoon; Park, Si Hong; Shim, Jae-Yong; Oh, Mihwa; Ricke, Steven C; Kim, Hae-Yeong

    2015-09-01

    Previous detection methods for Citrobacter are considered time consuming and laborious. In this study, we have developed a rapid and accurate detection method for Citrobacter species in pork products, using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). A total of 35 Citrobacter strains were isolated from 30 pork products and identified by both MALDI-TOF MS and 16S rRNA gene sequencing approaches. All isolates were identified to the species level by the MALDI-TOF MS, while 16S rRNA gene sequencing results could not discriminate them clearly. These results confirmed that MALDI-TOF MS is a more accurate and rapid detection method for the identification of Citrobacter species.

  15. Determination of self-exchange rate of alkanethiolates in self-assembled monolayers on gold using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Kang, Hyunook; Kim, Yongbin; Choi, Inseong; Chang, Rakwoo; Yeo, Woon-Seok

    2014-09-16

    In this paper, we describe a new method for determining the exchange rates of alkanethiolates in self-assembled monolayers (SAMs) on gold using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to analyze the compositions of the alkanethiolate in SAMs rapidly and directly. In particular, to investigate the self-exchange of alkanethiols, we prepared a deuterated alkanethiol that has the same molecular properties as the non-deuterated alkanethiol but a different molecular weight. SAMs consisting of deuterated alkanethiolates were immersed in a solution of the non-deuterated alkanethiol, and the influences of the immersion time, temperature, concentration, and solvent on the self-exchange rates were investigated. Furthermore, we assessed the exchange rates among alkanethiols with different carbon chain lengths and different size of ethylene glycol units. In addition, we performed molecular dynamics simulations using a model SAM system in order to understand the molecular mechanism of the exchange process.

  16. A microwave-mediated saponification of galactosylceramide and galactosylceramide I3-sulfate and identification of their lyso-compounds by delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Taketomi, T; Hara, A; Uemura, K; Kurahashi, H; Sugiyama, E

    1996-07-16

    Small amounts of galactosylceramide (cerebroside) and galactosylceramide I3-sulfate (sulfatide) obtained from porcine spinal cord and equine kidney were deacylated by a rapid method of microwave-mediated saponification to prepare their lyso-compounds. Mass spectra of their protonated or deprotonated molecular ion peaks were detected by recently developed new technology of a delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometer with reflector detector in positive or negative ion mode. Long chain bases of lysocerebroside and lysosulfatide were different between porcine spinal cord and equine kidney, but similar to each other in the same organ, suggesting their common synthetic pathway. It is noted that the new rapid method can be similarly applied to the deacylation of both cerebroside and sulfatide in contrast to our classical method which was able to be applied to cerebroside, but not to sulfatide.

  17. Sensitive Detection of Aromatic Hydrophobic Compounds in Water and Perfluorooctane Sulfonate in Human Serum by Surface-Assisted Laser Desorption/Ionization Mass Spectrometry (SALDI-MS) with Amine Functionalized Graphene-Coated Cobalt Nanoparticles.

    Science.gov (United States)

    Nakai, Keisuke; Kawasaki, Hideya; Yamamoto, Atsushi; Arakawa, Ryuichi; Grass, Robert N; Stark, Wendelin J

    2014-01-01

    In this article, we describe the application of surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) with the use of amine functionalized graphene-coated cobalt nanoparticles (CoC-NH2 nanoparticles) to analyse aromatic hydrophobic compounds that are known environmental contaminants, including polycyclic aromatic hydrocarbons (PAHs) and pentachlorophenol (PCP). Our results demonstrated that SALDI-MS can detect PCP, anthracene, and pyrene in water. In particular, the CoC-NH2 nanoparticles proved to be an efficient means of capturing PCP in water because of the high adsorption capacity of the nanoparticles for PCP, which resulted in a detectability of 100 ppt. Furthermore, the CoC-NH2 nanoparticles also functioned as an adsorbent for solid-phase extraction of perfluorooctane sulfonate (PFOS) from human serum, displaying good performance with a detectability of 10 ppb by SALDI-MS.

  18. Recognition of Streptococcus pseudoporcinus Colonization in Women as a Consequence of Using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Group B Streptococcus Identification.

    Science.gov (United States)

    Suwantarat, Nuntra; Grundy, Maureen; Rubin, Mayer; Harris, Renee; Miller, Jo-Anne; Romagnoli, Mark; Hanlon, Ann; Tekle, Tsigereda; Ellis, Brandon C; Witter, Frank R; Carroll, Karen C

    2015-12-01

    During a 14-month period of using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for group B streptococcus (GBS) identification, we recovered 32 (1%) Streptococcus pseudoporcinus isolates from 3,276 GBS screening cultures from female genital sources (25 isolates from pregnant women and 7 from nonpregnant women). An additional two S. pseudoporcinus isolates were identified from a urine culture and a posthysterectomy wound culture. These isolates were found to cross-react with three different GBS antigen agglutination kits, PathoDx (Remel) (93%), Prolex (Pro-Lab Diagnostics) (38%), and Streptex (Remel) (53%). New approaches to bacterial identification in routine clinical microbiology laboratories may affect the prevalence of S. pseudoporcinus.

  19. Amazonian vegetable oils and fats: fast typification and quality control via triacylglycerol (TAG) profiles from dry matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry fingerprinting.

    Science.gov (United States)

    Saraiva, Sérgio A; Cabral, Elaine C; Eberlin, Marcos N; Catharino, Rodrigo R

    2009-05-27

    Amazonian oils and fats display unique triacylglycerol (TAG) profiles and, because of their economic importance as renewable raw materials and use by the cosmetic and food industries, are often subject to adulteration and forgery. Representative samples of these oils (andiroba, Brazil nut, buriti, and passion fruit) and fats (cupuaçu, murumuru, and ucuúba) were characterized without pre-separation or derivatization via dry (solvent-free) matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Characteristic profiles of TAG were obtained for each oil and fat. Dry MALDI-TOF MS provides typification and direct and detailed information, via TAG profiles, of their variable combinations of fatty acids. A database from spectra could be developed and may be used for their fast and reliable typification, application screening, and quality control.

  20. Rapid Profiling of Bovine and Human Milk Gangliosides by Matrix-Assisted Laser Desorption/Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry.

    Science.gov (United States)

    Lee, Hyeyoung; An, Hyun Joo; Lerno, Larry A; German, J Bruce; Lebrilla, Carlito B

    2011-08-15

    Gangliosides are anionic glycosphingolipids widely distributed in vertebrate tissues and fluids. Their structural and quantitative expression patterns depend on phylogeny and are distinct down to the species level. In milk, gangliosides are exclusively associated with the milk fat globule membrane. They may participate in diverse biological processes but more specifically to host-pathogen interactions. However, due to the molecular complexities, the analysis needs extensive sample preparation, chromatographic separation, and even chemical reaction, which makes the process very complex and time-consuming. Here, we describe a rapid profiling method for bovine and human milk gangliosides employing matrix-assisted desorption/ionization (MALDI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS). Prior to the analyses of biological samples, milk ganglioside standards GM3 and GD3 fractions were first analyzed in order to validate this method. High mass accuracy and high resolution obtained from MALDI FTICR MS allow for the confident assignment of chain length and degree of unsaturation of the ceramide. For the structural elucidation, tandem mass spectrometry (MS/MS), specifically as collision-induced dissociation (CID) and infrared multiphoton dissociation (IRMPD) were employed. Complex ganglioside mixtures from bovine and human milk were further analyzed with this method. The samples were prepared by two consecutive chloroform/methanol extraction and solid phase extraction. We observed a number of differences between bovine milk and human milk. The common gangliosides in bovine and human milk are NeuAc-NeuAc-Hex-Hex-Cer (GD3) and NeuAc-Hex-Hex-Cer (GM3); whereas, the ion intensities of ganglioside species are different between two milk samples. Kendrick mass defect plot yields grouping of ganglioside peaks according to their structural similarities. Gangliosides were further probed by tandem MS to confirm the compositional and structural assignments

  1. Laser-induced thermal desorption of aniline from silica surfaces

    Science.gov (United States)

    Voumard, Pierre; Zenobi, Renato

    1995-10-01

    A complete study on the energy partitioning upon laser-induced thermal desorption of aniline from silica surfaces was undertaken. The measurements include characterization of the aniline-quartz adsorption system using temperature-programmed desorption, the extrapolation of quasiequilibrium desorption temperatures to the regime of laser heating rates on the order of 109-1010 K/s by computational means, measurement of the kinetic energy distributions of desorbing aniline using a pump-probe method, and the determination of internal energies with resonance-enhanced multiphoton ionization spectroscopy. The measurements are compared to calculations of the surface temperature rise and the resulting desorption rates, based on a finite-difference mathematical description of pulsed laser heating. While the surface temperature of laser-heated silica reaches about 600-700 K at the time of desorption, the translational temperature of laser-desorbed aniline was measured to be Tkin=420±60 K, Tvib was 360±60 K, and Trot was 350±100 K. These results are discussed using different models for laser-induced thermal desorption from surfaces.

  2. Recent progress in application of carbon nanomaterials in laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Wang, Jing; Liu, Qian; Liang, Yong; Jiang, Guibin

    2016-04-01

    Carbon nanomaterials have attracted great interest over past decades owing to their unique physical properties, versatile functionalization chemistry, and biological compatibility. In this article, we review recent progress in application of carbon nanomaterials in laser desorption/ionization mass spectrometry (LDI MS). Various types of carbon nanomaterials, including fullerenes, carbon nanotubes, graphene, carbon nanodots, nanodiamond, nanofibers, nanohorns, and their derivative forms, are involved. The applications of these materials as new matrices or probes in matrix-assisted or surface-enhanced laser desorption/ionization mass spectrometry (MALDI or SELDI MS) are discussed. Finally, we summarize current challenges and give our perspectives on the future of applications of carbon nanomaterials in LDI MS.

  3. Facile Synthesis of N-Doped Carbon Dots as a New Matrix for Detection of Hydroxy-Polycyclic Aromatic Hydrocarbons by Negative-Ion Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry.

    Science.gov (United States)

    Lu, Wenjing; Li, Yong; Li, Ruijin; Shuang, Shaomin; Dong, Chuan; Cai, Zongwei

    2016-05-25

    N-doping carbon dots (N-CDs) were prepared by microwave-assisted pyrolysis of dl-malic acid and ethanolamine as precursors. The material served as an excellent matrix for the detection of the environmental pollutants hydroxy-polycyclic aromatic hydrocarbons (OH-PAHs) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in negative ion mode. The obtained N-CDs exhibited good UV absorption capacity and favorable solubility. The use of the N-CDs matrix exhibited low matrix background interference and was beneficial to improve the signal response due to the specific π-conjugated polyaromatic structure and the doping of nitrogen atoms. The developed method was found to have good reproducibility and sensitivity. The N-CDs as a new matrix also were employed for the detection of OH-PAHs in real PM2.5 samples. The mass concentrations of Σ-hydroxy-pyrene, Σ-dihydroxy-anthraquinone, and Σ-dihydroxy-benzo(a)pyrene on the collected PM2.5 samples ranged from 0.125 to 0.136 ng/m(3), 0.039 to 0.052 ng/m(3), and 0.053 to 0.072 ng/m(3), respectively. This work extends the application field of N-CDs and provides a good candidate of matrix for MALDI-TOF MS detection of environmental pollutants.

  4. Laser desorption of NO from a thick C 60 film

    Science.gov (United States)

    Hoger, T.; Marzok, C.; Jongma, R. T.; Zacharias, H.

    2006-09-01

    The desorption of NO molecules from a thick C 60 film is reported. A thermal desorption spectrum indicates two adsorption sites with binding energies of Eb = 0.30 eV and 0.55 eV. For laser desorption the fullerene surface is exposed to NO and excited by 7 ns UV laser pulses. Desorbing NO molecules are recorded state selectively as well as time resolved. The time-of-flight measurement indicates three different desorption pathways. A fast channel shows rovibronic temperatures of Trot( v″ = 0) = 370 K, Trot( v″ = 1) = 390 K and Tvib = 610 K as well as strong rotational-translational coupling. The desorption yield for the fast channel increases linearly with pulse energy with a desorption cross section of σ = (5.1 ± 0.9) × 10 -17 cm 2. Dominating the signal for small J″ values is a slow channel with low rotational and translational temperatures of about 110 K. We assign this peak to a laser-induced thermal desorption. For large pump-probe delays the data deviate from the Maxwellian flux distribution and a third channel appears with extremely late arrival times.

  5. Correlation of skin blanching and percutaneous absorption for glucocorticoid receptor agonists by matrix-assisted laser desorption ionization mass spectrometry imaging and liquid extraction surface analysis with nanoelectrospray ionization mass spectrometry.

    Science.gov (United States)

    Marshall, Peter; Toteu-Djomte, Valerie; Bareille, Philippe; Perry, Hayley; Brown, Gillian; Baumert, Mark; Biggadike, Keith

    2010-09-15

    Matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) and liquid extraction surface analysis (LESA) with nanoelectrospray ionization mass spectrometry (nESI-MS) have both been successfully employed to determine the degree of percutaneous absorption of three novel nonsteroid glucocorticoid receptor (GR) agonists in porcine ear sections. Historically, the ability of a glucocorticoid to elicit a skin blanching response when applied at low dose in ethanol solution to the forearms of healthy human volunteers has been a reliable predictor of their topical anti-inflammatory activity. While all three nonsteroidal GR agonists under investigation caused a skin blanching effect, the responses did not correlate with in vitro GR agonist potencies and different time courses were also observed for the skin blanching responses. MALDI MSI and LESA with nESI-MS were used to investigate and understand these different responses. The findings of the investigation was that the depth of porcine skin penetration correlates to the degree of skin blanching obtained for the same three compounds in human volunteers.

  6. Assessing the performance of novel software Strain Solution on automated discrimination of Escherichia coli serotypes and their mixtures using matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Ojima-Kato, Teruyo; Yamamoto, Naomi; Iijima, Yoshio; Tamura, Hiroto

    2015-12-01

    O157, O26, and O111 are the most important O serogroups of enterohemorrhagic Escherichia coli worldwide. Recently we reported a strategy for discriminating these serotypes from the others using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) based on the S10-spc-alpha operon gene-encoded ribosomal protein mass spectrum (S10-GERMS) method. To realize the fully automated identification of microorganisms at species- or serotype-level with the concept of S10-GERMS method, novel software named Strain Solution for MALDI-TOF MS was developed. In this study, the Strain Solution was evaluated with a total of 45 E. coli isolates including O26, O91, O103, O111, O115, O121, O128, O145, O157, O159, and untyped serotypes. The Strain Solution could accurately discriminate 92% (11/12) of O157 strains, 100% (13/13) of O26 and O111 strains from the others with three biomarkers in an automated manner. In addition, this software could identify 2 different E. coli strains (K-12 as a non-O157 representative and O157) in mixed samples. The results suggest that Strain Solution will be useful for species- or serotype-level classification of microorganisms in the fields of food safety and diagnostics.

  7. Metabolite localization by atmospheric pressure high-resolution scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging in whole-body sections and individual organs of the rove beetle Paederus riparius.

    Science.gov (United States)

    Bhandari, Dhaka Ram; Schott, Matthias; Römpp, Andreas; Vilcinskas, Andreas; Spengler, Bernhard

    2015-03-01

    Mass spectrometry imaging provides for non-targeted, label-free chemical imaging. In this study, atmospheric pressure high-resolution scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging (AP-SMALDI MSI) was used for the first time to describe the chemical distribution of the defensive compounds pederin, pseudopederin, and pederon in tissue sections (16 μm thick) of the rove beetle Paederus riparius. The whole-insect tissue section was scanned with a 20-μm step size. Mass resolution of the orbital trapping mass spectrometer was set to 100,000 at m/z 200. Additionally, organ-specific compounds were identified for brain, nerve cord, eggs, gut, ovaries, and malpighian tubules. To confirm the distribution of the specific compounds, individual organs from the insect were dissected, and MSI experiments were performed on the dissected organs. Three ganglia of the nerve cord, with a dimension of 250-500 μm, were measured with 10-μm spatial resolution. High-quality m/z images, based on high spatial resolution and high mass accuracy were generated. These features helped to assign mass spectral peaks with high confidence. Mass accuracy of the imaging experiments was pederin and its analogues could be visualized in the whole-insect section. Without any labeling, we assigned key lipids for specific organs to describe their location in the body and to identify morphological structures with a specificity higher than with staining or immunohistology methods.

  8. [Separation and identification of beta-casein from Chinese human milk by ion exchange chromatography-matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry].

    Science.gov (United States)

    Huang, Yu; Ren, Haowei; Liu, Biao; Liu, Ning; Li, Meng; Wang, Dongmao

    2013-05-01

    The selective precipitation of whole casein from skimmed milk was achieved by the addition of calcium salt under acidic pH. The effects of pH, centrifugal force and final concentration of CaCl2 on the separation of casein were studied by measuring the purity of final products using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that casein with the highest purity could be obtained with the pH of 4.3, the centrifugal force of 10 400 g and the final concentration of CaCl2 of 60 mmol/L. The casein was processed with DEAE anion exchange chromatography and three peaks were obtained. Then the third peak (peak III) was identified with Western-Blot method and matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry (MALDI-TOF/TOF MS). The identification of Western-Blot showed that peak III can combine with the specificity of human milk beta-casein antibody, and it is proved to be human milk beta-casein. The fingerprints of peak III were nalyzed by Mascot searching, and the sequence coverage was 50%, further supporting it is human milk beta-casein. In conclusion, an effective method to obtain human milk beta-casein from milk samples through DEAE anion exchange chromatography was established, and it is suitable for the proteomics research requirements of the beta-casein from human milk.

  9. The occurrence of Legionella species other than Legionella pneumophila in clinical and environmental samples in Denmark identified by mip gene sequencing and matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Svarrer, C W; Uldum, S A

    2012-10-01

    In Denmark, several laboratories use PCR as a routine diagnostic method for Legionnaires' disease, and almost all PCR-positive samples are investigated by culture. From 1993 to 2010, isolates of Legionella species other than Legionella pneumophila were obtained from respiratory samples from 33 patients, and from 1997 to 2010, 42 isolates of Legionella non-pneumophila species were obtained and saved from water samples from 39 different sites in Denmark. Macrophage infectivity potentiator gene (mip) sequencing was used as a reference method to identify the Legionella non-pneumophila species. Only one of the 75 isolates did not meet the acceptance criterion of a similarity of ≥98% to sequences in the database. The species distribution between clinical and environmental isolates varied. For the former, four species were detected, with Legionella bozemanae and Legionella micdadei predominating (both 44%). For the latter, eight species were detected, with Legionella anisa predominating (52%). The distribution among the Danish clinical isolates was different from the general distribution both in Europe and outside Europe, where L. bozemanae and Legionella longbeachae are the most commonly found clinical Legionella non-pneumophila species. The 75 isolates were also investigated by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS): 64 were correctly identified, with a score of ≥2.0; eight had a score of Legionella species identification.

  10. Comparison of an in-house method and the commercial Sepsityper™ kit for bacterial identification directly from positive blood culture broths by matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry.

    Science.gov (United States)

    Martiny, D; Dediste, A; Vandenberg, O

    2012-09-01

    The identification of bacteria directly from positive blood cultures using matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry (MALDI-TOF MS) is a new challenge to microbiologists. However, the protocols previously described are often difficult to implement in routine and comparisons are not always possible due to the variability of interpretative criteria. This study evaluated the analytical and practical performances of an in-house (IH) method, adapted from previous protocols, and the Sepsityper™ kit (Bruker Daltonics, Bremen, Germany). Positive blood cultures from 63 different patients were prospectively evaluated by both methods. To enhance the sensitivity of these methods, lowered cut-offs were assessed and validated on 66 additional samples. The IH method produced 86.4% and 73.7% correct genus and species identifications, respectively, when using the lowered cut-offs of 1.4 and 1.6 for correct genus and species identifications. The Sepsityper™ kit showed similar results (78.0% and 68.4% correct genus and species identification, respectively). However, the IH method is ten-fold less expensive than the commercial option (0.72 vs. 7.45 /analysis) and its turnaround time is approximately 20 min versus the nearly 40 min required for the Sepsityper™ kit, which includes an extraction step. Finally, the IH method was introduced twice-daily in our routine practice.

  11. Microorganisms in cryopreserved semen and culture media used in the in vitro production (IVP) of bovine embryos identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS).

    Science.gov (United States)

    Zampieri, Dávila; Santos, Vanessa G; Braga, Patrícia A C; Ferreira, Christina R; Ballottin, Daniela; Tasic, Ljubica; Basso, Andréa C; Sanches, Bruno V; Pontes, José H F; da Silva, Bárbara Pereira; Garboggini, Fabiana Fantinatti; Eberlin, Marcos N; Tata, Alessandra

    2013-09-01

    Commercial cattle breeders produce their own herd offspring for the dairy and beef market using artificial insemination. The procedure involves sanitary risks associated with the collection and commercialization of the germplasm, and the in vitro production and transfer of the bovine embryos must be monitored by strict health surveillance. To avoid the spreading of infectious diseases, one must rely on using controlled and monitored germplasm, media, and reagents that are guaranteed free of pathogens. In this article, we investigated the use of a new mass spectrometric approach for fast and accurate identification of bacteria and fungi in bovine semen and in culture media employed in the embryo in vitro production process. The microorganisms isolated from samples obtained in a commercial bovine embryo IVP setting were identified in a few minutes by their conserved peptide/protein profile, obtained applying matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), matched against a commercial database. The successful microorganisms MS identification has been confirmed by DNA amplification and sequencing. Therefore, the MS technique seems to offer a powerful tool for rapid and accurate microorganism identification in semen and culture media samples.

  12. Affinity-tagged phosphorylation assay by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (ATPA-MALDI): application to calcium/calmodulin-dependent protein kinase.

    Science.gov (United States)

    Kinumi, Tomoya; Niki, Etsuo; Shigeri, Yasushi; Matsumoto, Hiroyuki

    2005-12-01

    A matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based kinase assay using a peptide substrate tagged with a biotinyl group has been developed. The peptide moiety was designed to serve as an efficient substrate for calcium/calmodulin-dependent protein kinase II, based on the in vivo phosphorylation site of phosrestin I, a Drosophila homolog of arrestin. In the assay, the quantitative relationship was determined from the ratio of the peak areas between the two peaks respectively representing the unphosphorylated and the phosphorylated substrate. Attempts to assay phosphorylated peptides directly from the reaction mixture, gave inaccurate results because of the high noise level caused by the presence of salts and detergents. In contrast, after purifying the substrate peptides with the biotin affinity tag using streptavidin-coated magnetic beads, peak areas accurately represented the ratio between the unphosphorylated and phosphorylated peptide. By changing the substrate peptide to a peptide sequence that serves as a kinase substrate, it is expected that an efficient non-radioactive protein kinase assay using MALDI-TOF MS can be developed for any type of protein kinase. We call this technique "Affinity-Tagged Phosphorylation Assay by MALDI-TOF MS (ATPA-MALDI)." ATPA-MALDI should serve as a quick and efficient non-radioactive protein kinase assay by MALDI-TOF MS.

  13. Imipenem-avibactam: a novel combination for the rapid detection of carbapenemase activity in Enterobacteriaceae and Acinetobacter baumannii by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Oviaño, Marina; Bou, Germán

    2017-02-01

    In the present study, we propose a novel matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based method for detecting carbapenemase-producing Enterobacteriaceae and Acinetobacter baumannii. For this, we analyzed a series of 131 isolates. Among them, a total of 115 Enterobacteriaceae: 79 of them carrying a carbapenemase enzyme (15blaKPC, 7blaNDM, 11blaIMP, 12blaVIM, and 34blaOXA-48) and 16 A. baumannii isolates: 15 of them carrying carbapenemases (10blaOXA-23, 2blaOXA-58, 2blaOXA-24, and 1blaOXA-237). The rest of the isolates were noncarbapenemase producers and used as negative controls. The isolates were submitted to susceptibility testing using a combination of imipenem-avibactam and analysis by the MALDI-TOF Biotyper Compass software (Bruker Daltonik, Germany). The assay showed an overall sensitivity and specificity for carbapenemase detection of 98% and 100%, respectively. The combination of imipenem and avibactam displayed activity against KPC and OXA-48-producing Enterobacteriaceae and thus represents a new strategy for identifying and confirming these carbapenemases. However, the combination did not provide any benefit over A. baumannii.

  14. Graphene/TiO2 nanocomposite based solid-phase extraction and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for lipidomic profiling of avocado (Persea americana Mill.).

    Science.gov (United States)

    Shen, Qing; Yang, Mei; Li, Linqiu; Cheung, Hon-Yeung

    2014-12-10

    Phospholipids possess important physiological, structural and nutritional functions in biological systems. This study described a solid-phase extraction (SPE) method, employing graphene and titanium dioxide (G/TiO2) nanocomposite as sorbent, for the selective isolation and enrichment of phospholipids from avocado (Persea americana Mill.). Based on the principal that the phosphoryl group in the phospholipid can interact with TiO2 via a bridging bidentate mode, an optimum condition was established for SPE, and was successfully applied to prepare avocado samples. The extracts were monitored by matrix-assisted laser desorption ionization time-of-flight/tandem mass spectrometry (MALDI-TOF/MS) in both positive-ion and negative-ion modes. Results showed that phospholipids could be efficiently extracted in a clean manner by G/TiO2 based SPE. In addition, the signals of phospholipids were enhanced while the noise was reduced. Some minor peaks became more obvious. In conclusion, the nanocomposite material of G/TiO2 was proved to be a promising sorbent for selective separation of phospholipids from crude lipid extract.

  15. Evidence of genotypic diversity among Candida auris isolates by multilocus sequence typing, matrix-assisted laser desorption ionization time-of-flight mass spectrometry and amplified fragment length polymorphism.

    Science.gov (United States)

    Prakash, A; Sharma, C; Singh, A; Kumar Singh, P; Kumar, A; Hagen, F; Govender, N P; Colombo, A L; Meis, J F; Chowdhary, A

    2016-03-01

    Candida auris is a multidrug-resistant nosocomial bloodstream pathogen that has been reported from Asian countries and South Africa. Herein, we studied the population structure and genetic relatedness among 104 global C. auris isolates from India, South Africa and Brazil using multilocus sequence typing (MLST), amplified fragment length polymorphism (AFLP) fingerprinting and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). RPB1, RPB2 and internal transcribed spacer (ITS) and D1/D2 regions of the ribosomal DNA were sequenced for MLST. Further, genetic variation and proteomic assessment was carried out using AFLP and MALDI-TOF MS, respectively. Both MLST and AFLP typing clearly demarcated two major clusters comprising Indian and Brazilian isolates. However, the South African isolates were randomly distributed, suggesting different genotypes. MALDI-TOF MS spectral profiling also revealed evidence of geographical clustering but did not correlate fully with the genotyping methods. Notably, overall the population structure of C. auris showed evidence of geographical clustering by all the three techniques analysed. Antifungal susceptibility testing by the CLSI microbroth dilution method revealed that fluconazole had limited activity against 87% of isolates (MIC90, 64 mg/L). Also, MIC90 of AMB was 4 mg/L. Candida auris is emerging as an important yeast pathogen globally and requires reproducible laboratory methods for identification and typing. Evaluation of MALDI-TOF MS as a typing method for this yeast is warranted.

  16. Central nervous system disposition and metabolism of Fosdevirine (GSK2248761), a non-nucleoside reverse transcriptase inhibitor: an LC-MS and Matrix-assisted laser desorption/ionization imaging MS investigation into central nervous system toxicity.

    Science.gov (United States)

    Castellino, Stephen; Groseclose, M Reid; Sigafoos, James; Wagner, David; de Serres, Mark; Polli, Joseph W; Romach, Elizabeth; Myer, James; Hamilton, Brad

    2013-02-18

    The CNS disposition and metabolism of Fosdevirine (FDV), an HIV non-nucleoside reverse transcriptase inhibitor, was investigated in four patients who unexpectedly experienced seizures after at least 4 weeks of treatment in a Phase IIb, HIV-1 treatment experienced study. In addition, the CNS disposition and metabolism of FDV was examined in samples from rabbit, minipig, and monkey studies. LC-MS was used to characterize and estimate the concentrations of FDV and its metabolites in cerebral spinal fluid (seizure patients, rabbit, and monkey) and brain homogenate (rabbit, minipig, and monkey). The application of matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) provided the spatial distribution of FDV and its metabolites in brain tissue (rabbit, minipig, and monkey). A cysteine conjugate metabolite resulting from an initial glutathione (GSH) Michael addition to the trans-phenyl acrylonitrile moiety of FDV was the predominant drug-related component in the samples from seizure patients, rabbits, and minipigs. This metabolite persisted in the CNS for an extended period of time after the last dose in both seizure patients and minipigs. Furthermore, the localization of this metabolite was found to be highly associated with the white matter in rabbit and minipig brain sections by MALDI IMS. In contrast, the predominant component in monkey CNS was FDV, which was shown to be highly associated with the gray matter. On the basis of these data, several hypothesizes are considered, which might provide insights into species differences in CNS toxicity/seizures observed after FDV dosing.

  17. Analyses of black fungi by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS): species-level identification of clinical isolates of Exophiala dermatitidis.

    Science.gov (United States)

    Kondori, Nahid; Erhard, Marcel; Welinder-Olsson, Christina; Groenewald, Marizeth; Verkley, Gerard; Moore, Edward R B

    2015-01-01

    Conventional mycological identifications based on the recognition of morphological characteristics can be problematic. A relatively new methodology applicable for the identification of microorganisms is based on the exploitation of taxon- specific mass patterns recorded from abundant cell proteins directly from whole-cell preparations, using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). This study reports the application of MALDI-TOF MS for the differentiation and identifications of black yeasts, isolated from the respiratory tracts of patients with cystic fibrosis (CF). Initial phenotypic and DNA sequence-based analyses identified these isolates to be Exophiala dermatitidis. The type strains of E. dermatitidis (CBS 207.35(T)) and other species of Exophiala were included in the MALDI-TOF MS analyses to establish the references for comparing the mass spectra of the clinical isolates of Exophiala. MALDI-TOF MS analyses exhibited extremely close relationships among the clinical isolates and with the spectra generated from the type strain of E. dermatitidis. The relationships observed between the E. dermatitidis strains from the MALDI-TOF MS profiling analyses were supported by DNA sequence-based analyses of the rRNA ITS1 and ITS2 regions. These data demonstrated the applicability of MALDI-TOF MS as a reliable, rapid and cost-effective method for the identification of isolates of E. dermatitidis and other clinically relevant fungi and yeasts that typically are difficult to identify by conventional methods.

  18. Differentiation of vanA-positive Enterococcus faecium from vanA-negative E. faecium by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry.

    Science.gov (United States)

    Nakano, Satoshi; Matsumura, Yasufumi; Kato, Karin; Yunoki, Tomoyuki; Hotta, Go; Noguchi, Taro; Yamamoto, Masaki; Nagao, Miki; Ito, Yutaka; Takakura, Shunji; Ichiyama, Satoshi

    2014-09-01

    Vancomycin-resistant enterococci are important nosocomial pathogens that require rapid and accurate detection for infection control. Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS) has begun to be used in many clinical laboratories because it is a rapid, simple and inexpensive method for identifying micro-organisms. In this study, the performance of MALDI-TOF/MS to differentiate vanA-positive Enterococcus faecium (VPEF) from vanA-negative E. faecium (VNEF) was evaluated. A total of 61 VPEF isolates collected during regional surveillance in Kyoto (Japan) and 71 VNEF isolates collected from bacteraemia patients were analysed using MALDI-TOF/MS with three ClinProTools models. All of the isolates were correctly identified as E. faecium using the MALDI Biotyper system. To discriminate between VPEF and VNEF, all three ClinProTools models yielded >90% recognition capability (basic sensitivity) and cross-validation (reliability of the models); the genetic algorithm model exhibited the highest performance (99.18% and 92.40%, respectively). The high detection performance of MALDI-TOF/MS for VPEF offers the potential for routine laboratory use.

  19. Identification of blood culture isolates directly from positive blood cultures by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry and a commercial extraction system: analysis of performance, cost, and turnaround time.

    Science.gov (United States)

    Lagacé-Wiens, Philippe R S; Adam, Heather J; Karlowsky, James A; Nichol, Kimberly A; Pang, Paulette F; Guenther, Jodi; Webb, Amanda A; Miller, Crystal; Alfa, Michelle J

    2012-10-01

    Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry represents a revolution in the rapid identification of bacterial and fungal pathogens in the clinical microbiology laboratory. Recently, MALDI-TOF has been applied directly to positive blood culture bottles for the rapid identification of pathogens, leading to reductions in turnaround time and potentially beneficial patient impacts. The development of a commercially available extraction kit (Bruker Sepsityper) for use with the Bruker MALDI BioTyper has facilitated the processing required for identification of pathogens directly from positive from blood cultures. We report the results of an evaluation of the accuracy, cost, and turnaround time of this method for 61 positive monomicrobial and 2 polymicrobial cultures representing 26 species. The Bruker MALDI BioTyper with the Sepsityper gave a valid (score, >1.7) identification for 85.2% of positive blood cultures with no misidentifications. The mean reduction in turnaround time to identification was 34.3 h (P blood cultures and 26.5 h in a more practical setting where conventional identification or identification from subcultures was required for isolates that could not be directly identified by MALDI-TOF. Implementation of a MALDI-TOF-based identification system for direct identification of pathogens from blood cultures is expected to be associated with a marginal increase in operating costs for most laboratories. However, the use of MALDI-TOF for direct identification is accurate and should result in reduced turnaround time to identification.

  20. Direct identification of microorganisms from positive blood cultures using the lysis-filtration technique and matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS): a multicentre study.

    Science.gov (United States)

    Farina, Claudio; Arena, Fabio; Casprini, Patrizia; Cichero, Paola; Clementi, Massimo; Cosentino, Marina; Degl'Innocenti, Roberto; Giani, Tommaso; Luzzaro, Francesco; Mattei, Romano; Mauri, Carola; Nardone, Maria; Rossolini, Gian Maria; Serna Ortega, Paula Andrea; Vailati, Francesca

    2015-04-01

    Microbial identification from blood cultures is essential to institute optimal antibiotic therapy and improve survival possibilities. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been successfully applied to identify bacteria and yeasts from positive blood cultures broths. The aim of this multicentre study was to evaluate the reliability of the lysis-filtration technique associated with MALDI-TOF MS to directly identify microorganisms from 765 positive blood cultures collected in six Italian hospitals. Overall, 675/765 (78.1%) blood isolates were correctly identified at the species level, with significant differences between Gram-negative and Gram-positive bacteria (92.6%, and 69.8%, respectively). Some difficulties arise in identifying Streptococcus pneumoniae, Staphylococcus aureus, yeasts and anaerobes. The lysis-filtration protocol is a suitable procedure in terms of performance in identifying microorganisms, but it is quite expensive and technically time-consuming since the time of filtration is not regular for all the samples. The application of the MALDI-TOF MS technique to the direct microbial identification from positive blood cultures is a very promising approach, even if more experience must be gained to minimize errors and costs.

  1. Matrix-assisted laser desorption-ionization mass spectrometry peptide mass fingerprinting for proteome analysis: identification efficiency after on-blot or in-gel digestion with and without desalting procedures.

    Science.gov (United States)

    Lamer, S; Jungblut, P R

    2001-03-10

    In theory, peptide mass fingerprinting by matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS) has the potential to identify all of the proteins detected by silver staining on gels. In practice, if the genome of the organism investigated is completely sequenced, using current techniques, all proteins stained by Coomassie Brilliant Blue can be identified. This loss of identification sensitivity of ten to hundred-fold is caused by loss of peptides by surface contacts. Therefore, we performed digestion and transfer of peptides in the lower microl range and reduced the number of steps. The peptide mix obtained from in-gel or on-blot digestion was analyzed directly after digestion or after concentration on POROS R2 beads. Eight protein spots of a 2-DE gel from Mycobacterium bovis BCG were identified using these four preparation procedures for MALDI-MS. Overall, on-blot digestion was as effective as in-gel digestion. Whereas higher signal intensities resulted after concentration, hydrophilic peptides are better detected by direct measurement of the peptide mix without POROS R2 concentration.

  2. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric micro-analysis: the first non-immunological alternative attempt to quantify gluten gliadins in food samples.

    Science.gov (United States)

    Camafeita, E; Alfonso, P; Mothes, T; Méndez, E

    1997-09-01

    The first epitope-independent procedure for rapidly quantifying gluten gliadins in food by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI/TOF-MS) based on the direct observation of the characteristic gliadin mass pattern is presented. This pattern was identified in both processed and unprocessed gluten-containing food samples. The procedure allows the micro-quantification of gluten in food samples below levels toxic for coeliac patients, with a linear response in the 0.4-10 mg per 100 g range and a high detection sensitivity similar to that of enzyme-linked immunosorbent assay (ELISA) systems. Food samples simultaneously analyzed by MALDI/TOF-MS and a highly sensitive laboratory-made sandwich ELISA revealed a good correlation between the two techniques. In addition, MALDI/TOF-MS provides a rapid screening system to determine the presence of gliadins in food samples by directly monitoring the occurrence of the protonated gliadin mass pattern. The procedure also permits the study of the alteration of gliadins in food during the baking process, providing data on the heart effect by changes in protein mass signals.

  3. New strategy for the determination of gliadins in maize- or rice-based foods matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: fractionation of gliadins from maize or rice prolamins by acidic treatment.

    Science.gov (United States)

    Hernando, Alberto; Valdes, Israel; Méndez, Enrique

    2003-08-01

    A procedure for determining small quantities of gliadins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) in gluten-free foods containing relatively large amounts of prolamin proteins from maize or rice is described. We report for the first time that gliadins, the ethanol-soluble wheat prolamin fraction, can be quantitatively solubilized in 1.0 M acetic acid, while the corresponding ethanol-soluble maize or rice prolamin fraction remains insoluble in acetic acid. We describe a methodology for the detection of gliadins in maize and rice foods based on a two-step procedure of extraction (60% aqueous ethanol followed by 1 M acetic acid). Subsequent MALDI-TOFMS analysis of the resulting acidic extract from these gluten-free foods clearly confirms the presence of a typical mass pattern corresponding to gliadin components, ranging from 30 to 45 kDa. Depending on the percentages of maize or rice flours employed in the elaboration of these foods, the combined procedure enables levels of gliadins from 100 to 400 ppm to be detected. The efficiency of this combined procedure corroborates enzyme-linked immunosorbent assay data for a large number of maize/rice gluten-free foods by means of direct visualization of the characteristic gliadin mass pattern in maize or rice foods.

  4. Method development for compositional analysis of low molecular weight poly(vinyl acetate) by matrix-assisted/laser desorption-mass spectrometry and its application to analysis of chewing gum.

    Science.gov (United States)

    Tisdale, Evgenia; Wilkins, Charles

    2014-04-11

    The influence of the sample preparation parameters (the choice of the solvent and of the matrix:analyte ratio) was investigated and optimal conditions were established for MALDI mass spectrometry analysis of the pristine low molecular weight polyvinyl acetate (PVAc). It was demonstrated that comparison of polymer's and solvent's Hansen solubility parameters could be used as a guide when choosing the solvent for MALDI sample preparation. The highest intensity PVAc signals were obtained when ethyl acetate was used as a solvent along with the lowest matrix-analyte ratio (2,5-dihydroxybenzoic acid was used as a matrix in all experiments). The structure of the PVAc was established with high accuracy using the matrix-assisted laser desorption/ionization-Fourier transform mass spectrometry (MALDI-FTMS) analysis. It was demonstrated that PVAc undergoes unimolecular decomposition by losing acetic acid molecules from its backbone under the conditions of FTMS measurements. Number and weight average molecular weights as well as polydispersity indices were determined with both MALDI-TOF and MALDI-FTMS methods. The sample preparation protocol developed was applied to the analysis of a chewing gum and the molecular weight and structure of the polyvinyl acetate present in the sample were established. Thus, it was shown that optimized MALDI mass spectrometry could be used successfully for characterization of polyvinyl acetate in commercially available chewing gum.

  5. {sup 252}Cf plasma desorption and laser desorption mass spectrometry for the determination of molecular weight distribution of coal derivatives

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, B.R.; Bartle, K.D.; Ross, A.B.; Herod, A.A.; Kandiyoti, R.; Larsen, J.W. [University of Leeds, Leeds (United Kingdom). School of Chemistry

    1999-11-01

    A detailed knowledge of the molecular mass (MM) distribution in coal and its derived products is essential for a fundamental understanding of coal structure, and of the processes occurring during coal conversion. Fractionation using size exclusion chromatography (s.e.c.) using N-methyl-2-pyrrolidinone as the mobile phase has been applied to such materials and has provided improved MM distributions. Absolute calibration has been provided using matrix assisted laser desorption ionisation mass spectrometry (MAl.d.I.-m.s.). An alternative method of volatilising and ionising large molecules for mass spectrometry (m.s.) is {sup 252}Cf plasma desorption ({sup 252}Cf p.d.-m.s.). This involves the use of energetic fission fragments from the decay of {sup 252}Cf and produces mass spectra consisting predominantly of molecular ions from a range of polymers and biomolecules. This has been used by other workers to determine the molecular weight distribution of heavy distillation residues obtained from coal liquefaction processes either unfractionated or fractionated into broad fractions. Generally, a good agreement was obtained between values of MM determined by {sup 252}Cf p.d.-m.s. and s.e.c. A comparison is reported of MM distribution determined by {sup 252}Cf p.d.-m.s. and laser desorption mass spectrometry (l.d.-m.s.) for narrower fractions separated by s.e.c. from a coal tar pitch. 19 refs., 4 figs., 1 tab.

  6. Multidrug-Resistant Candida auris Misidentified as Candida haemulonii: Characterization by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry and DNA Sequencing and Its Antifungal Susceptibility Profile Variability by Vitek 2, CLSI Broth Microdilution, and Etest Method

    OpenAIRE

    2015-01-01

    Candida auris is a multidrug-resistant yeast that causes a wide spectrum of infections, especially in intensive care settings. We investigated C. auris prevalence among 102 clinical isolates previously identified as Candida haemulonii or Candida famata by the Vitek 2 system. Internal transcribed spacer region (ITS) sequencing confirmed 88.2% of the isolates as C. auris, and matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) easily separated all related...

  7. Graphene coated silica applied for high ionization matrix assisted laser desorption/ionization mass spectrometry: A novel approach for environmental and biomolecule analysis.

    Science.gov (United States)

    Nasser Abdelhamid, Hani; Wu, Bo-Sgum; Wu, Hui-Fen

    2014-08-01

    The integration of nanotechnology with mass spectrometry for sensitive and selective detection of molecules is a hot/important field of research. Synthesis of graphene (G) coated with mesoporous silica (SiO2, G@SiO2) for mass spectrometric application has been demonstrated. For the first time, we proposed the significant role of surfactant that used during the synthesis of mesorporous silicate (SiO2) in mass spectrometry. It was noticed that G could initiate SiO2 via surfactants which work as initiators for further ionization. The porosity of SiO2 trapped the analytes that was released and ionized with the surfactant fragments. Undoubtedly, strong background interferences were present in the case of organic matrix, which greatly obscured the detection of low molecular weight compounds. G@SiO2 nanocomposite affords several advantages, such as the ability to detect small molecules (SiO2 is not only due to the large surface area but also due to high desorption/ionization efficiency of inevitably surfactant (cetyltrimethylammonium chloride, CATB). Unlike the conventional MALDI-MS, the G@SiO2-MS is capable of generating multiply charged polysaccharides. The present method was validated to detect surfactants with low limits of detection.

  8. Analysis of Single Nucleotide Polymorphism in Human Paraoxonase 1 Gene(Q192R) with Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    @@ Introduction Single nucleotide polymorphisms (SNPs) are the most abundant DNA markers in the human genome occurring at a frequency of one in every 500-1000 nucleotides[1]. A variety of methods have been used for the analysis of single nucleotide polymorphisms, including restriction fragment length polymorphism (RFLP) [2], direct sequencing by using laser-induced fluorescence detection[3], fluorescence energy transfer[4], MALDI-TOF MS combined with primer extension or invasive cleavage[5,6] , and fluorescence polarization[7].

  9. Laser Desorption Mass Spectrometry. II. Applications to Structural Analysis.

    Science.gov (United States)

    1982-02-02

    was unexpected, was the quaternary amine Safranin -O. Ions corresponding to (M-HC)- at m/z 313 dominate the negative ion mass spectrum. Two classes of...and cluster species were observed. The positive ion LD mass spectrum of safranin -O, shown in Figure 6, is tvnical of LD results for salts. The...Figure 6. Laser Desorption Positive-ion Spectrum of Safranin 0. Figure 7. Laser Desorntion Mass Spectra of Doubly Charged Organic Salts Top - N,N

  10. Laser desorption lamp ionization source for ion trap mass spectrometry.

    Science.gov (United States)

    Wu, Qinghao; Zare, Richard N

    2015-01-01

    A two-step laser desorption lamp ionization source coupled to an ion trap mass spectrometer (LDLI-ITMS) has been constructed and characterized. The pulsed infrared (IR) output of an Nd:YAG laser (1064 nm) is directed to a target inside a chamber evacuated to ~15 Pa causing desorption of molecules from the target's surface. The desorbed molecules are ionized by a vacuum ultraviolet (VUV) lamp (filled with xenon, major wavelength at 148 nm). The resulting ions are stored and detected in a three-dimensional quadrupole ion trap modified from a Finnigan Mat LCQ mass spectrometer operated at a pressure of ≥ 0.004 Pa. The limit of detection for desorbed coronene molecules is 1.5 pmol, which is about two orders of magnitude more sensitive than laser desorption laser ionization mass spectrometry using a fluorine excimer laser (157 nm) as the ionization source. The mass spectrum of four standard aromatic compounds (pyrene, coronene, rubrene and 1,4,8,11,15,18,22,25-octabutoxy-29H,31H-phthalocyanine (OPC)) shows that parent ions dominate. By increasing the infrared laser power, this instrument is capable of detecting inorganic compounds.

  11. Laser assisted electron dynamics

    CERN Document Server

    Bray, Alexander William

    2016-01-01

    We apply the convergent close-coupling (CCC) formalism to analyse the processes of laser assisted electron impact ionisation of He, and the attosecond time delay in the photodetachment of the H^{-} ion and the photoionisation of He. Such time dependent atomic collision processes are of considerable interest as experimental measurements on the relevant timescale (attoseconds 10^{-18} s) are now possible utilising ultrafast and intense laser pulses. These processes in particular are furthermore of interest as they are strongly influenced by many-electron correlations. In such cases their theoretical description requires a more comprehensive treatment than that offered by first order perturbation theory. We apply such a treatment through the use of the CCC formalism which involves the complete numeric solution of the integral Lippmann-Schwinger equations pertaining to a particular scattering event. For laser assisted electron impact ionisation of He such a treatment is of a considerably greater accuracy than the...

  12. Source-identifying biomarker ions between environmental and clinical Burkholderia pseudomallei using whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS.

    Directory of Open Access Journals (Sweden)

    Suthamat Niyompanich

    Full Text Available Burkholderia pseudomallei is the causative agent of melioidosis, which is an endemic disease in Northeast Thailand and Northern Australia. Environmental reservoirs, including wet soils and muddy water, serve as the major sources for contributing bacterial infection to both humans and animals. The whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (whole-cell MALDI-TOF MS has recently been applied as a rapid, accurate, and high-throughput tool for clinical diagnosis and microbiological research. In this present study, we employed a whole-cell MALDI-TOF MS approach for assessing its potency in clustering a total of 11 different B. pseudomallei isolates (consisting of 5 environmental and 6 clinical isolates with respect to their origins and to further investigate the source-identifying biomarker ions belonging to each bacterial group. The cluster analysis demonstrated that six out of eleven isolates were grouped correctly to their sources. Our results revealed a total of ten source-identifying biomarker ions, which exhibited statistically significant differences in peak intensity between average environmental and clinical mass spectra using ClinProTools software. Six out of ten mass ions were assigned as environmental-identifying biomarker ions (EIBIs, including, m/z 4,056, 4,214, 5,814, 7,545, 7,895, and 8,112, whereas the remaining four mass ions were defined as clinical-identifying biomarker ions (CIBIs consisting of m/z 3,658, 6,322, 7,035, and 7,984. Hence, our findings represented, for the first time, the source-specific biomarkers of environmental and clinical B. pseudomallei.

  13. Mass spectrometry based lipid(ome) analyzer and molecular platform: a new software to interpret and analyze electrospray and/or matrix-assisted laser desorption/ionization mass spectrometric data of lipids: a case study from Mycobacterium tuberculosis.

    Science.gov (United States)

    Sabareesh, Varatharajan; Singh, Gurpreet

    2013-04-01

    Mass Spectrometry based Lipid(ome) Analyzer and Molecular Platform (MS-LAMP) is a new software capable of aiding in interpreting electrospray ionization (ESI) and/or matrix-assisted laser desorption/ionization (MALDI) mass spectrometric data of lipids. The graphical user interface (GUI) of this standalone programme is built using Perl::Tk. Two databases have been developed and constituted within MS-LAMP, on the basis of Mycobacterium tuberculosis (M. tb) lipid database (www.mrl.colostate.edu) and that of Lipid Metabolites and Pathways Strategy Consortium (LIPID MAPS; www.lipidmaps.org). Different types of queries entered through GUI would interrogate with a chosen database. The queries can be molecular mass(es) or mass-to-charge (m/z) value(s) and molecular formula. LIPID MAPS identifier also can be used to search but not for M. tb lipids. Multiple choices have been provided to select diverse ion types and lipids. Satisfying to input parameters, a glimpse of various lipid categories and their population distribution can be viewed in the output. Additionally, molecular structures of lipids in the output can be seen using ChemSketch (www.acdlabs.com), which has been linked to the programme. Furthermore, a version of MS-LAMP for use in Linux operating system is separately available, wherein PyMOL can be used to view molecular structures that result as output from General Lipidome MS-LAMP. The utility of this software is demonstrated using ESI mass spectrometric data of lipid extracts of M. tb grown under two different pH (5.5 and 7.0) conditions.

  14. Yersinia enterocolitica in diagnostic fecal samples from European dogs and cats: identification by fourier transform infrared spectroscopy and matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Stamm, Ivonne; Hailer, Mandy; Depner, Barbara; Kopp, Peter A; Rau, Jörg

    2013-03-01

    Yersinia enterocolitica is the main cause of yersiniosis in Europe, one of the five main bacterial gastrointestinal diseases of humans. Beside pigs, companion animals, especially dogs and cats, were repeatedly discussed in the past as a possible source of pathogenic Y. enterocolitica. To investigate the presence and types of Y. enterocolitica in companion animals, a total of 4,325 diagnostic fecal samples from dogs and 2,624 samples from cats were tested. The isolates obtained were differentiated by using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and Fourier transform infrared spectroscopy (FT-IR). Isolated Y. enterocolitica strains were bioserotyped. The detection of the ail gene by PCR and confirmation by FT-IR were used as a pathogenicity marker. Y. enterocolitica strains were isolated from 198 (4.6%) of the dog and 8 (0.3%) of the cat fecal samples investigated. One hundred seventy-nine isolates from dogs were analyzed in detail. The virulence factor Ail was detected in 91.6% of isolates. Isolates of biotype 4 (54.7%) and, to a lesser extent, biotypes 2 (23.5%), 3 (11.2%), and 5 (2.2%) were detected. The remaining 8.4% of strains belonged to the ail-negative biotype 1A. All 7 isolates from cats that were investigated in detail were ail positive. These results indicate that companion animals could be a relevant reservoir for a broad range of presumptively human-pathogenic Y. enterocolitica types. MALDI-TOF MS and FT-IR proved to be valuable methods for the rapid identification of Y. enterocolitica, especially in regard to the large number of samples that were investigated in a short time frame.

  15. Species-Level Identification of Actinomyces Isolates Causing Invasive Infections: Multiyear Comparison of Vitek MS (Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry) to Partial Sequencing of the 16S rRNA Gene.

    Science.gov (United States)

    Lynch, T; Gregson, D; Church, D L

    2016-03-01

    Actinomyces species are uncommon but important causes of invasive infections. The ability of our regional clinical microbiology laboratory to report species-level identification of Actinomyces relied on molecular identification by partial sequencing of the 16S ribosomal gene prior to the implementation of the Vitek MS (matrix-assisted laser desorption ionization-time of flight mass spectrometry [MALDI-TOF MS]) system. We compared the use of the Vitek MS to that of 16S rRNA gene sequencing for reliable species-level identification of invasive infections caused by Actinomyces spp. because limited data had been published for this important genera. A total of 115 cases of Actinomyces spp., either alone or as part of a polymicrobial infection, were diagnosed between 2011 and 2014. Actinomyces spp. were considered the principal pathogen in bloodstream infections (n = 17, 15%), in skin and soft tissue abscesses (n = 25, 22%), and in pulmonary (n = 26, 23%), bone (n = 27, 23%), intraabdominal (n = 16, 14%), and central nervous system (n = 4, 3%) infections. Compared to sequencing and identification from the SmartGene Integrated Database Network System (IDNS), Vitek MS identified 47/115 (41%) isolates to the correct species and 10 (9%) isolates to the correct genus. However, the Vitek MS was unable to provide identification for 43 (37%) isolates while 15 (13%) had discordant results. Phylogenetic analyses of the 16S rRNA sequences demonstrate high diversity in recovered Actinomyces spp. and provide additional information to compare/confirm discordant identifications between MALDI-TOF and 16S rRNA gene sequences. This study highlights the diversity of clinically relevant Actinomyces spp. and provides an important typing comparison. Based on our analysis, 16S rRNA gene sequencing should be used to rapidly identify Actinomyces spp. until MALDI-TOF databases are optimized.

  16. Rapid identification of bacteria and yeasts from positive-blood-culture bottles by using a lysis-filtration method and matrix-assisted laser desorption ionization-time of flight mass spectrum analysis with the SARAMIS database.

    Science.gov (United States)

    Fothergill, Amy; Kasinathan, Vyjayanti; Hyman, Jay; Walsh, John; Drake, Tim; Wang, Yun F Wayne

    2013-03-01

    Rapid identification of microorganisms causing bloodstream infections directly from a positive blood culture would decrease the time to directed antimicrobial therapy and greatly improve patient care. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is a fast and reliable method for identifying microorganisms from positive culture. This study evaluates the performance of a novel filtration-based method for processing positive-blood-culture broth for immediate identification of microorganisms by MALDI-TOF with a Vitek MS research-use-only system (VMS). BacT/Alert non-charcoal-based blood culture bottles that were flagged positive by the BacT/Alert 3D system were included. An aliquot of positive-blood-culture broth was incubated with lysis buffer for 2 to 4 min at room temperature, the resulting lysate was filtered through a membrane, and harvested microorganisms were identified by VMS. Of the 259 bottles included in the study, VMS identified the organisms in 189 (73%) cultures to the species level and 51 (19.7%) gave no identification (ID), while 6 (2.3%) gave identifications that were considered incorrect. Among 131 monomicrobic isolates from positive-blood-culture bottles with one spot having a score of 99.9%, the IDs for 131 (100%) were correct to the species level. In 202 bottles where VMS was able to generate an ID, the IDs for 189 (93.6%) were correct to the species level, whereas the IDs provided for 7 isolates (3.5%) were incorrect. In conclusion, this method does not require centrifugation and produces a clean spectrum for VMS analysis in less than 15 min. This study demonstrates the effectiveness of the new lysis-filtration method for identifying microorganisms directly from positive-blood-culture bottles in a clinical setting.

  17. Rapid discrimination of Haemophilus influenzae, H. parainfluenzae, and H. haemolyticus by fluorescence in situ hybridization (FISH and two matrix-assisted laser-desorption-ionization time-of-flight mass spectrometry (MALDI-TOF-MS platforms.

    Directory of Open Access Journals (Sweden)

    Hagen Frickmann

    Full Text Available BACKGROUND: Due to considerable differences in pathogenicity, Haemophilus influenzae, H. parainfluenzae and H. haemolyticus have to be reliably discriminated in routine diagnostics. Retrospective analyses suggest frequent misidentifications of commensal H. haemolyticus as H. influenzae. In a multi-center approach, we assessed the suitability of fluorescence in situ hybridization (FISH and matrix-assisted laser-desorption-ionization time-of-flight mass-spectrometry (MALDI-TOF-MS for the identification of H. influenzae, H. parainfluenzae and H. haemolyticus to species level. METHODOLOGY: A strain collection of 84 Haemophilus spp. comprising 50 H. influenzae, 25 H. parainfluenzae, 7 H. haemolyticus, and 2 H. parahaemolyticus including 77 clinical isolates was analyzed by FISH with newly designed DNA probes, and two different MALDI-TOF-MS systems (Bruker, Shimadzu with and without prior formic acid extraction. PRINCIPAL FINDINGS: Among the 84 Haemophilus strains analyzed, FISH led to 71 correct results (85%, 13 uninterpretable results (15%, and no misidentifications. Shimadzu MALDI-TOF-MS resulted in 59 correct identifications (70%, 19 uninterpretable results (23%, and 6 misidentifications (7%, using colony material applied directly. Bruker MALDI-TOF-MS with prior formic acid extraction led to 74 correct results (88%, 4 uninterpretable results (5% and 6 misidentifications (7%. The Bruker MALDI-TOF-MS misidentifications could be resolved by the addition of a suitable H. haemolyticus reference spectrum to the system's database. In conclusion, no analyzed diagnostic procedure was free of errors. Diagnostic results have to be interpreted carefully and alternative tests should be applied in case of ambiguous test results on isolates from seriously ill patients.

  18. Misidentification of Aspergillus nomius and Aspergillus tamarii as Aspergillus flavus: characterization by internal transcribed spacer, β-Tubulin, and calmodulin gene sequencing, metabolic fingerprinting, and matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Tam, Emily W T; Chen, Jonathan H K; Lau, Eunice C L; Ngan, Antonio H Y; Fung, Kitty S C; Lee, Kim-Chung; Lam, Ching-Wan; Yuen, Kwok-Yung; Lau, Susanna K P; Woo, Patrick C Y

    2014-04-01

    Aspergillus nomius and Aspergillus tamarii are Aspergillus species that phenotypically resemble Aspergillus flavus. In the last decade, a number of case reports have identified A. nomius and A. tamarii as causes of human infections. In this study, using an internal transcribed spacer, β-tubulin, and calmodulin gene sequencing, only 8 of 11 clinical isolates reported as A. flavus in our clinical microbiology laboratory by phenotypic methods were identified as A. flavus. The other three isolates were A. nomius (n = 2) or A. tamarii (n = 1). The results corresponded with those of metabolic fingerprinting, in which the A. flavus, A. nomius, and A. tamarii strains were separated into three clusters based on ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC MS) analysis. The first two patients with A. nomius infections had invasive aspergillosis and chronic cavitary and fibrosing pulmonary and pleural aspergillosis, respectively, whereas the third patient had A. tamarii colonization of the airway. Identification of the 11 clinical isolates and three reference strains by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) showed that only six of the nine strains of A. flavus were identified correctly. None of the strains of A. nomius and A. tamarii was correctly identified. β-Tubulin or the calmodulin gene should be the gene target of choice for identifying A. flavus, A. nomius, and A. tamarii. To improve the usefulness of MALDI-TOF MS, the number of strains for each species in MALDI-TOF MS databases should be expanded to cover intraspecies variability.

  19. Use of matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry for bacterial monitoring in routine analysis at a drinking water treatment plant.

    Science.gov (United States)

    Sala-Comorera, Laura; Vilaró, Carles; Galofré, Belén; Blanch, Anicet R; García-Aljaro, Cristina

    2016-10-01

    The study of bacterial communities throughout a drinking water treatment plant could provide a basic understanding of the effects of water processing that could then be used to improve the management of such plants. However, it is necessary to develop new analytical techniques that are sufficiently efficient, robust and fast for their effective and useful application in routine analysis. The aim of this study is therefore to assess the performance of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), as compared to the PhenePlate™ system, for routine analysis in a drinking water treatment plant. To this end we studied a total of 277 colonies isolated in different seasons and from different points throughout the water treatment process, including: raw water, sand filtration, ultrafiltration, reverse osmosis and chlorination. The colonies were analysed using MALDI-TOF MS by direct deposition of the cells on the plate. The colonies were also biochemically fingerprinted using the PhenePlate™ system, clustered according to their similarity and a representative strain was selected for 16S rRNA gene sequencing and API(®) gallery-based identification. The use of MALDI-TOF MS was reliable compared to the PhenePlate™ system and has the advantage of being faster and relatively cheap. Bacteria typing by MALDI-TOF MS is therefore a promising method to replace conventional routine phenotypic methods for the identification of bacteria in drinking water laboratories, thanks to its robustness. The major limiting factor for MALDI-TOF MS is the lack of a suitable mass spectra database; although each laboratory can develop its own library. This methodology will provide a tracking tool for companies to use in risk management and the detection of possible failures in both the water treatment processes and the distribution network, as well as offering characterization of the intrinsic microbial populations.

  20. Evaluation of the Bruker Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) System for the Identification of Clinically Important Dermatophyte Species.

    Science.gov (United States)

    Karabıçak, Nilgün; Karatuna, Onur; İlkit, Macit; Akyar, Işın

    2015-10-01

    Dermatophytes can invade the stratum corneum of the skin and other keratinized tissues and are responsible for a broad diversity of diseases of skin, nails and hair. Although the standard identification of dermatophytoses depends on macroscopic and microscopic characterization of the colonies grown on special media, there are a number of limitations owing to intraspecies morphological variability, atypical morphology or interspecies morphological similarity which entails improvement in the identification methods. Matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a novel method which proved to be effective for rapid and reliable identification of dermatophytes grown in cultures when compared to conventional methods. We evaluated the performance of Bruker MALDI-TOF MS System (Bruker Daltonics, Germany) for identification of clinically relevant dermatophytes. In order to increase the identification capacity of the system, we created supplemental spectral database entries using ten reference dermatophyte strains (ten species in two genera). The utility of the generated database was then challenged using a total of 126 dermatophytes (115 clinical isolates and 11 additional reference strains). The results were evaluated by both manufacturer-recommended and lowered cutoff scores. MALDI-TOF MS provided correct identification in 122 (96.8 %) and 113 (89.7 %) of the isolates with the lowered scores and using the supplemented database, respectively, versus 65 (51.6 %) and 17 (13.5 %) correct identifications obtained by the unmodified database and recommended scores at the genus and species levels, respectively. Our results support the potential utility of MALDI-TOF MS as a routine tool for accurate and reliable identification of dermatophytes.

  1. The Technical and Biological Reproducibility of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) Based Typing: Employment of Bioinformatics in a Multicenter Study

    Science.gov (United States)

    Oberle, Michael; Wohlwend, Nadia; Jonas, Daniel; Maurer, Florian P.; Jost, Geraldine; Tschudin-Sutter, Sarah; Vranckx, Katleen; Egli, Adrian

    2016-01-01

    Background The technical, biological, and inter-center reproducibility of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI TOF MS) typing data has not yet been explored. The aim of this study is to compare typing data from multiple centers employing bioinformatics using bacterial strains from two past outbreaks and non-related strains. Material/Methods Participants received twelve extended spectrum betalactamase-producing E. coli isolates and followed the same standard operating procedure (SOP) including a full-protein extraction protocol. All laboratories provided visually read spectra via flexAnalysis (Bruker, Germany). Raw data from each laboratory allowed calculating the technical and biological reproducibility between centers using BioNumerics (Applied Maths NV, Belgium). Results Technical and biological reproducibility ranged between 96.8–99.4% and 47.6–94.4%, respectively. The inter-center reproducibility showed a comparable clustering among identical isolates. Principal component analysis indicated a higher tendency to cluster within the same center. Therefore, we used a discriminant analysis, which completely separated the clusters. Next, we defined a reference center and performed a statistical analysis to identify specific peaks to identify the outbreak clusters. Finally, we used a classifier algorithm and a linear support vector machine on the determined peaks as classifier. A validation showed that within the set of the reference center, the identification of the cluster was 100% correct with a large contrast between the score with the correct cluster and the next best scoring cluster. Conclusions Based on the sufficient technical and biological reproducibility of MALDI-TOF MS based spectra, detection of specific clusters is possible from spectra obtained from different centers. However, we believe that a shared SOP and a bioinformatics approach are required to make the analysis robust and reliable. PMID:27798637

  2. Genotyping for Glycophorin GYP(B-A-B) Hybrid Genes Using a Single Nucleotide Polymorphism-Based Algorithm by Matrix-Assisted Laser Desorption/Ionisation, Time-of-Flight Mass Spectrometry.

    Science.gov (United States)

    Wei, Ling; Lopez, Genghis H; Ji, Yanli; Condon, Jennifer A; Irwin, Darryl L; Luo, Guangping; Hyland, Catherine A; Flower, Robert L

    2016-10-01

    The genetic basis for five GP(B-A-B) MNS system hybrid glycophorin blood group antigens results from rearrangement between the homologous GYPA and GYPB genes. Each hybrid glycophorin displays a characteristic profile of antigens. Currently, no commercial serological reagents are currently available to serologically type for these antigens. The aim of this study was to develop a single nucleotide polymorphism (SNP) mapping genotyping technique to allow characterisation of various GYP(B-A-B) hybrid alleles. Matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry (MS) assays were designed to genotype five GYP(B-A-B) hybrid alleles. Eight nucleotide positions were targeted and incorporated into the SNP mapping protocol. The allelic frequencies were calculated using peak areas. Sanger sequencing was performed to resolve a GYP*Hop 3' breakpoint. Observed allelic peak area ratios either coincided with the expected ratio or were skewed (above or below) from the expected ratio with switching occurring at and after the expected break point to generate characteristic mass spectral plots for each hybrid. Sequencing showed that the GYP*Hop crossover in the intron 3 region, for this example, was identical to that for GYP*Bun reference sequence. An analytical algorithm using MALDI-TOF MS genotyping platform defined GYPA inserts for five GYP(B-A-B) hybrids. The SNP mapping technique described here demonstrates proof of concept that this technology is viable for genotyping hybrid glycophorins, GYP(A-B-A), GYP(A-B) and GYP(B-A), and addresses the gap in current typing technologies.

  3. Network generation enhances interpretation of proteomics data sets by a combination of two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry.

    Science.gov (United States)

    Wang, Xijun; Zhang, Aihua; Sun, Hui; Wu, Gelin; Sun, Wenjun; Yan, Guangli

    2012-10-21

    Recent advances in proteomic technologies have enabled us to create detailed protein-protein interaction maps in diseases. As the size of the interaction dataset increases, powerful computational methods are required in order to effectively interpret network models from large scale interactome data. In this study, we carried out comparative proteomics to construct and identify the proteins networks associated with hepatic injury (HI) which are largely unknown, as a case study. All proteins expressed were separated and identified by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight-time-of-flight mass spectrometry (MALDI-TOF/TOF MS). Protein-interacting networks and pathways were mapped using STRING analysis program. We have performed for the first time a comprehensive profiling of changes in protein expression of HI rats, to uncover the networks altered by treated with CCl(4). Identification of fifteen spots (seven over-expressed and eight under-expressed) were established by MALDI-TOF/TOF MS. These proteins were subjected to functional pathway analysis using STRING software for better understanding of the biological context of the identified proteins. It suggested that modulation of multiple vital physiological pathways including DNA repair process, cell apoptosis, oxidation reduction, signal transduction, metabolic process, intracellular signaling cascade, regulation of biological processes, cell communication, regulation of cellular process, and molecular transport. In summary, the present study provides the first protein-interacting network maps and novel insights into the biological responses and potential pathways of HI. The generation of protein interaction networks clearly enhances the interpretation of proteomic data, particularly in respect of understanding molecular mechanisms of panel protein biomarkers.

  4. Matrix-assisted laser desorption ionization-time of flight and comparative genomic analysis of M-18 group a Streptococcus strains associated with an acute rheumatic fever outbreak in northeast Italy in 2012 and 2013.

    Science.gov (United States)

    Gaibani, Paolo; Scaltriti, Erika; Foschi, Claudio; Baggio, Enrico; Tamburini, Maria Vittoria; Creti, Roberta; Pascucci, Maria Grazia; Fagioni, Marco; Ambretti, Simone; Comandatore, Francesco; Pongolini, Stefano; Landini, Maria Paola

    2015-05-01

    Acute rheumatic fever (ARF) is a postsuppurative sequela caused by Streptococcus pyogenes infections affecting school-age children. We describe here the occurrence of an ARF outbreak that occurred in Bologna province, northeastern Italy, between November 2012 and May 2013. Molecular analysis revealed that ARF-related group A Streptococcus (GAS) strains belonged to the M-18 serotype, including subtypes emm18.29 and emm18.32. All M-18 GAS strains shared the same antigenic profile, including SpeA, SpeB, SpeC, SpeL, SpeM, and SmeZ. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) analysis revealed that M-18 GAS strains grouped separately from other serotypes, suggesting a different S. pyogenes lineage. Single nucleotide polymorphisms and phylogenetic analysis based on whole-genome sequencing showed that emm18.29 and emm18.32 GAS strains clustered in two distinct groups, highlighting genetic variations between these subtypes. Comparative analysis revealed a similar genome architecture between emm18.29 and emm18.32 strains that differed from noninvasive emm18.0 strains. The major sources of differences between M-18 genomes were attributable to the prophage elements. Prophage regions contained several virulence factors that could have contributed to the pathogenic potential of emm18.29 and emm18.32 strains. Notably, phage ΦSPBO.1 carried erythrogenic toxin A gene (speA1) in six ARF-related M-18 GAS strains but not in emm18.0 strains. In addition, a phage-encoded hyaluronidase gene (hylP.2) presented different variants among M-18 GAS strains by showing internal deletions located in the α-helical and TSβH regions. In conclusion, our study yielded insights into the genome structure of M-18 GAS strains responsible for the ARF outbreak in Italy, thus expanding our knowledge of this serotype.

  5. Quantitative mass barcode-like image of nicotine in single longitudinally sliced hair sections from long-term smokers by matrix-assisted laser desorption time-of-flight mass spectrometry imaging.

    Science.gov (United States)

    Nakanishi, Toyofumi; Nirasawa, Takashi; Takubo, Takayuki

    2014-01-01

    The matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometric technique (IMS) offered a new breakthrough perspective in the analysis of drug abuse in forensic science; however, it only produced barcode-like images, semi-quantitative analysis. In order to develop intermittent monitoring by this IMS for forensic and medical sciences, it is important to quantitatively measure the contents of longitudinally sliced hair sections. We developed quantitative imaging mass spectrometry (QIMS) of nicotine (NC) in longitudinally sliced hairs by MALDI-IMS with the selected reaction monitoring mode using a labeled NC ((13)C3-NC) standard for the serially chronological monitoring and traceability of NC intake in heavy smokers. The calibration curve of NC/(13)C3-NC was virtually a linear equation at ranges from 1 to 50 ng/mL, the slope was 0.020, and the intercept was almost 0.023 and the R(2) was 0.9965. The limit of quantitation of NC was calculated as 1.6 ng/mg hair (an average weight of the hair would be assumed 0.06 mg/cm) by QIMS. Moreover, NC concentrations in two separate heavy smokers (n = 3) were 8.5 ± 1.2 and 34.5 ± 2.8 ng/mg hair, respectively, and covariations were ∼10% using a single hair. Quantitative mass barcode-like image of sliced section of hair allowed for the quantitative assessment of NC concentrations in long-term smokers similar to drugs and medicines during drug histories.

  6. Development of C60-based labeling reagents for the determination of low-molecular-weight compounds by matrix assisted laser desorption ionization mass (I): Determination of amino acids in microliter biofluids.

    Science.gov (United States)

    Wu, Pin; Xiao, Hua-Ming; Ding, Jun; Deng, Qian-Yun; Zheng, Fang; Feng, Yu-Qi

    2017-04-01

    Quantification of low molecular weight compounds (<800 Da) using matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI MS) is challenging due to the matrix signal interference at low m/z region and poor reproducibility of MS responses. In this study, a C60 labeling-MALDI MS strategy was proposed for the fast, sensitive and reliable determination of amino acids (AAs) in biofluids. An N-hydroxysuccinimide functionalized C60 was synthesized as the labeling reagent and added as an 880 Da tag to AAs; a carboxyl acid containing C60 was employed as the internal standards to normalize MS variations. This solved the inherent problems of MALDI MS for small molecule analysis. The entire analytical procedure-which consisted of simple protein precipitation and 10 min of derivatization in a microwave prior to the MALDI MS analysis-could be accomplished within 20 min with high throughput and great sample matrix tolerance. AA quantification showed good linearity from 0.7 to 70.0 μM with correlation coefficients (R) larger than 0.9954. The limits of detection were 70.0-300.0 fmol. Good reproducibility and reliability of the method were demonstrated by intra-day and inter-day precision with relative standard deviations less than 13.8%, and the recovery in biofluid ranged from 80.4% to 106.8%. This approach could be used in 1 μL of urine, serum, plasma, whole blood, and cerebrospinal fluid. Most importantly, the C60 labeling strategy is a universal approach for MALDI MS analysis of various LMW compounds because functionalized C60 is now available on demand.

  7. Comparison of peptide nucleic acid fluorescence in situ hybridization assays with culture-based matrix-assisted laser desorption/ionization-time of flight mass spectrometry for the identification of bacteria and yeasts from blood cultures and cerebrospinal fluid cultures.

    Science.gov (United States)

    Calderaro, A; Martinelli, M; Motta, F; Larini, S; Arcangeletti, M C; Medici, M C; Chezzi, C; De Conto, F

    2014-08-01

    Peptide nucleic acid fluorescence in situ hybridization (PNA FISH) is a molecular diagnostic tool for the rapid detection of pathogens directly from liquid media. The aim of this study was to prospectively evaluate PNA FISH assays in comparison with culture-based matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) identification, as a reference method, for both blood and cerebrospinal fluid (CSF) cultures, during a 1-year investigation. On the basis of the Gram stain microscopy results, four different PNA FISH commercially available assays were used ('Staphylococcus aureus/CNS', 'Enterococcus faecalis/OE', 'GNR Traffic Light' and 'Yeasts Traffic Light' PNA FISH assays, AdvanDx). The four PNA FISH assays were applied to 956 positive blood cultures (921 for bacteria and 35 for yeasts) and 11 CSF cultures. Among the 921 blood samples positive for bacteria, PNA FISH gave concordant results with MALDI-TOF MS in 908/921 (98.64%) samples, showing an agreement of 99.4% in the case of monomicrobial infections. As regards yeasts, the PNA FISH assay showed a 100% agreement with the result obtained by MALDI-TOF MS. When PNA FISH assays were tested on the 11 CSF cultures, the results agreed with the reference method in all cases (100%). PNA FISH assays provided species identification at least one work-day before the MALDI-TOF MS culture-based identification. PNA FISH assays showed an excellent efficacy in the prompt identification of main pathogens, yielding a significant reduction in reporting time and leading to more appropriate patient management and therapy in cases of sepsis and severe infections.

  8. Identification and comparative proteomic study of quail and duck egg white protein using 2-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry analysis.

    Science.gov (United States)

    Hu, S; Qiu, N; Liu, Y; Zhao, H; Gao, D; Song, R; Ma, M

    2016-05-01

    A proteomic study of egg white proteins from 2 major poultry species, namely quail (Coturnix coturnix) and duck (Anas platyrhynchos), was performed with comparison to those of chicken (Gallus gallus) through 2-dimensional polyacrylamide gel electrophoresis (2-DE) analysis. By using matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF MS/MS), 29 protein spots representing 10 different kinds of proteins as well as 17 protein spots designating 9 proteins were successfully identified in quail and duck egg white, respectively. This report suggested a closer relationship between quail and chicken egg white proteome patterns, whereas the duck egg white protein distribution on the 2-DE map was more distinct. In duck egg white, some well-known major proteins, such as ovomucoid, clusterin, extracellular fatty acid-binding protein precursor (ex-FABP), and prostaglandin D2 synthase (PG D2 synthase), were not detected, while two major protein spots identified as "deleted in malignant brain tumors 1" protein (DMBT1) and vitellogenin-2 were found specific to duck in the corresponding range on the 2-DE gel map. These interspecies diversities may be associated with the egg white protein functions in cell defense or regulating/supporting the embryonic development to adapt to the inhabiting environment or reproduction demand during long-term evolution. The findings of this work will give insight into the advantages involved in the application on egg white proteins from various egg sources, which may present novel beneficial properties in the food industry or related to human health.

  9. Determination of osteocalcin in meat and bone meal of bovine and porcine origin using matrix-assisted laser desorption ionization/time-of-flight mass spectrometry and high-resolution hybrid mass spectrometry.

    Science.gov (United States)

    Balizs, Gabor; Weise, Christoph; Rozycki, Christel; Opialla, Tobias; Sawada, Stefanie; Zagon, Jutta; Lampen, Alfonso

    2011-05-05

    A method has been developed for determining the origin of meat and bone meal (MBM) by detecting species-specific osteocalcin (OC) using matrix-assisted laser desorption ionization/time-of-flight (MALDI/TOF) and high-resolution hybrid mass spectrometry (HR-Q/TOF MS). The analysis is based on the detection of typical species-specific OC and its tryptic peptide fragments which differ in mass due to differences in the amino-acid sequences between species. After dissolving the MBM samples in EDTA buffer, purification after ultrafiltration was performed using two methods: solid-phase extraction using Zip-Tip C(18) or size exclusion coupled with reverse-phase chromatography. Fractions containing partially purified intact OC were analyzed using LC-Q/TOF and MALDI/TOF mass spectrometry. Species-specific OC was detected at the typical protonated and doubly protonated molecular ions. Furthermore, typical porcine- and bovine-derived tryptic fragments from MBM were detected after enzymatic digestion. In order to determine the underlying amino-acid sequences and to confirm the assignment to OC-derived peptides, MS/MS analysis was carried out. In conclusion, we were able to detect OC in bovine and porcine MBM with high sensitivity and the MS-based method described here by which total OC mass and marker peptides of digested OC are recorded can be used as an alternative approach to detect genus-specific differences in MBM and can be applied as a confirmatory method to mainly immunological osteocalcin screening methods.

  10. Determination of osteocalcin in meat and bone meal of bovine and porcine origin using matrix-assisted laser desorption ionization/time-of-flight mass spectrometry and high-resolution hybrid mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Balizs, Gabor, E-mail: gabor.balizs@bfr.bund.de [Federal Institute for Risk Assessment, Thielallee 88-92, D-14195 Berlin (Germany); Weise, Christoph [Freie Universitaet Berlin, Institute for Chemistry and Biochemistry, Thielallee 63, D-14195 Berlin (Germany); Rozycki, Christel; Opialla, Tobias; Sawada, Stefanie; Zagon, Jutta; Lampen, Alfonso [Federal Institute for Risk Assessment, Thielallee 88-92, D-14195 Berlin (Germany)

    2011-05-05

    A method has been developed for determining the origin of meat and bone meal (MBM) by detecting species-specific osteocalcin (OC) using matrix-assisted laser desorption ionization/time-of-flight (MALDI/TOF) and high-resolution hybrid mass spectrometry (HR-Q/TOF MS). The analysis is based on the detection of typical species-specific OC and its tryptic peptide fragments which differ in mass due to differences in the amino-acid sequences between species. After dissolving the MBM samples in EDTA buffer, purification after ultrafiltration was performed using two methods: solid-phase extraction using Zip-Tip C{sub 18} or size exclusion coupled with reverse-phase chromatography. Fractions containing partially purified intact OC were analyzed using LC-Q/TOF and MALDI/TOF mass spectrometry. Species-specific OC was detected at the typical protonated and doubly protonated molecular ions. Furthermore, typical porcine- and bovine-derived tryptic fragments from MBM were detected after enzymatic digestion. In order to determine the underlying amino-acid sequences and to confirm the assignment to OC-derived peptides, MS/MS analysis was carried out. In conclusion, we were able to detect OC in bovine and porcine MBM with high sensitivity and the MS-based method described here by which total OC mass and marker peptides of digested OC are recorded can be used as an alternative approach to detect genus-specific differences in MBM and can be applied as a confirmatory method to mainly immunological osteocalcin screening methods.

  11. Laser assisted {alpha} decay

    Energy Technology Data Exchange (ETDEWEB)

    Castaneda Cortes, Hector Mauricio

    2012-02-01

    Excited or short-lived nuclei often decay by emitting alpha particles that are assumed to be preformed inside the nucleus and confined in the nuclear potential well. In this picture, {alpha} decay refers to the tunneling of the alpha particle through the potential barrier. In this thesis we investigate for the first time how strong laser fields can assist the tunneling of the alpha particle and thus influence the nuclear decay. Generally speaking, laser-assisted {alpha} decay can be described as laser-assisted tunneling of a quasistationary state, i.e, a slowly decaying state. Our theoretical treatment is developed starting from the complex trajectory formulation of the well-known strong-field approximation used to describe laser-induced ionization. We extend this formulation and develop a method to treat the decay of quasistationary states. The effect of both static and optical and X-ray monochromatic fields on the lifetimes and {alpha}-particle emission spectra are investigated for a number of {alpha}-emitting nuclei. We find that even at strong intensities, the laser-induced acceleration of the {alpha} decay is negligible, ranging from a relative modification in the decay rate of 10{sup -3} for static fields of electric field strengths of 10{sup 15} V/m, to 10{sup -8} for strong optical fields with intensities of 10{sup 22} W/cm{sup 2}, and to 10{sup -6} for strong X-ray fields with laser intensities around 10{sup 24} W/cm{sup 2}. However, the effect of the external field is visible in the spectrum of emitted alpha particles, leading in the case of optical fields even to rescattering phenomena for intensities approaching 6 x 10{sup 22} W/cm{sup 2}. The dynamics of the alpha particle in laser fields of intensities below the rescattering limit is investigated.

  12. A comparison of nano-electrospray gas-phase electrophoretic mobility macromolecular analysis and matrix-assisted laser desorption/ionization linear time-of-flight mass spectrometry for the characterization of the recombinant coagulation glycoprotein von Willebrand factor.

    Science.gov (United States)

    Kemptner, Jasmin; Marchetti-Deschmann, Martina; Müller, Roland; Ivens, Andreas; Turecek, Peter; Schwarz, Hans Peter; Allmaier, Günter

    2010-03-01

    Von Willebrand factor (VWF), an adhesive glycoprotein with an approximate molecular weight (MW) of the monomer of 260 kDa, circulates in human blood plasma as a series of multimers ranging in size up to 20.000 kDa; thus the determination of the accurate MW of the monomer is of great importance and due to its high MW quite challenging. In this study accurate MW determination of intact recombinant VWF monomer (rVWF) was performed with GEMMA (gas-phase electrophoretic mobility macromolecular analysis) and MALDI TOF MS (matrix-assisted laser desorption/ionization linear time-of-flight mass spectrometry). Three rVWF preparations with differing buffer systems and glycoprotein concentrations were analyzed. First investigations directed towards heterogeneity determination by means of capillary gel electrophoresis (CGE)-on-the-chip with a laser-induced fluorescence detector revealed two compounds (MW of 277 kDa (migration time 44.3 s) and 341 kDa (migration time 49.5 s)) present in each sample to varying extents, namely mature and pro-rVWF. MALDI MS analysis in the linear positive ion mode allowed the detection of mature rVWF with an exact MW of 256.1 kDa (+/-0.8%) and pro-rVWF with a MW of 349.8 kDa (+/-0.8%). Two samples containing pro-rVWF in very minor concentration resulted in GEMMA detection of the mature rVWF with a MW of 227.4 kDa (+/-2.5%), derived from the measured globular size of 10.9 nm. For one sample containing both rVWF species in almost equal concentrations no differentiation of the two species was possible with GEMMA. Due to its lower resolution only a peak representing a mixture of both species at 11.8 nm could be observed, yielding a MW of 298.8 kDa (+/-1.6%).

  13. Laser Desorption Ionization Mass Spectrometry Imaging of Drosophila Brain Using Matrix Sublimation versus Modification with Nanoparticles.

    Science.gov (United States)

    Phan, Nhu T N; Mohammadi, Amir Saeid; Dowlatshahi Pour, Masoumeh; Ewing, Andrew G

    2016-02-02

    Laser desorption ionization mass spectrometry (LDI-MS) is used to image brain lipids in the fruit fly, Drosophila, a common invertebrate model organism in biological and neurological studies. Three different sample preparation methods, including sublimation with two common organic matrixes for matrix-assisted laser desorption ionization (MALDI) and surface-assisted laser desorption ionization (SALDI) using gold nanoparticles, are examined for sample profiling and imaging the fly brain. Recrystallization with trifluoroacetic acid following matrix deposition in MALDI is shown to increase the incorporation of biomolecules with one matrix, resulting in more efficient ionization, but not for the other matrix. The key finding here is that the mass fragments observed for the fly brain slices with different surface modifications are significantly different. Thus, these approaches can be combined to provide complementary analysis of chemical composition, particularly for the small metabolites, diacylglycerides, phosphatidylcholines, and triacylglycerides, in the fly brain. Furthermore, imaging appears to be beneficial using modification with gold nanoparticles in place of matrix in this application showing its potential for cellular and subcellular imaging. The imaging protocol developed here with both MALDI and SALDI provides the best and most diverse lipid chemical images of the fly brain to date with LDI.

  14. Thermal desorption from surfaces with laser-induced defects

    Energy Technology Data Exchange (ETDEWEB)

    Szabelski, Pawel; Panczyk, Tomasz; Rudzinski, Wladyslaw

    2002-12-30

    Monte Carlo simulation method was used to mimic surface damage development caused by short laser pulses. The influence of pulsed laser irradiation on the creation of defect concentration was examined in the case of a model surface. In particular, the dependence of the intact surface area on a number of laser scans was studied and compared with the experimental results obtained for Rh(1 1 1) crystal face. Changes in the adsorptivoperties of the surface produced by laser irradiation are explained with the help of a simple geometric model connecting the laser intensity and the disordered area generated by a single laser shot. It was demonstrated that exponential decay of the Low Energy Electron Diffraction (LEED) signal with the number of laser scans, which is observed experimentally, may result directly from the overlapping of the laser spots created on the surface. This effect becomes enhanced when the laser intensity, hence the spot size, increases. The importance of laser-induced defects in the kinetics of catalytic/separation processes was examined in the case of temperature programmed desorption (TPD) spectra from surfaces subjected to a different number of laser shots. The spectra were simulated by employing the Monte Carlo method as well as by application of the absolute rate theory (ART) coupled with the mean field approximation. The results obtained with both methods were in a good agreement even when weak lateral interactions in the adsorbed phase were allowed.

  15. Investigation of individual aerosol particles matrix-assisted laser desorption/lonization%单粒子气溶胶的基质辅助激光解吸电离研究

    Institute of Scientific and Technical Information of China (English)

    提汝芳; 张子良; 王颖萍; 丁蕾; 郑海洋; 方黎

    2012-01-01

    单粒子气溶胶的基质辅助激光解吸/电离飞行时间质谱技术具有快速在线检测环境大气生物气溶胶的潜力.以α-环糊精为分析物,实验研究了吡啶羧酸、2,5-二羟基苯甲酸、芥子酸、阿魏酸四种常用基质材料的效果,考察基质与分析物的摩尔比对分析物单粒子激光质谱信号的影响.实验结果表明阿魏酸做基质时质谱命中率及分析物质谱信号峰检测效率最高,吡啶羧酸做基质时的分析物质谱信号峰激发率最高,信号峰强度最强.吡啶羧酸对分析物摩尔比为100∶ 1时,分析物质谱信号检测效率最高.%Matrix-assisted laser desorption/ionization time-of-flight mass spectroraetry for individual aerosol has potential to detect atmospheric bioaerosols near real-time rapidly. Ananlyte a-Cyclodextrin and four matrices containing Picolinic acid, 2,5-Dihydroxybenzoic acid, sinapic acid and ferulic acid were investigated. Effect of matrix to analyte molar ratio on individual analyte signal mass spectra was analyzed. Experiments illustrated that hit rate and detection efficiency of analyte's signal mass spectra were hightest when ferulic acid as the matrix. Excitation efficiency and relative intensity of the cationized analyte ion signal were optimal while Picolinic acid as the matrix. Detection efficienc of ananlyte signal mass spectra was optimal when matrix to analyte molar ratio was 100 : 1.

  16. Applicability of an in-House Saponin-Based Extraction Method in Bruker Biotyper Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry System for Identification of Bacterial and Fungal Species in Positively Flagged Blood Cultures

    Directory of Open Access Journals (Sweden)

    Jung-Yien Chien

    2016-09-01

    Full Text Available We used an in-house saponin-based extraction method to evaluate the performance of the Bruker Biotyper matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS system for the identification of bacteria and fungi in 405 positively flagged blood culture bottles. Results obtained from MALDI-TOF/MS were compared with those obtained using conventional phenotypic identification methods. Of the 405 positively flagged blood culture bottles, 365 showed monomicrobal growth and were correctly identified to the species (72.1% or genus (89.6% level using the Bruker Biotyper system. The remaining 40 positively flagged blood culture bottles showed polymicrobial growth. Of them, 82.5% (n=33 of the isolates were correctly identified to the species level and 92.5% (n=37 to the genus level using the Bruker Biotyper system. The overall accuracy of identification to the genus level in flagged blood cultures was 89.5% for Gram-positive organisms, 93.5% for Gram-negative pathogens and 71.9% for fungi. Confidence scores were 1.500 for 307 (75.8% bottles, 1.700 for 249 (61.5% bottles and 2.000 for 142 (35.1% bottles. None of the yeast cultures yielded scores 1.700. Using an identification-score cutoff of 1.500, the MALDI Biotyper correctly identified 99.2% of Gram-positive bacteria, 97.6% of Gram-negative bacteria and 100% of yeast isolates to the genus level and 77.6% of Gram-positive bacteria, 87.1% of Gram-negative bacteria and 100.0% of yeast isolates to the species level. The overall rate of identification using our protocol was 89.9% (364/405 for genus level identification and 73.1% (296/405 for species level identification. Yeast isolates yielded the lowest confidence scores, which compromised the accuracy of identification. Further optimization of the protein extraction procedure in positive blood cultures is needed to improve the rate of identification.

  17. Ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride as a novel matrix for rapid quantitative and qualitative determination of serum free fatty acids by matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yaping [Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dongdan San Tiao, Beijing 100005 (China); Wang, Yanmin [Department of Clinical Laboratory, Heze Municipal Hospital, Shandong (China); Guo, Shuai; Guo, Yumei; Liu, Hui [Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dongdan San Tiao, Beijing 100005 (China); Li, Zhili, E-mail: lizhili@ibms.pumc.edu.cn [Department of Biophysics and Structural Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dongdan San Tiao, Beijing 100005 (China)

    2013-09-10

    Graphical abstract: -- Highlights: •A novel MALDI matrix for the detection of serum free fatty acids is ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride. •Multiple point internal standard calibration curves were constructed for nine FFAs, respectively, with excellent correlation coefficients between 0.991 and 0.999. •The MALDI-MS approach was used to rapidly differentiate the patients with and without hyperglycemia and healthy controls. -- Abstract: The blood free fatty acids (FFAs), which provide energy to the cell and act as substrates in the synthesis of fats, lipoproteins, liposaccharides, and eicosanoids, involve in a number of important physiological processes. In the present study, matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS) with ammonia-treated N-(1-naphthyl) ethylenediamine dihydrochloride (ATNEDC) as a novel MALDI matrix in a negative ion mode was employed to directly quantify serum FFAs. Multiple point internal standard calibration curves between the concentration ratios of individual fatty acids to internal standard (IS, C{sub 17:0}) versus their corresponding intensity ratios were constructed for C{sub 14:0}, C{sub 16:1}, C{sub 16:0}, C{sub 18:0}, C{sub 18:1}, C{sub 18:2}, C{sub 18:3}, C{sub 20:4}, and C{sub 22:6}, respectively, in their mixture, with correlation coefficients between 0.991 and 0.999 and limits of detection (LODs) between 0.2 and 5.4 μM, along with the linear dynamic range of more than two orders of magnitude. The results indicate that the multiple point internal standard calibration could reduce the impact of ion suppression and improve quantification accuracy in the MALDI mode. The quantitative results of nine FFAs from 339 serum samples, including 161 healthy controls, 118 patients with hyperglycemia and 60 patients without hyperglycemia show that FFAs levels in hyperglycemic patient sera are significantly higher than those in healthy

  18. Laser desorption and time-of-flight mass spectrometry. Fundamentals .Applications; Desorption laser et spectrometrie de masse par temps de vol. Aspects fondamentaux. Applications

    Energy Technology Data Exchange (ETDEWEB)

    Chaurand, P.

    1994-11-01

    Time-of-flight mass spectrometry is a very powerful technique for the analysis of heavy molecular ions (100 000 u and more). The ejection in the gas phase and the ionization of these molecules is now possible through the MALDI technique (Matrix Assisted Laser Desorption Ionization). This technique consists in mixing the heavy molecules to be analysed with a organic matrix which absorbs at the wavelength of the laser. The necessary irradiance are of the order of 10{sup 6} W/cm{sup 2}. In these conditions we have shown that the mass resolutions are optimum and that the relative mass accuracies are of the order of 10{sup -4}. We have also demonstrated that the emission angle of the molecular ions in MALDI depends on the incident angle of the laser light. During the desorption process, the molecular ions are emitted in the opposite direction of the incident laser light. This effect is particularly important for the design of the accelerating stage of the time-of-flight spectrometers. Problems relative to the detection of these heavy molecular ions have been studied in details between 0.5 10{sup 4} m/s and 10{sup 5} m/s. The velocity threshold of the electronic emission is lower than the value of 0.5 10{sup 4} m/s. The relation between the electronic emission and the projectile velocity is complex. Finally, examples on mass identification of C{sub 60} molecules and derivated C{sub 60} are presented. Desorption methods are compared. (author). 32 refs., 34 figs.

  19. Desorption Dynamics, Internal Energies and Imaging of Organic Molecules from Surfaces with Laser Desorption and Vacuum Ultraviolet (VUV) Photoionization

    Energy Technology Data Exchange (ETDEWEB)

    Kostko, Oleg; Takahashi, Lynelle K.; Ahmed, Musahid

    2011-04-05

    There is enormous interest in visualizing the chemical composition of organic material that comprises our world. A convenient method to obtain molecular information with high spatial resolution is imaging mass spectrometry. However, the internal energy deposited within molecules upon transfer to the gas phase from a surface can lead to increased fragmentation and to complications in analysis of mass spectra. Here it is shown that in laser desorption with postionization by tunable vacuum ultraviolet (VUV) radiation, the internal energy gained during laser desorption leads to minimal fragmentation of DNA bases. The internal temperature of laser-desorbed triacontane molecules approaches 670 K, whereas the internal temperature of thymine is 800 K. A synchrotron-based VUV postionization technique for determining translational temperatures reveals that biomolecules have translational temperatures in the range of 216-346 K. The observed low translational temperatures, as well as their decrease with increased desorption laser power is explained by collisional cooling. An example of imaging mass spectrometry on an organic polymer, using laser desorption VUV postionization shows 5 mu m feature details while using a 30 mu m laser spot size and 7 ns duration. Applications of laser desorption postionization to the analysis of cellulose, lignin and humic acids are briefly discussed.

  20. MALDI-TOF-MS 方法检测、鉴定副溶血性弧菌%Detection and identification of Vibrio parahaemolyticus by matrix assisted laser desorption/ionization time of flight mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    龚艳清; 陈信忠; 郭书林; 杨俊萍; 叶妍妍

    2013-01-01

    Objective To establish a rapid method for the detection and identification of Vibrio parahae-molyticus by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Methods In order to verify the stability and repeatability, the reference strain V. parahaemolyticus cultured in different medium or different time and the isolates which had been cryopreserved for a long time were detected by MALDI-TOF-MS. The strains from different sources had also been tested to confirm the accuracy of this method. Results Different medium such as PW, TCBS and Vibrio colored medium and different culturing time interval of 18, 42 or 72 h had little impact for the mass spectrometry of the bacteria. There were no changes for the identification scores of the bacteria which had been preserved for two years using MALDI-TOF-MS. The strains from different countries and regions get the same accurate results. Conclusion MALDI-TOF-MS is a fast and accurate method for the identification of V. parahaemolyticus by MALDI-TOF-MS, and it can be used for routine detection because of its good stability and repeatability.%  目的建立快速检测和鉴定副溶血性弧菌的基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)分析方法。方法采用副溶血性弧菌标准菌株,在不同的培养基或不同的培养时间下进行 MALDI-TOF-MS 检测,同时对长时间冷冻保存的菌株进行检测,以验证该方法的稳定性和重复性。对不同来源的菌株进行检测,以确认方法的准确性。结果用 PW、TCBS 琼脂和弧菌显色培养基等3种培养基,分别培养18、42、72 h 时,对该菌的蛋白质谱图影响很小;保存2年的菌株,其 MALDI-TOF-MS 鉴定分值保持稳定;对来自不同国家和地区的菌株都可以得到同样准确的结果。结论 MALDI-TOF-MS 方法可以快速、准确地鉴定副溶血性弧菌。因其具有很好的稳定性和重复性,可用于副溶血性弧菌的日常检测。

  1. Rapid identification of Nontuberculous Mycobacterium using matrix assisted laser desorption/ionization time of flight mass spectrometry%MALDI-TOF质谱快速鉴定非结核分枝杆菌方法的建立和评价

    Institute of Scientific and Technical Information of China (English)

    周昭彦; 胡必杰; 鲍容; 马坚; 黄声雷; 谢红梅

    2013-01-01

    Objective The purpose of this study was to compare and develop the method for identification of Non-tuberculous Mycobacteria (NTM) using matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS),and evaluate the feasibility,accuracy and repeatability of MALDI-TOF MS to discriminate NTM.Methods Fifteen clinical strains were collected from January to March in 2012 and 68 environmental strains were retrospectively collected from 2011 to 2012.A protocol for sample pre-treatment and protein extraction was developed and utilized it to identify clinical and environmental isolates.The results from 16 s rRNA sequencing were served as control.Results Method A was more effective in protein extraction.Although all the three methods got the same species result,a total of 83 strains belonging to 10 distinct species grown in Middle brook 7H10 media were analyzed.All members of the NTM were identified accurately at the genus level and 80.7% (67/83) of strains could be identified at the species level.Six strains were identified at the complex level.81.9% (68/83) of NTM got high spectral scores.The identification of cultured colony could be completed in 1.5 hours.And it had good reproducibility.Conclusion MALDI-TOF MS can be used as a rapid and accurate method for identification of Mycobacteria in clinical microbiology laboratories,implying its good application prospects.%目的 建立基质辅助激光解析/电离飞行时间质谱(MALDI-TOF MS)鉴定非结核分枝杆菌(NTM)的方法,评估其快速鉴定临床和环境分离NTM的可行性、准确性和重复性.方法 回顾性收集2012年1至3月上海复旦大学附属中山医院微生物实验室临床分离NTM 15株及2011至2012年医院供水系统环境分离NTM 68株.比较3种样本前处理和蛋白抽提方案,采用MALDI-TOFMS技术检测临床和环境分离NTM,并以16S rRNA测序结果为对照.结果 3种方法均鉴定出相同细菌,但方法1的鉴定值高于其他2

  2. Target Plate Material Influence on Fullerene-C60 Laser Desorption/Ionization Efficiency

    Science.gov (United States)

    Zeegers, Guido P.; Günthardt, Barbara F.; Zenobi, Renato

    2016-04-01

    Systematic laser desorption/ionization (LDI) experiments of fullerene-C60 on a wide range of target plate materials were conducted to gain insight into the initial ion formation in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The positive and negative ion signal intensities of precursor, fragment, and cluster ions were monitored, varying both the laser fluence (0-3.53 Jcm-2) and the ion extraction delay time (0-950 ns). The resulting species-specific ion signal intensities are an indication for the ionization mechanisms that contribute to LDI and the time frames in which they operate, providing insight in the (MA)LDI primary ionization. An increasing electrical resistivity of the target plate material increases the fullerene-C60 precursor and fragment anion signal intensity. Inconel 625 and Ti90/Al6/V4, both highly electrically resistive, provide the highest anion signal intensities, exceeding the cation signal intensity by a factor ~1.4 for the latter. We present a mechanism based on transient electrical field strength reduction to explain this trend. Fullerene-C60 cluster anion formation is negligible, which could be due to the high extraction potential. Cluster cations, however, are readily formed, although for high laser fluences, the preferred channel is formation of precursor and fragment cations. Ion signal intensity depends greatly on the choice of substrate material, and careful substrate selection could, therefore, allow for more sensitive (MA)LDI measurements.

  3. Target Plate Material Influence on Fullerene-C60 Laser Desorption/Ionization Efficiency.

    Science.gov (United States)

    Zeegers, Guido P; Günthardt, Barbara F; Zenobi, Renato

    2016-04-01

    Systematic laser desorption/ionization (LDI) experiments of fullerene-C60 on a wide range of target plate materials were conducted to gain insight into the initial ion formation in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The positive and negative ion signal intensities of precursor, fragment, and cluster ions were monitored, varying both the laser fluence (0-3.53 Jcm(-2)) and the ion extraction delay time (0-950 ns). The resulting species-specific ion signal intensities are an indication for the ionization mechanisms that contribute to LDI and the time frames in which they operate, providing insight in the (MA)LDI primary ionization. An increasing electrical resistivity of the target plate material increases the fullerene-C60 precursor and fragment anion signal intensity. Inconel 625 and Ti90/Al6/V4, both highly electrically resistive, provide the highest anion signal intensities, exceeding the cation signal intensity by a factor ~1.4 for the latter. We present a mechanism based on transient electrical field strength reduction to explain this trend. Fullerene-C60 cluster anion formation is negligible, which could be due to the high extraction potential. Cluster cations, however, are readily formed, although for high laser fluences, the preferred channel is formation of precursor and fragment cations. Ion signal intensity depends greatly on the choice of substrate material, and careful substrate selection could, therefore, allow for more sensitive (MA)LDI measurements. Graphical Abstract ᅟ.

  4. Laser assisted hair-removal.

    Science.gov (United States)

    Choudhary, S; Elsaie, M L; Nouri, K

    2009-10-01

    A number of lasers and light devices are now available for the treatment of unwanted hair. The goal of laser hair removal is to damage stem cells in the bulge of the hair follicle by targeting melanin, the endogenous chromophore for laser and light devices utilized to remove hair. The competing chromophores in the skin and hair, oxyhemoglobin and water, have a decreased absorption between 690 nm and 1000 nm, thus making this an ideal range for laser and light sources. Laser hair removal is achieved through follicular unit destruction based on selective photothermolysis. The principle of selective photothermolysis predicts that the thermal injury will be restricted to a given target if there is sufficient selective absorption of light and the pulse duration is shorter than the thermal relaxation time of the target. This review will focus on the mechanisms of laser assisted hair removal and provide an update on the newer technologies emerging in the field of lasers assisted hair removal.

  5. Ambient Femtosecond Laser Vaporization and Nanosecond Laser Desorption Electrospray Ionization Mass Spectrometry

    Science.gov (United States)

    Flanigan, Paul; Levis, Robert

    2014-06-01

    Recent investigations of ambient laser-based transfer of molecules into the gas phase for subsequent mass spectral analysis have undergone a renaissance resulting from the separation of vaporization and ionization events. Here, we seek to provide a snapshot of recent femtosecond (fs) duration laser vaporization and nanosecond (ns) duration laser desorption electrospray ionization mass spectrometry experiments. The former employs pulse durations of explosive formulations, plant tissue phenotypes, and even the identity of the manufacturer of smokeless powders can be determined by multivariate statistics. We also review the more mature field of nanosecond laser desorption for ambient mass spectrometry, covering the wide range of systems analyzed, the need for resonant absorption, and the spatial imaging of complex systems like tissue samples.

  6. Laser desorption ionization and peptide sequencing on laser induced silicon microcolumn arrays

    Science.gov (United States)

    Vertes, Akos [Reston, VA; Chen, Yong [San Diego, CA

    2011-12-27

    The present invention provides a method of producing a laser-patterned silicon surface, especially silicon wafers for use in laser desorption ionization (LDI-MS) (including MALDI-MS and SELDI-MS), devices containing the same, and methods of testing samples employing the same. The surface is prepared by subjecting a silicon substrate to multiple laser shots from a high-power picosecond or femtosecond laser while in a processing environment, e.g., underwater, and generates a remarkable homogenous microcolumn array capable of providing an improved substrate for LDI-MS.

  7. Investigation of thin ZnO layers in view of laser desorption-ionization

    Energy Technology Data Exchange (ETDEWEB)

    Grechnikov, A A; Borodkov, A S [Vernadsky Institute of Geochemistry and Analytical Chemistry, Russian Academy of Sciences, 19 Kosygin Str., 119991 Moscow (Russian Federation); Georgieva, V B [Georgi Nadjakov Institute of Solid State Physics, Bulgarian Academy of Sciences, 72 Tzarigradsko Chaussee, 1784 Sofia (Bulgaria); Alimpiev, S S; Nikiforov, S M; Simanovsky, Ya O [General Physics Institute, Russian Academy of Sciences, 38 Vavilov Str., 119991 Moscow (Russian Federation); Dimova-Malinovska, D; Angelov, O I, E-mail: lazarova@issp.bas.b [Laboratory for Solar Energy and New Energy Sources, Bulgarian Academy of Sciences, 72 Tzarigradsko Chaussee, 1784 Sofia (Bulgaria)

    2010-04-01

    Thin zinc oxide films (ZnO) were developed as a matrix-free platform for surface assisted laser desorption-ionization (SALDI) time-of-flight mass spectrometry. The ZnO films were deposited by RF magnetron sputtering of ZnO ceramic targets in Ar atmospheres on monocrystalline silicon. The generation under UV (355 nm) laser irradiation of positive ions of atenolol, reserpine and gramicidin S from the ZnO layers deposited was studied. All analytes tested were detected as protonated molecules with no or very structure-specific fragmentation. The mass spectra obtained showed low levels of chemical background noise. All ZnO films studied exhibited high stability and good reproducibility. The detection limits for test analytes are in the 10 femtomol range.

  8. Novel Galvanic Nanostructures of Ag and Pd for Efficient Laser Desorption/Ionization of Low Molecular Weight Compounds

    Science.gov (United States)

    Silina, Yuliya E.; Meier, Florian; Nebolsin, Valeriy A.; Koch, Marcus; Volmer, Dietrich A.

    2014-05-01

    A simple approach for synthesis of palladium and silver nanostructures with readily adjustable morphologies was developed using galvanic electrochemical deposition, for application to surface-assisted laser desorption/ionization (SALDI) of small biological molecules. A range of fatty acids, triglycerides, carbohydrates, and antibiotics were investigated to assess the performance of the new materials. Intense analyte cations were generated from the galvanic surfaces upon UV laser irradiation such as potassium adducts for a film thickness 120 nm. Possible laser desorption/ionization mechanisms of these galvanic structures are discussed. The films exhibited self-organizing abilities and adjustable morphologies by changing electrochemical parameters. They did not require any stabilizing agents and were inexpensive and very easy to produce. SALDI analysis showed that the materials were stable under ambient conditions and analytical results with excellent measurement reproducibility and detection sensitivity similar to MALDI were obtained. Finally, we applied the galvanic surfaces to fast screening of natural oils with minimum sample preparation.

  9. Laser Desorption Ionization Quadrupole Ion Trap Time-of-Flight Mass Spectrometry of Au m Fe n +/- Clusters Generated from Gold-Iron Nanoparticles and their Giant Nanoflowers. Electrochemical and/or Plasma Assisted Synthesis

    Science.gov (United States)

    Mawale, Ravi Madhukar; Ausekar, Mayuri Vilas; Pavliňák, David; Galmiz, Oleksandr; Kubáček, Pavel; Havel, Josef

    2017-02-01

    Gold nanoparticles (NP) with average diameter 100 nm synthesized from tetrachloroauric acid solution using stainless steel as a reducing agent were found to contain iron. Applying simultaneously high frequency (HF) plasma discharge in solution during the electrochemical reduction, giant gold-iron nanoflowers with average size 1000-5000 nm were formed. Scanning electron microscopy (SEM) shows the morphology of the nanopowders produced as polygonal yet nearly spherical, whereas iron content in both products determined by energy dispersive X-ray analysis (EDX) was found to be at 2.5 at. %. Laser desorption ionization (LDI) of both nanomaterials and mass spectrometric analysis show the formation of Au m Fe n +/- ( m = 1-35; n = 1-3) clusters. Structure of few selected clusters in neutral or monocharged forms were computed by density functional theory (DFT) calculations and it was found that typical distances of an iron nucleus from adjacent gold nuclei lie in the interval 2.5 to 2.7 Å. Synthetized Au-Fe nanoparticles were found stable for at least 2 mo at room temperature (even in aqueous solution) without any stabilizing agent. Produced Au-Fe nanoparticles in combination with standard MALDI matrices enhance ionization of peptides and might find use in nanomedicine.

  10. Site Specificity in Femtosecond Laser Desorption of Neutral H Atoms from Graphite(0001)

    DEFF Research Database (Denmark)

    Frigge, R.; Hoger, T.; Siemer, B.;

    2010-01-01

    Femtosecond laser excitation and density functional theory reveal site and vibrational state specificity in neutral atomic hydrogen desorption from graphite induced by multiple electronic transitions. Multimodal velocity distributions witness the participation of ortho and para pair states...... of chemisorbed hydrogen in the desorption process. Very slow velocities of 700 and 400  ms-1 for H and D atoms are associated with the desorption out of the highest vibrational state of a barrierless potential....

  11. Electronically driven adsorbate excitation mechanism in femtosecond-pulse laser desorption

    DEFF Research Database (Denmark)

    Brandbyge, Mads; Hedegård, Per; Heinz, T. F.

    1995-01-01

    Femtosecond-pulse laser desorption is a process in which desorption is driven by a subpicosecond temperature pulse of order 5000 K in the substrate-adsorbate electron system, whose energy is transferred into the adsorbate center-of-mass degrees of freedom by a direct coupling mechanism. We presen...

  12. Study on impurity desorption induced by femtosecond pulse laser based on a stochastic process model

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    With the advantages on non-equilibrium heating and desorption induced by electronic transition, the femtosecond pulse laser introduces a new way for solving the problem of impurity pollution adsorbed on a solid thin film in micro-electro-mechanical systems (MEMS). A model based on stochastic processes was established for stimulated desorption induced by the femtosecond pulse laser to interpret the interaction of the optically excited hot electrons with the adsorbed molecules in a metal substrate. Numerical simulation results reveal a time-dependent desorption probability of adsorbed molecules and indicate that how key parameters of femtosecond pulse laser, such as incident laser energy flux, pulse duration, and wavelength of pulse, have a great effect on the desorption probability.

  13. Ammonium Ion Exchanged Zeolite for Laser Desorption/Ionization Mass Spectrometry of Phosphorylated Peptides

    Directory of Open Access Journals (Sweden)

    Mengrui Yang

    2015-01-01

    Full Text Available α-Cyano-4-hydroxycinnamic acid (CHCA, an organic matrix molecule for matrix-assisted laser desorption/ionization mass spectrometry, was adsorbed to NH4+-type zeolite surface, and this new matrix was used for the detection of low-molecular-weight compounds. It was found that this matrix could simplify the mass spectrum in the low-molecular-weight region and prevent interference from fragments and alkali metal ion adducted species. CHCA adsorbed to NH4+-type ZSM5 zeolite (CHCA/NH4ZSM5 was used to measure atropine and aconitine, two toxic alkaloids in plants. In addition, CHCA/NH4ZSM5 enabled us to detect phosphorylated peptides; peaks of the protonated peptides had higher intensities than the peaks observed using CHCA only.

  14. Laser Desorption Supersonic Jet Spectroscopy of Hydrated Tyrosine

    Science.gov (United States)

    Oba, Hikari; Shimozono, Yoko; Ishiuchi, Shun-Ichi; Fujii, Masaaki; Carcabal, Pierre

    2013-06-01

    The structure of tyrosine (tyr) consists of amino-acid chain and phenol, and it has roughly two possible binding sites for water, amino-acid site and phenolic OH site. Investigating how water molecule binds to tyr will give fundamental information for hydrations of peptide and protein. Resonance enhanced multi photon ionization (REMPI) spectrum of tyr-water 1:1 cluster has already been reported by de Vries and co-workers, however, no analysis on the hydrated structures has been reported. In the REMPI spectrum, two clusters of bands are observed; one appears at ˜35600 cm^{-1} energy region which is the almost same with 0-0 transitions of tyr monomer, and another is observed at ˜300 cm^{-1} lower than the former. Based on the electronic transition energy of phenylalanine and the hydrated clusters, the former is expected to be derived from a structure that water binds to amino acid site. On the other hand, it is plausibly predicted that the latter originates from a structure that water binds to phenolic OH group, because the electronic transition of mono hydrated phenol is ˜300 cm^{-1} red-shifted from the monomer. We applied IR dip spectroscopy which can measure conformer selective IR spectra to the tyr-(H_{2}O)_{1} clusters by using laser desorption supersonic jet technique to confirm the assignments. Especially in the phenolic OH bound isomer, it was found that the intra molecular hydrogen bond within amino-acid chain, which is far from the water molecule and cannot interact directly with each other, is strengthened by the hydration. A. Abio-Riziq et al., J. Phys. Chem. A, 115, 6077 (2011). Y. Shimozono, et al., Phys. Chem. Chem. Phys., (2013) DOI: 10.1039/c3cp43573c. T. Ebata et al., Phys. Chem. Chem. Phys., 8, 4783 (2006). T. Watanabe et al., J. Chem. Phys., 105, 408 (1996).

  15. Laser-assisted fabrication of materials

    CERN Document Server

    Manna, Indranil

    2013-01-01

    Laser assisted fabrication involves shaping of materials using laser as a source of heat. It can be achieved by removal of materials (laser assisted cutting, drilling, etc.), deformation (bending, extrusion), joining (welding, soldering) and addition of materials (surface cladding or direct laser cladding). This book on ´Laser assisted Fabrication’ is aimed at developing in-depth engineering concepts on various laser assisted macro and micro-fabrication techniques with the focus on application and a review of the engineering background of different micro/macro-fabrication techniques, thermal history of the treated zone and microstructural development and evolution of properties of the treated zone.

  16. 基质辅助激光解析电离飞行时间质谱快速鉴定耐甲氧西林金黄色葡萄球菌%Rapid method study of methicillin-resistant Staphylococcus aureus identified by Matrix-Assisted laser desorption Ionization-Time of flight mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    孙宗科; 张伟; 陈西平

    2011-01-01

    目的 研究基质辅助激光解析电离飞行时间质谱(Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry,MALDI-TOF-Ms)用于快速检测鉴定金黄色葡萄球菌及其耐药性的可行性.方法 用MALDI-TOF-MS和API Staph系统鉴定了40株临床分离的葡萄球菌,用Kirby-Bauer法检测金黄色葡萄球菌的耐药性.结果 40株临床分离的葡萄球菌中34株细菌用MALDI-TOF-MS鉴定为金黄色葡萄球菌,6株为其他葡萄球菌.与API Staph系统鉴定结果一致.Kirby-Bauer法耐药性检测显示其中11株为耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA),23株为甲氧西林敏感金黄色葡萄球菌(methicillin-susceptible Staphylococcus aureus,MSSA),34株金黄色葡萄球菌用MALDI-TOF-MS鉴定时聚类为2类,结果与耐药性检测结果一致.结论 结果提示MALDI-TOF-MS可以用于MRSA的快速鉴定.%Objective To establish the method on rapid identification of Staphylococcus aureus and the performance of methicillin-resistance. Methods 40 isolation strains of staphylococcal were analyzed by MatrixAssisted Laser Desorption Ionization-Time of Flight Mass Spectrometry ( MALDI-TOF-MS ) and APl Staph Ident system, and the performance of methicillin-resistance was tested by Disk Diffusion Susceptibility Testing. Results 34 isolation strains were identified by MALDI-TOF-MS as Staphylococcus aureus, and the result was uniform to the APl Staph Ident system. 11 isolation strains were identified as methicillin-resistant Staphylococcus aureus (MRSA)and 23 isolation strains were methicillin-susceptible to Staphylococcus aureus ( MSSA ). All the 34 isolation strains were clustered to 2 species identified by MALDI-TOF-MS. The result was uniform to the Disk Diffusion Susceptibility Testing. Conclusion These results demonstrated that MALDI-TOF-MS could be a rapid technique for identification and discrimination of MRSA.

  17. Laser desorption/ionization mass spectrometry for direct profiling and imaging of small molecules from raw biological materials

    Energy Technology Data Exchange (ETDEWEB)

    Cha, Sangwon [Iowa State Univ., Ames, IA (United States)

    2008-01-01

    Matrix-assisted laser desorption/ionization(MALDI) mass spectrometry(MS) has been widely used for analysis of biological molecules, especially macromolecules such as proteins. However, MALDI MS has a problem in small molecule (less than 1 kDa) analysis because of the signal saturation by organic matrixes in the low mass region. In imaging MS (IMS), inhomogeneous surface formation due to the co-crystallization process by organic MALDI matrixes limits the spatial resolution of the mass spectral image. Therefore, to make laser desorption/ionization (LDI) MS more suitable for mass spectral profiling and imaging of small molecules directly from raw biological tissues, LDI MS protocols with various alternative assisting materials were developed and applied to many biological systems of interest. Colloidal graphite was used as a matrix for IMS of small molecules for the first time and methodologies for analyses of small metabolites in rat brain tissues, fruits, and plant tissues were developed. With rat brain tissues, the signal enhancement for cerebroside species by colloidal graphite was observed and images of cerebrosides were successfully generated by IMS. In addition, separation of isobaric lipid ions was performed by imaging tandem MS. Directly from Arabidopsis flowers, flavonoids were successfully profiled and heterogeneous distribution of flavonoids in petals was observed for the first time by graphite-assisted LDI(GALDI) IMS.

  18. Chemical characterization of latent fingerprints by matrix-assisted laser desorption ionization, time-of-flight secondary ion mass spectrometry, mega electron volt secondary mass spectrometry, gas chromatography/mass spectrometry, X-ray photoelectron spectroscopy, and attenuated total reflection Fourier transform infrared spectroscopic imaging: an intercomparison.

    Science.gov (United States)

    Bailey, Melanie J; Bright, Nicholas J; Croxton, Ruth S; Francese, Simona; Ferguson, Leesa S; Hinder, Stephen; Jickells, Sue; Jones, Benjamin J; Jones, Brian N; Kazarian, Sergei G; Ojeda, Jesus J; Webb, Roger P; Wolstenholme, Rosalind; Bleay, Stephen

    2012-10-16

    The first analytical intercomparison of fingerprint residue using equivalent samples of latent fingerprint residue and characterized by a suite of relevant techniques is presented. This work has never been undertaken, presumably due to the perishable nature of fingerprint residue, the lack of fingerprint standards, and the intradonor variability, which impacts sample reproducibility. For the first time, time-of-flight secondary ion mass spectrometry, high-energy secondary ion mass spectrometry, and X-ray photoelectron spectroscopy are used to target endogenous compounds in fingerprints and a method is presented for establishing their relative abundance in fingerprint residue. Comparison of the newer techniques with the more established gas chromatography/mass spectrometry and attenuated total reflection Fourier transform infrared spectroscopic imaging shows good agreement between the methods, with each method detecting repeatable differences between the donors, with the exception of matrix-assisted laser desorption ionization, for which quantitative analysis has not yet been established. We further comment on the sensitivity, selectivity, and practicability of each of the methods for use in future police casework or academic research.

  19. Rapid detection of B2-ST131 clonal group of extended-spectrum β-lactamase-producing Escherichia coli by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry: discovery of a peculiar amino acid substitution in B2-ST131 clonal group.

    Science.gov (United States)

    Nakamura, Akihiro; Komatsu, Masaru; Kondo, Akira; Ohno, Yuki; Kohno, Hisashi; Nakamura, Fumihiko; Matsuo, Shuji; Ohnuma, Kenichiro; Hatano, Naoya; Kawano, Seiji

    2015-11-01

    One reason for the spread of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli worldwide is the global pandemic of the B2-ST131 clonal group. We searched for the specific biomarker peaks to distinguish between the B2-ST131 clonal group and other sequence type (ST) clonal groups isolated from clinical specimens obtained in our hospital. Biomarker peaks at m/z 7650 in the B2-ST131 group (sensitivity of 100% and specificity of 89.7%) and m/z 7707 in the other ST clonal groups showed the highest discrimination abilities. We further verified reproducibility against other Japanese clinical isolates obtained in another area of Japan. Differences between the molecular mass at the 7650m/z and 7707m/z peaks indicated an E34A amino acid substitution by proteomic and genomic analysis. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry rapidly and simply identified the B2-ST131 clonal group in routine examinations and will allow for adequate empirical therapy and the possibility to control both hospital infections and the global pandemic.

  20. Comparison of Biolog GEN III MicroStation semi-automated bacterial identification system with matrix-assisted laser desorption ionization-time of flight mass spectrometry and 16S ribosomal RNA gene sequencing for the identification of bacteria of veterinary interest.

    Science.gov (United States)

    Wragg, P; Randall, L; Whatmore, A M

    2014-10-01

    Recent advances in phenotypic and chemotaxonomic methods have improved the ability of systems to resolve bacterial identities at the species level. Key to the effective use of these systems is the ability to draw upon databases which can be augmented with new data gleaned from atypical or novel isolates. In this study we compared the performance of the Biolog GEN III identification system (hereafter, GEN III) with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequencing in the identification of isolates of veterinary interest. The use of strains that had proven more difficult to identify by routine methods was designed to test the systems' abilities at the extremes of their performance range. Over an 18month period, 100 strains were analysed by all three methods. To highlight the importance of identification to species level, a weighted scoring system was devised to differentiate the capacity to identify at genus and species levels. The overall relative weighted scores were 0.869:0.781:0.769, achieved by 16S rRNA gene sequencing, GEN III and MALDI-TOF MS respectively, when compared to the 'gold standard'. Performance to the genus level was significantly better using 16S rRNA gene sequencing; however, performance to the species level was similar for all three systems.

  1. Carbon nanodots as a matrix for the analysis of low-molecular-weight molecules in both positive- and negative-ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantification of glucose and uric acid in real samples.

    Science.gov (United States)

    Chen, Suming; Zheng, Huzhi; Wang, Jianing; Hou, Jian; He, Qing; Liu, Huihui; Xiong, Caiqiao; Kong, Xianglei; Nie, Zongxiu

    2013-07-16

    Carbon nanodots were applied for the first time as a new matrix for the analysis of low-molecular-weight compounds by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in both positive- and negative-ion modes. A wide range of small molecules including amino acids, peptides, fatty acids, as well as β-agonists and neutral oligosaccharides were analyzed by MALDI MS with carbon nanodots as the matrix, and the lowest 0.2 fmol limits-of-detection were obtained for octadecanoic acid. Clear sodium and potassium adducts and deprotonated signals were produced in positive- and negative-ion modes. Furthermore, the glucose and uric acid in real samples were quantitatively determined by the internal standard method with the linear range of 0.5-9 mM and 0.1-1.8 mM (R(2) > 0.999), respectively. This work gives new insight into the application of carbon nanodots and provides a general approach for rapid analysis of low-molecular-weight compounds.

  2. Non-traditional applications of laser desorption/ionization mass spectrometry

    Science.gov (United States)

    McAlpin, Casey R.

    Seven studies were carried out using laser desorption/ionization mass spectrometry (LDI MS) to develop enhanced methodologies for a variety of analyte systems by investigating analyte chemistries, ionization processes, and elimination of spectral interferences. Applications of LDI and matrix assisted laser/desorption/ionization (MALDI) have been previously limited by poorly understood ionization phenomena, and spectral interferences from matrices. Matrix assisted laser desorption ionization MS is well suited to the analysis of proteins. However, the proteins associated with bacteriophages often form complexes which are too massive for detection with a standard MALDI mass spectrometer. As such, methodologies for pretreatment of these samples are discussed in detail in the first chapter. Pretreatment of bacteriophage samples with reducing agents disrupted disulfide linkages and allowed enhanced detection of bacteriophage proteins. The second chapter focuses on the use of MALDI MS for lipid compounds whose molecular mass is significantly less than the proteins for which MALDI is most often applied. The use of MALDI MS for lipid analysis presented unique challenges such as matrix interference and differential ionization efficiencies. It was observed that optimization of the matrix system, and addition of cationization reagents mitigated these challenges and resulted in an enhanced methodology for MALDI MS of lipids. One of the challenges commonly encountered in efforts to expand MALDI MS applications is as previously mentioned interferences introduced by organic matrix molecules. The third chapter focuses on the development of a novel inorganic matrix replacement system called metal oxide laser ionization mass spectrometry (MOLI MS). In contrast to other matrix replacements, considerable effort was devoted to elucidating the ionization mechanism. It was shown that chemisorption of analytes to the metal oxide surface produced acidic adsorbed species which then

  3. Rhabdomyolysis After Cosmetic Laser-Assisted Liposuction.

    Science.gov (United States)

    Shin, Jin-yong; Chang, Hak

    2015-08-01

    A 34-year-old-female patient visited our emergency room for symptoms of disturbance of urination and shortness of breath. She was diagnosed with rhabdomyolysis with acute kidney injury after laser-assisted liposuction and required hemodialysis. Although laser-assisted liposuction is a well-used procedure, it can cause local complications, such as burns and skin irregularities, as well as systemic complications, such as infection, fever, and emboli. However, laser-assisted, liposuction-induced rhabdomyolysis has not been reported. Repetitive exercises, trauma, and crush injury are the major causes of rhabdomyolysis. In this study, a unique case of rhabdomyolysis that developed after laser-assisted liposuction is reported.

  4. Laser Infrared Desorption Spectroscopy to Detect Complex Organic Molecules on Icy Planetary Surfaces

    Science.gov (United States)

    Sollit, Luke S.; Beegle, Luther W.

    2008-01-01

    Laser Desorption-Infrared Spectroscopy (LD-IR) uses an IR laser pulse to desorb surface materials while a spectrometer measures the emission spectrum of the desorbed materials (Figure 1). In this example, laser desorption operates by having the incident laser energy absorbed by near surface material (10 microns in depth). This desorption produces a plume that exists in an excited state at elevated temperatures. A natural analog for this phenomenon can be observed when comets approach the sun and become active and individual molecular emission spectra can be observed in the IR [1,2,3,4,5]. When this occurs in comets, the same species that initially emit radiation down to the ground state are free to absorb it, reducing the amount of detectable emission features. The nature of our technique results in absorption not occurring, because the laser pulse could easily be moved away form the initial desorption plume, and still have better spatial resolution then reflectance spectroscopy. In reflectance spectroscopy, trace components have a relatively weak signal when compared to the entire active nature of the surface. With LDIR, the emission spectrum is used to identify and analyze surface materials.

  5. 2,5-Dihydroxybenzoic acid: laser desorption/ionisation as a function of elevated temperature

    Science.gov (United States)

    Wallace, W. E.; Arnould, M. A.; Knochenmuss, R.

    2005-03-01

    The temperature dependence of laser desorption/ionization (LDI) ion yields has been measured for 2,5-dihydroxybenzoic acid (2,5-DHB) single crystals from room temperature to 160 °C using time-of-flight (TOF) mass spectrometry. A steep rise in ion production occurs at 90 °C, achieving a maximum at 120 °C, then decreases sharply to a minimum at 140 °C, and returns to a second maximum at 150 °C. Above 160 °C, useful information could not be obtained because of rapid volatilization of the sample into the vacuum. The overall trend in ion production, but not some of the details, is well described by a recent two-step theory of the laser desorption/ionization process, which takes into account the temperature-dependent effects of plume expansion. Measuring the background vacuum composition with a quadrupole mass spectrometer residual gas analyzer (RGA) showed an increase in thermal desorption of 2,5-DHB starting at 90 °C and maximizing at 130 °C. The increased neutral production by thermal desorption is believed to be the cause of the decrease in LDI ion production due to reduced pooling probabilities for laser-excited 2,5-DHB molecules. Thermal dehydration, condensation, and decarboxylation increase the volume of gas released at high temperatures which also serve to decrease LDI ion production at elevated temperatures. Lastly, to confirm the mass spectrometry results, the thermal desorption of 2,5-DHB single crystals under vacuum was measured using a quartz-crystal microbalance (QCM). The onset of desorption was found to occur at 90 °C and the maximum desorption rate was found at 135 °C.

  6. Laser Desorption Mass Spectrometry for High Throughput DNA Analysis and Its Applications

    Energy Technology Data Exchange (ETDEWEB)

    Allman, S.L.; Chen, C.H.; Golovlev, V.V.; Isola, N.R.; Matteson, K.J.; Potter, N.T.; Taranenko, N.I.

    1999-01-23

    Laser desorption mass spectrometry (LDMS) has been developed for DNA sequencing, disease diagnosis, and DNA Fingerprinting for forensic applications. With LDMS, the speed of DNA analysis can be much faster than conventional gel electrophoresis. No dye or radioactive tagging to DNA segments for detection is needed. LDMS is emerging as a new alternative technology for DNA analysis.

  7. Matrix-assisted laser desorption ionization-time of flight mass spectrometry for rapid identification of fungi%基质辅助激光解吸电离飞行时间质谱在真菌快速鉴定中的应用

    Institute of Scientific and Technical Information of China (English)

    王欢; 曲芬

    2016-01-01

    真菌是广泛分布于自然界的真核生物,对人类具有致病性的约300种。不同种真菌的生长特性、临床特点及耐药性不同。真菌生长缓慢,传统的培养鉴定方法所需时间较长,且日益多样化的真菌,使鉴定难度不断增加,这些均限制了临床的早期诊断和针对性治疗。基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionization-time of flight mass spectrometry, MALDI-TOF MS)是一种新兴的诊断技术,可以通过直接检测生物标志物(蛋白)来鉴定病毒、细菌、分枝杆菌等,具有操作简便、快速、准确率高、成本低的特点。本文对MALDI-TOF MS在真菌鉴定中的应用进行综述,发现其对酵母样真菌的正确鉴定率可达到94%以上,丝状真菌正确鉴定率也可达到89%,可满足临床实验室鉴定真菌的需求。%Fungi are widely distributed in nature, and about 300 species of fungi are pathogenic to humans with different growth characteristics, clinical features and drug resistance. As fungi are slowly growing microorganisms, the culture and identification of fungi by conventional methods are time-consuming. In addition, the increasing diversity of fungal pathogens makes the identification more difficult. All these restrict the early diagnosis and targeted therapies. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is an emerging diagnostic technique in recent years, which can identify viruses, bacteria, mycobacteria and so on through direct detection of biomarkers (proteins). It is a simple and fast diagnostic method with high accuracy and low cost. This review focuses on the identification of fungi by MALDI-TOF MS. Studies have shown that correct identification by MALDI-TOF MS is observed in more than 94% of yeast-like fungi, and 89% of filamentous fungi. Therefore, MALDI-TOF MS proves to meet the demand for the identification of fungi in

  8. Exploration and research of rapidly identifying escherichia coli and detecting ESBLs strains by matrix-assisted laser desorption ionization-time of flight mass spectrometry%MALDI-TOF MS方法快速鉴定大肠杆菌及ESBLs菌株检测探索研究

    Institute of Scientific and Technical Information of China (English)

    战晓微; 杜影; 王宏伟; 吴远高; 郑秋月

    2015-01-01

    Objective The strains of Escherichia coli was tested by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). ESBLs strains were distinguished, and different type ESBLs strains was differentiated by clustering analysis. Feasibility of identification was explored by MALDI-TOF MS for E.coli and ESBLs strains. Methods Peptide mass fingerprints were obtained via collecting mass spectrometric data of 47 E.coli. Their species were identified by biotyper sortware, and drug resistance results were studied. ESBLs strains of E.coli were researched by clustering analysis. Feasibility of identification was confirmed by MALDI-TOF MS for E.coli and ESBLs strains. Results The identification results were coincident with conventional method and PCR method. There were 9 ESBLs stains in 10 E.coli that were prompted for ESBLs strains by MALDI-TOF MS. It had good distinguishing capability for OXA type ESBLs E.coli by cluster analysis. Conclusion E.coli strains could be rapidly, accurately identified by MALDI-TOF MS, and there was a certain screening capacity for ESBLs strains.%目的:研以基质辅助激光解析电离飞行时间质谱(matrix assisted laser desorption ionization-time of flight mass spectrometry, MALDI-TOF MS)方法检测大肠杆菌,并利用MALDI-TOF MS方法鉴别ESBLs菌株及聚类分析法区分不同分型的ESBLs菌株,探索MALDI-TOF MS方法鉴定大肠杆菌及ESBLs菌株的可行性。方法利用MALDI-TOF MS采集47株大肠杆菌分离株的质谱数据,生成肽指纹图谱,并利用biotyper软件进行菌种鉴定,同时研究其耐药性鉴定结果,并利用聚类分析法研究大肠杆菌 ESBLs 菌株,最终确定MALDI-TOF MS对大肠杆菌菌种鉴定及ESBLs菌株鉴别的可行性。结果 MALDI-TOF MS方法鉴定大肠杆菌的结果与传统方法及PCR方法检测结果均符合,经MALDI-TOF MS鉴定方法提示为ESBLs菌株的10株大肠杆菌中有9株为 ESBLs 菌株,且聚类分析法对 OXA

  9. 克罗诺杆菌MALDI-TOF-MS数据库的建立及应用%Establishment and Application of an Analytical Database for Cronobacter spp.by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    赵贵明; 刘洋; 陈颖; 杨海荣; 赵勇胜; 王娉

    2014-01-01

    应用基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionisation time-of-flight mass spectrometry,MALDI-TOF-MS)技术建立了克罗诺杆菌MALDI-TOF-MS数据库,用于该菌属内种和亚种的高通量鉴定及菌株分型.在优化培养条件、样品处理方法及确定MALDI-TOF-MS蛋白质指纹图谱采集参数的基础上,将采集的8种参考菌株蛋白质质谱数据通过Biotyper软件构建克罗诺杆菌的MALDI-TOF-MS数据库,再用相近肠杆菌及该属分离株验证数据库的准确性.结果表明:应用建立的克罗诺杆菌的MALDI-TOF-MS数据库对135株克罗诺杆菌分离株进行分析,鉴定分值均不小于2.0,达到了种水平鉴定要求,通过聚类分析,可进一步分型.构建的MALDI-TOF-MS数据库为克罗诺杆菌高通量鉴定与分型提供了一种新的手段.

  10. Modified 16S-23S rRNA intergenic region restriction endonuclease analysis for species identification of Enterococcus strains isolated from pigs, compared with identification using classical methods and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Nowakiewicz, Aneta; Ziółkowska, Grażyna; Zięba, Przemysław; Trościańczyk, Aleksandra; Banach, Tomasz; Kowalski, Cezary

    2015-03-01

    Fast and reliable identification of bacteria to at least the species level is currently the basis for correct diagnosis and appropriate treatment of infections. This is particularly important in the case of bacteria of the genus Enterococcus, whose resistance profile is often correlated with their species (e.g. resistance to vancomycin). In this study, we evaluated restriction endonuclease analysis of the 16S-23S rRNA gene intergenic transcribed spacer (ITS) region for species identification of Enterococcus. The utility of the method was compared with that of phenotypic methods [biochemical profile evaluation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)]. Identification was based on 21 Enterococcus reference strains, of the species E. faecalis, E. faecium, E. hirae, E. durans, E. casseliflavus, E. gallinarum, E. avium, E. cecorum and E. columbae, and 47 Enterococcus field strains isolated from pigs. Restriction endonuclease analysis of the ITS-PCR product using HinfI, RsaI and MboI, in the order specified, enabled species differentiation of the Enterococcus reference and field strains, and in the case of the latter, the results of species identification were identical (47/47) to those obtained by MALDI-TOF MS. Moreover, as a result of digestion with MboI, a unique restriction profile was also obtained for the strains (3/3) identified by MALDI-TOF MS as E. thailandicus. In our opinion, restriction endonuclease analysis of the 16S-23S rRNA gene ITS region of Enterococcus may be a simple and relatively fast (less than 4 h) alternative method for identifying the species occurring most frequently in humans and animals.

  11. Multidrug-Resistant Candida auris Misidentified as Candida haemulonii: Characterization by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry and DNA Sequencing and Its Antifungal Susceptibility Profile Variability by Vitek 2, CLSI Broth Microdilution, and Etest Method.

    Science.gov (United States)

    Kathuria, Shallu; Singh, Pradeep K; Sharma, Cheshta; Prakash, Anupam; Masih, Aradhana; Kumar, Anil; Meis, Jacques F; Chowdhary, Anuradha

    2015-06-01

    Candida auris is a multidrug-resistant yeast that causes a wide spectrum of infections, especially in intensive care settings. We investigated C. auris prevalence among 102 clinical isolates previously identified as Candida haemulonii or Candida famata by the Vitek 2 system. Internal transcribed spacer region (ITS) sequencing confirmed 88.2% of the isolates as C. auris, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) easily separated all related species, viz., C. auris (n = 90), C. haemulonii (n = 6), C. haemulonii var. vulnera (n = 1), and Candida duobushaemulonii (n = 5). The in vitro antifungal susceptibility was determined using CLSI broth microdilution (CLSI-BMD), the Vitek 2 antifungal susceptibility test, and the Etest method. C. auris isolates revealed uniformly elevated fluconazole MICs (MIC50, 64 μg/ml), and an alarming percentage of isolates (37%) exhibited elevated caspofungin MICs by CLSI-BMD. Notably, 34% of C. auris isolates had coexisting elevated MICs (≥2 μg/ml) for both fluconazole and voriconazole, and 10% of the isolates had elevated coexisting MICs (≥2 μg/ml) to two additional azoles, i.e., posaconazole and isavuconazole. In contrast to reduced amphotericin B MICs by CLSI-BMD (MIC50, 1 μg/ml) for C. auris, elevated MICs were noted by Vitek 2 (MIC50, 8 μg/ml), which were statistically significant. Candida auris remains an unnoticed pathogen in routine microbiology laboratories, as 90% of the isolates characterized by commercial identification systems are misidentified as C. haemulonii. MALDI-TOF MS proved to be a more robust diagnostic technique for rapid identification of C. auris. Considering that misleading elevated MICs of amphotericin B by the Vitek AST-YS07 card may lead to the selection of inappropriate therapy, a cautionary approach is recommended for laboratories relying on commercial systems for identification and antifungal susceptibility testing of rare yeasts.

  12. Multidrug-Resistant Candida auris Misidentified as Candida haemulonii: Characterization by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry and DNA Sequencing and Its Antifungal Susceptibility Profile Variability by Vitek 2, CLSI Broth Microdilution, and Etest Method

    Science.gov (United States)

    Kathuria, Shallu; Singh, Pradeep K.; Sharma, Cheshta; Prakash, Anupam; Masih, Aradhana; Kumar, Anil

    2015-01-01

    Candida auris is a multidrug-resistant yeast that causes a wide spectrum of infections, especially in intensive care settings. We investigated C. auris prevalence among 102 clinical isolates previously identified as Candida haemulonii or Candida famata by the Vitek 2 system. Internal transcribed spacer region (ITS) sequencing confirmed 88.2% of the isolates as C. auris, and matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) easily separated all related species, viz., C. auris (n = 90), C. haemulonii (n = 6), C. haemulonii var. vulnera (n = 1), and Candida duobushaemulonii (n = 5). The in vitro antifungal susceptibility was determined using CLSI broth microdilution (CLSI-BMD), the Vitek 2 antifungal susceptibility test, and the Etest method. C. auris isolates revealed uniformly elevated fluconazole MICs (MIC50, 64 μg/ml), and an alarming percentage of isolates (37%) exhibited elevated caspofungin MICs by CLSI-BMD. Notably, 34% of C. auris isolates had coexisting elevated MICs (≥2 μg/ml) for both fluconazole and voriconazole, and 10% of the isolates had elevated coexisting MICs (≥2 μg/ml) to two additional azoles, i.e., posaconazole and isavuconazole. In contrast to reduced amphotericin B MICs by CLSI-BMD (MIC50, 1 μg/ml) for C. auris, elevated MICs were noted by Vitek 2 (MIC50, 8 μg/ml), which were statistically significant. Candida auris remains an unnoticed pathogen in routine microbiology laboratories, as 90% of the isolates characterized by commercial identification systems are misidentified as C. haemulonii. MALDI-TOF MS proved to be a more robust diagnostic technique for rapid identification of C. auris. Considering that misleading elevated MICs of amphotericin B by the Vitek AST-YS07 card may lead to the selection of inappropriate therapy, a cautionary approach is recommended for laboratories relying on commercial systems for identification and antifungal susceptibility testing of rare yeasts. PMID:25809970

  13. Laser mass spectrometry for biopolymers

    Energy Technology Data Exchange (ETDEWEB)

    Tang, K. (Vanderbilt Univ., Nashville, TN (United States)); Allman, S.L.; Chen, C.H. (Oak Ridge National Lab., TN (United States))

    1992-01-01

    Various matrix materials were used for laser desorption of biological molecules which include large polypeptides and oligonucleotides. Both matrix assisted laser desorption ionization (MALDI) and matrix assisted desorption with post-ionization (MADPI) have been used. Detection sensitivity of femtomole of both oligomer and protein has been achieved. Both positive and negative ions were observed with little fragmentation.

  14. Laser mass spectrometry for biopolymers

    Energy Technology Data Exchange (ETDEWEB)

    Tang, K. [Vanderbilt Univ., Nashville, TN (United States); Allman, S.L.; Chen, C.H. [Oak Ridge National Lab., TN (United States)

    1992-07-01

    Various matrix materials were used for laser desorption of biological molecules which include large polypeptides and oligonucleotides. Both matrix assisted laser desorption ionization (MALDI) and matrix assisted desorption with post-ionization (MADPI) have been used. Detection sensitivity of femtomole of both oligomer and protein has been achieved. Both positive and negative ions were observed with little fragmentation.

  15. Matrix-Free UV-Laser Desorption Ionization Mass Spectrometry as a Versatile Approach for Accelerating Dereplication Studies on Lichens.

    Science.gov (United States)

    Le Pogam, Pierre; Schinkovitz, Andreas; Legouin, Béatrice; Le Lamer, Anne-Cécile; Boustie, Joël; Richomme, Pascal

    2015-10-20

    The present study examined the suitability of laser desorption/ionization time-of-flight mass spectrometry (LDI-MS) for the rapid chemical fingerprinting of lichen extracts. Lichens are known to produce a wide array of secondary metabolites. Most of these compounds are unique to the symbiotic condition but some can be found in many species. Therefore, dereplication, that is, the rapid identification of known compounds within a complex mixture is crucial in the search for novel natural products. Over the past decade, significant advances were made in analytical techniques and profiling methods specifically adapted to crude lichen extracts, but LDI-MS has never been applied in this context. However, most classes of lichen metabolites have UV chromophores, which are quite similar to commercial matrix molecules used in matrix-assisted laser desorption ionization (MALDI). It is consequently postulated that these molecules could be directly detectable by matrix-free LDI-MS. The present study evaluated the versatility of this technique by investigating the LDI properties of a vast array of single lichen metabolites as well as lichen extracts of known chemical composition. Results from the LDI experiments were compared with those obtained by direct ESI-MS detection as well as LC-ESI-MS. It was shown that LDI ionization leads to strong molecular ion formation with little fragmentation, thus, facilitating straightforward spectra interpretation and representing a valuable alternative to time-consuming LC-MS analysis.

  16. Laser-Induced Acoustic Desorption Atmospheric Pressure Photoionization via VUV-Generating Microplasmas

    Science.gov (United States)

    Benham, Kevin; Hodyss, Robert; Fernández, Facundo M.; Orlando, Thomas M.

    2016-11-01

    We demonstrate the first application of laser-induced acoustic desorption (LIAD) and atmospheric pressure photoionization (APPI) as a mass spectrometric method for detecting low-polarity organics. This was accomplished using a Lyman-α (10.2 eV) photon generating microhollow cathode discharge (MHCD) microplasma photon source in conjunction with the addition of a gas-phase molecular dopant. This combination provided a soft desorption and a relatively soft ionization technique. Selected compounds analyzed include α-tocopherol, perylene, cholesterol, phenanthrene, phylloquinone, and squalene. Detectable surface concentrations as low as a few pmol per spot sampled were achievable using test molecules. The combination of LIAD and APPI provided a soft desorption and ionization technique that can allow detection of labile, low-polarity, structurally complex molecules over a wide mass range with minimal fragmentation.

  17. Laser-Induced Acoustic Desorption Atmospheric Pressure Photoionization via VUV-Generating Microplasmas.

    Science.gov (United States)

    Benham, Kevin; Hodyss, Robert; Fernández, Facundo M; Orlando, Thomas M

    2016-11-01

    We demonstrate the first application of laser-induced acoustic desorption (LIAD) and atmospheric pressure photoionization (APPI) as a mass spectrometric method for detecting low-polarity organics. This was accomplished using a Lyman-α (10.2 eV) photon generating microhollow cathode discharge (MHCD) microplasma photon source in conjunction with the addition of a gas-phase molecular dopant. This combination provided a soft desorption and a relatively soft ionization technique. Selected compounds analyzed include α-tocopherol, perylene, cholesterol, phenanthrene, phylloquinone, and squalene. Detectable surface concentrations as low as a few pmol per spot sampled were achievable using test molecules. The combination of LIAD and APPI provided a soft desorption and ionization technique that can allow detection of labile, low-polarity, structurally complex molecules over a wide mass range with minimal fragmentation. Graphical Abstract ᅟ.

  18. Theoretical Study of Pulsed-Laser-Induced Resonant Desorption.

    Science.gov (United States)

    1986-11-01

    laser the energy flow in the steady state is constant in time . Per unit of time it is given by,2 d- - & L Ire CW1 ( P () Pe ) " (4.4) dt r2 + 262 CW...1 - 2Pe(eq)] 2 + a cw (6.6) which is constant in time . The energy flow into the substrate is given by, dE2 cw A 2- 0 1 - 2Pe(-)1. (6.7) t F Jq, 14

  19. Detection of drugs in lifted cyanoacrylate-developed latent fingermarks using two laser desorption/ionisation mass spectrometric methods.

    Science.gov (United States)

    Sundar, Latha; Rowell, Frederick

    2014-02-01

    This paper describes a method for lifting cyanoacrylate (CNA)-developed latent fingermarks from a glass surface and the detection of five drugs in lifted marks from fingers that had been in contact with the drugs, using Surface Assisted Laser Desorption Ionisation Time of Flight Mass Spectrometry (SALDI-TOF-MS) or Matrix Assisted Laser Desorption Ionisation TOF-MS (MALDI-TOF-MS). Two drugs of abuse (cocaine and methadone) and three therapeutic drugs (aspirin, paracetamol and caffeine) were used as contact residues. Latent fingermarks spiked with the drugs were subjected to CNA fuming followed by dusting with ARRO SupraNano™ MS black magnetic powder (SALDI-TOF-MS) or 2,5-dihydroxybenzoic acid (DHB) (MALDI-TOF-MS). The dusted mark was then exposed to solvent vapour before lifting with a commercial fingerprint lifting tape following established procedures. The presence of the drugs was then confirmed by direct analysis on the tape without further processing using SALDI- or MALDI-TOF-MS. The black magnetic fingerprint powder provided visual enhancement of the CNA-fingermark while no visual enhancement was observed for marks dusted with DHB powder. Similar [M + H](+) peaks for all the drug analytes were observed for both methods along with some sodium and potassium adducts for SALDI-MS and some major fragment ions but the SALDI signals were generally more intense. Simple exposure to acetone vapour of the CNA-developed marks enabled their effective transfer onto the tape which was crucial for subsequent MS detection of the analytes.

  20. Brominated Tyrosine and Polyelectrolyte Multilayer Analysis by Laser Desorption VUV Postionization and Secondary Ion Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    University of Illinois at Chicago; Blaze, Melvin M. T.; Takahashi, Lynelle; Zhou, Jia; Ahmed, Musahid; Gasper, Gerald; Pleticha, F. Douglas; Hanley, Luke

    2011-03-14

    The small molecular analyte 3,5-dibromotyrosine (Br2Y) and chitosan-alginate polyelectrolyte multilayers (PEM) with and without adsorbed Br2Y were analyzed by laser desorption postionization mass spectrometry (LDPI-MS). LDPI-MS using 7.87 eV laser and tunable 8 ? 12.5 eV synchrotron vacuum ultraviolet (VUV) radiation found that desorption of clusters from Br2Y films allowed detection by≤8 eV single photon ionization. Thermal desorption and electronic structure calculations determined the ionization energy of Br2Y to be ~;;8.3?0.1 eV and further indicated that the lower ionization energies of clusters permitted their detection at≤8 eV photon energies. However, single photon ionization could only detect Br2Y adsorbed within PEMs when using either higher photon energies or matrix addition to the sample. All samples were also analyzed by 25 keV Bi3 + secondary ion mass spectrometry (SIMS), with the negative ion spectra showing strong parent ion signal which complemented that observed by LDPI-MS. The negative ion SIMS depended strongly on the high electron affinity of this specific analyte and the analyte?s condensed phase environment.

  1. Laser-assisted serial tonsillectomy.

    Science.gov (United States)

    Krespi, Y P; Ling, E H

    1994-10-01

    Laser ablation of the palatine tonsils is a useful alternative to tonsillectomy in adults. Cryptic tonsillitis is a common problem causing recurrent infection, sore throat, and halitosis. Elimination and/or obliteration of surface pockets (crypts) of the palatine tonsils utilizing the CO2 laser was effective in 86 patients treated in the past 4 years. Ablation of the tonsil surface was performed in stages under local anesthesia in an office setting; CO2 laser energy delivered through the "SwiftLase" handpiece extension provided char-free, superficial layer ablation of tissue. "SwiftLase" is easily installed onto existing CO2 laser units and provides high-power densities by utilizing a focused laser beam in an extremely fast uniform scan over an extended area (up to 4 mm) within a fraction of a second. This method and results of its use are discussed.

  2. Measurement of desorption energies of H 2 and CO from SS-304 LN and inconel 625 surfaces by laser induced thermal desorption

    Science.gov (United States)

    Tagle, J. A.; Pospieszczyk, A.

    Stainless steel (AISI 304 LN) and Inconel 625 surfaces have been exposed to different pressures of H 2 and CO in the temperature range of 300-473 K. A laser heating technique for studying fast surface processes was applied to determine the desorption energies of these gasses from SS 304 LN and Inconel 625 surfaces. The Clausius-Clapeyron plots give a desorption energy value of 97 kJ mol -1 for H 2/SS 304 LN and 77.4 kJ mol -1 for H 2/Inconel 625. The adsorption of CO on these surfaces is drastically affected by the fast growing of a chromium oxide layer at the surface during the laser shots. In this case the desorption energy was determined by fitting both the experimental recoverage times and the equilibrium coverage curves. The oxide passivation layer produces a decrease of the desorption energy from 92.8 to 72.8 kJ mol -1 for CO/SS 304 LN and from 91.8 to 77.9 kJ mol -1 for CO/Inconel 625 when the oxygen surface concentration increases to 14%.

  3. Laser Desorption Postionization Mass Spectrometry of Antibiotic-Treated Bacterial Biofilms using Tunable Vacuum Ultraviolet Radiation

    Energy Technology Data Exchange (ETDEWEB)

    Gasper, Gerald L; Takahashi, Lynelle K; Zhou, Jia; Ahmed, Musahid; Moore, Jerry F; Hanley, Luke

    2010-08-04

    Laser desorption postionization mass spectrometry (LDPI-MS) with 8.0 ? 12.5 eV vacuum ultraviolet synchrotron radiation is used to single photon ionize antibiotics andextracellular neutrals that are laser desorbed both neat and from intact bacterial biofilms. Neat antibiotics are optimally detected using 10.5 eV LDPI-MS, but can be ionized using 8.0 eV radiation, in agreement with prior work using 7.87 eV LDPI-MS. Tunable vacuum ultraviolet radiation also postionizes laser desorbed neutrals of antibiotics and extracellular material from within intact bacterial biofilms. Different extracellular material is observed by LDPI-MS in response to rifampicin or trimethoprim antibiotic treatment. Once again, 10.5 eV LDPI-MS displays the optimum trade-off between improved sensitivity and minimum fragmentation. Higher energy photons at 12.5 eV produce significant parent ion signal, but fragment intensity and other low mass ions are also enhanced. No matrix is added to enhance desorption, which is performed at peak power densities insufficient to directly produce ions, thus allowing observation of true VUV postionization mass spectra of antibiotic treated biofilms.

  4. Processing of C60 thin films by Matrix-Assisted Pulsed Laser Evaporation (MAPLE)

    DEFF Research Database (Denmark)

    Canulescu, Stela; Schou, Jørgen; Fæster, Søren

    2011-01-01

    has been applied for deposition of fullerenes for the first time and we have studied the growth of thin films of solid C60. The fragmentation of C60 fullerene molecules induced by ns ablation in vacuum of a frozen anisole target with C60 was investigated by matrix-assisted laser desorption......Thin films of fullerenes (C60) were deposited onto silicon using matrix-assisted pulsed laser evaporation (MAPLE). The deposition was carried out from a frozen homogeneous dilute solution of C60 in anisole (0.67 wt%), and over a broad range of laser fluences, from 0.15 J/cm2 up to 3.9 J/cm2. MAPLE....../ionization (MALDI). Our findings show that intact fullerene films can be produced with laser fluences ranging from 0.15 J/cm2 up to 1.5 J/cm2....

  5. Laser assisted foaming of aluminum

    Energy Technology Data Exchange (ETDEWEB)

    Kathuria, Y.P. [Laser X Co. Ltd., Aichi (Japan)

    2001-09-01

    Recently aluminum foams have evoked considerable interest as an alternative material owing to their wide range of applications ranging from microelectronics, through automobiles to aerospace industries. The manufacturing techniques and characterization methods for aluminum foams require further development to achieve effective and economical use of this material. In this communication the authors demonstrate the feasibility of unidirectional and localized expansion of the aluminum foam using the Nd-YAG/CO{sub 2} laser and powder metallurgy. (orig.)

  6. Applications of electrospray laser desorption ionization mass spectrometry for document examination.

    Science.gov (United States)

    Cheng, Sy-Chyi; Lin, Yu-Shan; Huang, Ming-Zong; Shiea, Jentaie

    2010-01-01

    We have employed electrospray laser desorption ionization mass spectrometry (ELDI-MS) to rapidly characterize certain classes of compounds--the inks within the characters made by inks and inkjet printer on regular paper and the chemical compounds within thermal papers. This ELDI-MS approach allowed the ink and paper samples to be distinguished in terms of their chemical compositions. Sample pretreatment was unnecessary and the documents were practically undamaged after examination. The ink chemicals on the documents were desorbed through laser irradiation (sampling spot area: <100 microm(2)); the desorbed molecules then entered an electrospray plume--prepared from an acidic methanol/water solution (50%)--where they became ionized through fusion or ion-molecule reactions with the charged solvent species and droplets in the plume.

  7. 爆炸条件下溶菌酶反应产物的基质辅助激光解吸-飞行时间质谱分析%Analysis of Reaction Products of Lysozyme under the Explosion Condition by Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    刘素红; 夏攀; 张成功; 张立; 郭寅龙

    2014-01-01

    Identification and determination of explosives and explosive residues were a subject of continuing strong interest in analytical chemistry and forensic science. In this paper, the reaction products of lysozyme under the explosion condition were analyzed by a MALDI-TOFMS (Matrix-assisted laser desorption ionization time-of-flight mass spectrometry) method. There was no difference in the tryptic digest between the normal lysozyme and the reaction products generated by detonator, while some adduct peaks such as [MH+17]+, [MH+18]+, [MH+28]+, [MH+32]+, and [MH+44]+ were discovered in the explosives. This may be attributed to the reaction between the lysozyme and the active small molecule gases such as NH3, H2O, CO/N2, O2, CO2, which were generated during the explosion. Characteristic peaks which were produced by lysozyme and the active small molecule gases from different explosives can be used to discriminate the six explosives. For example, H2O molecules which were generated during the exploration by tri-nitrotoluene (TNT) can specifically react with VFGRCE-LAAAMKRHGLDNYR (m/z 2307) to produce a characteristic peak at m/z 2325 (2307+18). Also, H2O molecules which were generated by hexahydro-1,3,5-trinitroazine (RDX) can completely react with IVSDGNGMNAWVAWRNRCK (m/z 2177) to produce a characteristic peak at m/z (2177+18). Characteristic peak at m/z 1301 was produced by GYSLGNWVCAAK (m/z 1269) and O2 molecules for the identification of pentaerythritol tetranitrate (PETN). While for black powder, O2 and H2O can both react with IVSDGNGMNAWVAWR (m/z 1676) to produce product ions peaks at m/z 1694 and 1708. However, only the O2 molecules can react with IVSDGNGMNAWVAWR for pyrotechnic composition. As for ammon explosive, which is a mixture of inorganic explosives and organic explosives, CO2 molecules can react with a plurality of reac-tion sites of lysozyme to produce a series of characteristic peaks signals such as m/z 1313 (1269+44), 1720 (1676+44), 1848 (1804+44), 2722

  8. 基质辅助激光解析/电离飞行时间质谱仪检测KPC型碳青霉烯酶的研究%Detection of KPC carbapenem by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    胡燕燕; 孙谦; 蔡加昌; 周宏伟; 陈功祥; 张嵘

    2012-01-01

    目的 用基质辅助激光解析/电离飞行时间质谱仪(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF MS)比较产KPC型碳青霉烯酶肠杆菌科细菌在不同药物浓度下水解厄他培南的能力.方法 收集浙江大学医学院附属第二医院临床分离的19株单产KPC肠杆菌科细菌,包括10株奇异变形杆菌,3株产气肠杆菌,2株黏质沙雷菌,2株弗劳地柠檬酸杆菌,1株肺炎克雷伯菌,1株阴沟肠杆菌;杭州市中医院分离的7株产KPC摩根摩根菌,以及上述7株摩根摩根菌和部分奇异变形杆菌(4株)的大肠埃希菌转移接合子.用MALDI-TOF MS检测产KPC肠杆菌科细菌水解厄他培南的能力.结果 当厄他培南浓度为0.1 g/L时,37株单产KPC型碳青霉烯酶的肠杆菌科细菌在1.5h内全部水解厄他培南,质谱图显示厄他培南所呈现的3个峰全部消失,灵敏度高达100%;当厄他培南浓度为0.3 g/L时,1.5h组水解厄他培南的灵敏度为70.3% (26/37),2.5h组为89.2% (33/37),3.5h组为94.6% (35/37);当厄他培南浓度为0.5 g/L时,1.5h组水解厄他培南的灵敏度为48.6% (18/37),2.5h组为83.8% (31/37),3.5h组为94.6%( 35/37).两独立样本非参数检验统计显示:厄他培南0.1 g/L组与0.3 g/L、0.5 g/L组之间的水解时间差异有统计学意义,而0.3 g/L与0.5 g/L组之间的水解时间差异无统计学意义;细菌组内数据统计显示,除奇异变形杆菌组与大肠埃希菌组之间差异有统计学意义外,其他各细菌组之间差异没有统计学意义.结论 MALDI-TOF MS操作简便,可快速检测产KPC型碳青霉烯酶的肠杆菌科细菌,敏感率高,假阳性率低,适合微生物检验中用于产KPC型碳青霉烯酶的检测,推荐使用0.1 g/L的厄他培南作为水解底物来检测产KPC型肠杆菌科细菌.%Objective To compare the capability of ertapenem-hydrolyzing in various concentrations in KPC-producing Enterobacteriaceae by matrix-assisted

  9. Mechanically assisted laser forming of thin beams

    Science.gov (United States)

    Mucha, Zygmunt; Widłaszewski, Jacek; Kurp, Piotr; Mulczyk, Krystian

    2016-12-01

    Laser-assisted forming techniques have been developed in recent years to aid plastic working of materials, which are difficult in processing at normal temperatures due to a high brittleness, effects of high work-hardening or a high spring-back phenomenon. This paper reports initial experimental investigations and numerical simulations of a mechanically-assisted laser forming process. The research is aimed at facilitating plastic shaping of thin-walled parts made of high temperature resistant alloys. Stainless steel plate, 1 mm thick, 20 mm wide, was mounted in the cantilever arrangement and a gravitational load was applied to its free end. A CO2 laser beam with rectangular cross-section traversed along the plate, towards the fixed edge. Laser spot covered the whole width of the plate. Experiments and simulations using the finite element method were performed for different values of mechanical load and with constant laser processing parameters. Experimentally validated numerical model allowed analysis of plastic deformation mechanism under the hybrid thermo-mechanical processing. The revealed mechanism of deformation consists in intense material plastic flow near the laser heated surface. This behavior results mainly from the tension state close to the heated surface and the decrease of material yield stress at elevated temperature. Stress state near the side edges of the processed plate favored more intense plastic deformation and the involved residual stress in this region.

  10. Laser assisted Drug Delivery: Grundlagen und Praxis.

    Science.gov (United States)

    Braun, Stephan Alexander; Schrumpf, Holger; Buhren, Bettina Alexandra; Homey, Bernhard; Gerber, Peter Arne

    2016-05-01

    Die topische Applikation von Wirkstoffen ist eine zentrale Therapieoption der Dermatologie. Allerdings mindert die effektive Barrierefunktion der Haut die Bioverfügbarkeit der meisten Externa. Fraktionierte ablative Laser stellen ein innovatives Verfahren dar, um die epidermale Barriere standardisiert, kontaktfrei zu überwinden. Die Bioverfügbarkeit im Anschluss applizierter Externa wird im Sinne einer laser assisted drug delivery (LADD) signifikant gesteigert. Das Prinzip der LADD wird bereits in einigen Bereichen der Dermatologie erfolgreich eingesetzt. Die vorliegende Übersichtsarbeit soll einen Überblick über die aktuellen aber auch perspektivischen Einsatzmöglichkeiten der LADD bieten.

  11. Laser-assisted photoelectric effect from surfaces.

    Science.gov (United States)

    Miaja-Avila, L; Lei, C; Aeschlimann, M; Gland, J L; Murnane, M M; Kapteyn, H C; Saathoff, G

    2006-09-15

    We report the first observation of the laser-assisted photoelectric effect from a solid surface. By illuminating a Pt(111) sample simultaneously with ultrashort 1.6 eV and 42 eV pulses, we observe sidebands in the extreme ultraviolet photoemission spectrum. The magnitude of these sidebands as a function of time delay between the laser and extreme ultraviolet pulses represents a cross-correlation measurement of the extreme ultraviolet pulse. This effect promises to be useful to extend extreme ultraviolet pulse duration measurements to higher photon energies, as well as opening up femtosecond-to-attosecond time-scale electron dynamics in solid and surface-adsorbate systems.

  12. Laser desorption time-of-flight mass spectrometry of vacuum UV photo-processed methanol ice

    Science.gov (United States)

    Paardekooper, D. M.; Bossa, J.-B.; Linnartz, H.

    2016-07-01

    Context. Methanol in the interstellar medium mainly forms upon sequential hydrogenation of solid CO. With typical abundances of up to 15% (with respect to water) it is an important constituent of interstellar ices where it is considered as a precursor in the formation of large and complex organic molecules (COMs), e.g. upon vacuum UV (VUV) photo-processing or exposure to cosmic rays. Aims: This study aims at detecting novel complex organic molecules formed during the VUV photo-processing of methanol ice in the laboratory using a technique more sensitive than regular surface diagnostic tools. In addition, the formation kinetics of the main photo-products of methanol are unravelled for an astronomically relevant temperature (20 K) and radiation dose. Methods: The VUV photo-processing of CH3OH ice is studied by applying laser desorption post-ionisation time-of-flight mass spectrometry (LDPI TOF-MS), and analysed by combining molecule-specific fragmentation and desorption features. Results: The mass spectra correspond to fragment ions originating from a number of previously recorded molecules and from new COMs, such as the series (CO)xH, with x = 3 and y prebiotic glycerin belongs. The formation of these large COMs has not been reported in earlier photolysis studies and suggests that such complex species may form in the solid state under interstellar conditions.

  13. Fractional laser-assisted topical delivery leads to enhanced, accelerated and deeper cutaneous 5-fluorouracil uptake

    DEFF Research Database (Denmark)

    Wenande, Emily; Olesen, Uffe H; Nielsen, Mette M B

    2016-01-01

    and biodistribution. METHODS: In vitro porcine skin underwent AFXL-exposure using a fractional 10,600nm CO2-laser, generating microscopic ablation zones (MAZ) reaching the dermoepidermal junction (MAZ-ED), superficial-(MAZ-DS), or mid-dermis(MAZ-DM). 5-FU in AFXL-exposed and control skin was measured in Franz......BACKGROUND: Topical 5-Fluorouracil (5-FU) exhibits suboptimal efficacy for non-melanoma skin cancer, attributed to insufficient intracutaneous penetration. This study investigates the impact of ablative fractional laser (AFXL) at different laser-channel depths on cutaneous 5-FU pharmacokinetics...... diffusion cells at 4 and 24 hours (n =55). HPLC quantified 5-FU in full-thickness skin, specific skin depths of 100μm-1500μm, and transcutaneous receiver-compartments. Qualitative matrix-assisted laser-desorption/ionization mass-spectrometry imaging (MALDI-MSI) visualized 5-FU in selected samples. RESULTS...

  14. Atmospheric pressure laser desorption/ionization using a 6-7 µm-band mid-infrared tunable laser and liquid water matrix.

    Science.gov (United States)

    Hiraguchi, Ryuji; Hazama, Hisanao; Masuda, Katsuyoshi; Awazu, Kunio

    2015-01-01

    Due to the characteristic absorption peaks in the IR region, various molecules can be used as a matrix for infrared matrix-assisted laser desorption/ionization (IR-MALDI). Especially in the 6-7 µm-band IR region, solvents used as the mobile phase for liquid chromatography have absorption peaks that correspond to their functional groups, such as O-H, C=O, and CH3. Additionally, atmospheric pressure (AP) IR-MALDI, which is applicable to liquid-state samples, is a promising technique to directly analyze untreated samples. Herein we perform AP-IR-MALDI mass spectrometry of a peptide, angiotensin II, using a mid-IR tunable laser with a tunable wavelength range of 5.50-10.00 µm and several different matrices. The wavelength dependences of the ion signal intensity of [M + H](+) of the peptide are measured using a conventional solid matrix, α-cyano-4-hydroxycinnamic acid (CHCA) and a liquid matrix composed of CHCA and 3-aminoquinoline. Other than the O-H stretching and bending vibration modes, the characteristic absorption peaks are useful for AP-IR-MALDI. Peptide ions are also observed from an aqueous solution of the peptide without an additional matrix, and the highest peak intensity of [M + H](+) is at 6.00 µm, which is somewhat shorter than the absorption peak wavelength of liquid water corresponding to the O-H bending vibration mode. Moreover, long-lasting and stable ion signals are obtained from the aqueous solution. AP-IR-MALDI using a 6-7 µm-band IR tunable laser and solvents as the matrix may provide a novel on-line interface between liquid chromatography and mass spectrometry.

  15. Identification of colorants in pigmented pen inks by laser desorption mass spectrometry.

    Science.gov (United States)

    Papson, Kaitlin; Stachura, Sylwia; Boralsky, Luke; Allison, John

    2008-01-01

    Pigments are rapidly replacing dyes as colorants in pen and printer inks, due to their superior colors and stability. Unfortunately, tools commonly used in questioned document examination for analyzing pen inks, such as TLC, cannot be used for the analysis of insoluble pigments on paper. Laser desorption mass spectrometry is demonstrated here as a tool for analyzing pigment-based pen inks. A pulsed nitrogen laser can be focused onto a pen stroke from a pigmented ink pen on paper, and positive and negative ions representative of the pigment can be generated for subsequent mass spectrometric analysis. Targeted pens for this work were a set of Uni-ball 207 pigmented ink pens containing blue, light blue, orange, green, violet, red, pink, and black inks. Copper phthalocyanine was identified as the pigment used to make both blue inks. A mixture of halogenated copper phthalocyanines were identified in the green ink. Unexpectedly, the pink ink was found to contain a red pigment, Pigment Red 12, treated with a mixture of water-soluble dyes. Each sample yielded ions representative of the pigments present.

  16. Quantum dynamics of STM and laser induced desorption of atoms and molecules from surfaces

    CERN Document Server

    Boendgen, G

    2001-01-01

    The manipulation of atoms and molecules at solid surfaces by electronic excitations with electrons (or holes) emitted from the tip of a scanning tunneling microscope (STM) or with laser radiation is both of applied and fundamental interest, e.g. for micro- and nanostructuring of materials, the clarification of elementary (catalytic) reaction mechanisms and for the question of how to treat the quantum dynamics of a laser or STM driven 'system' (the adsorbate) in contact with a dissipative (energy-withdrawing) 'bath' (the substrate). Desorption induced by electronic transitions (DIET) and its variant DIMET (M = multiple) are among the simplest possible 'reactions' of adsorbate-surface systems; usually involving extremely short-lived electronically excited intermediates. In this thesis, the ultra-short dynamics of directly (localised to the adsorbate-substrate complex) and indirectly (i.e., through the substrate) stimulated DIET and DIMET processes was studied for Si(100)-(2x1):H(D) and Pt(111):NO. Isotope effec...

  17. Laser-Assisted Photoelectric Effect from Liquids

    Science.gov (United States)

    Arrell, C. A.; Ojeda, J.; Mewes, L.; Grilj, J.; Frassetto, F.; Poletto, L.; van Mourik, F.; Chergui, M.

    2016-09-01

    The laser-assisted photoelectric effect from liquid surfaces is reported for the first time. Photoelectrons generated by 35.6 eV radiation from a liquid microjet of water under vacuum are dressed with a ℏω =1.55 eV laser field. The subsequent redistribution of the photoelectron energies consists in the appearance of sidebands shifted by energies equivalent to ℏω , 2 ℏω , and 3 ℏω . The response has been modeled to the third order and combined with energy-resolved measurements. This result opens the possibility to investigate the dynamics at surfaces of liquid solutions and provide information about the electron emission process from a liquid.

  18. Quantum Electrodynamics Theory of Laser Assisted Recombination

    Institute of Scientific and Technical Information of China (English)

    敖淑艳; 程太旺; 李晓峰; 潘守甫; 傅盘铭

    2003-01-01

    Using a formal scattering theoretical approach, we develop a nonperturbative quantum electrodynamics theory to describe laser assisted recombination (LAR), in which an electron initially in the quantized Volkov state recombines with an ion and emits a high-energy photon with frequency defined by energy conservation laws.The transition probability is expressed as an analytic closed form and the spectrum of LAR reflects mainly the properties of general Bessel functions. For the case of a fast electron the LAR spectrum is confined in a well-defined range, while for a slow electron, the LAR spectrum exhibits a double-plateau structure.

  19. Laser-assisted delivery of topical methotrexate - in vitro investigations.

    Science.gov (United States)

    Taudorf, Elisabeth Hjardem

    2016-06-01

    Ablative fractional lasers (AFXL) are increasingly used to treat dermatological disorders and to facilitate laser-assisted topical drug delivery. In this thesis, laser-tissue interactions generated by stacked pulses with a miniaturized low-power 2,940 nm AFXL were characterized (study I). Knowledge of the correlation between laser parameters and tissue effects was used to deliver methotrexate (MTX) topically through microscopic ablation zones (MAZs) of precise dimensions. MTX is a well-known chemotherapeutic and anti-inflammatory drug that may cause systemic adverse effects, and topical delivery is thus of potential benefit. The impact of MAZ depth (study II) and transport kinetics (study III) on MTX deposition in skin as well as transdermal permeation was determined in vitro. Quantitative analyses of dermal and transdermal MTX concentrations were performed by high performance liquid chromatography (HPLC) (study II & III), while qualitative analyses of MTX biodistribution in skin were illustrated and semi-quantified by fluorescence microscopy (study II & III) and desorption electro spray mass spectrometry imaging (DESI-MSI) (study III). Laser-tissue interactions generated by AFXL: AFXL-exposure generated a variety of MAZ-dimensions. MAZ depth increased linearly with the logarithm of total energy delivered by stacked pulses, but was also affected by variations in power, pulse energy, pulse duration, and pulse repetition rate. Coagulation zones lining MAZs increased linearly with the applied total energy, while MAZ width increased linearly with the logarithm of stacked pulses. Results were gathered in a mathematical model estimating relations between laser parameters and specific MAZ dimensions. Impact of MAZ depth on AFXL-assisted topical MTX delivery: Pretreatment by AFXL facilitated topical MTX delivery to all skin layers. Deeper MAZs increased total MTX deposition in skin compared to superficial MAZs and altered the intradermal biodistribution profile towards

  20. Charging-assisted desorption of deuterium films by keV electrons

    DEFF Research Database (Denmark)

    Schou, Jørgen; Thestrup Nielsen, Birgitte; Pedersen, Thomas Garm

    2009-01-01

    m. The initial film thickness and the mass loss as result of desorption were monitored by the QCM. The electron beam current was kept at about or below 100 nA to avoid beam-induced evaporation. Secondary electron emission was suppressed to a value below 0.01-0.03 electrons/electron by a repeller...

  1. DIFFERENTIATION OF AEROMONAS ISOLATES OBTAINED FROM DRINKING WATER DISTRIBUTION SYSTEM USING MATRIX-ASSISTED LASER DESCRIPTION/IONIZATION-MASS SPECTROMETRY (MALDI-MS)

    Science.gov (United States)

    The genus Aeromonas is one of several medically significant genera that have gained prominence due to their evolving taxonomy and controversial role in human diseases. In this study, matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was used to analyze the...

  2. UV–UV hole burning and IR dip spectroscopy of homophenylalanine by laser desorption supersonic jet technique

    Energy Technology Data Exchange (ETDEWEB)

    Sohn, Woon Yong; Ishiuchi, Shun-ichi [Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8503 (Japan); Çarçabal, Pierre [Institut des Sciences Moléculaires d’Orsay, CNRS, Université Paris Sud XI, Bâtiment 210, 91405 Orsay (France); Oba, Hikari [Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8503 (Japan); Fujii, Masaaki, E-mail: mfujii@res.titech.ac.jp [Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8503 (Japan)

    2014-12-05

    Highlights: • Homophenylalanine was measured by laser desorption supersonic jet spectroscopy. • Ten conformers were found by UV–UV hole burning spectroscopy. • The observed conformers were assigned by the combination of UV and IR spectra. • Franck–Condon simulations were employed to assign the flexible molecule. - Abstract: Conformer selected electronic and vibrational spectra of homophenylalanine, phenylalanine analogue molecule, were measured by UV–UV hole burning and IR dip spectroscopy combined with laser desorption technique. 10 conformers were found by UV–UV hole burning spectroscopy and their structures were assigned by IR dip and UV absorption spectra with aid of quantum chemical calculations in both S{sub 0} and S{sub 1}. This study shows that the combination of simulated IR and UV spectra is powerful to assign flexible molecules.

  3. TiO(2) printed aluminum foil: single-use film for a laser desorption/ionization target plate.

    Science.gov (United States)

    Bi, Hongyan; Qiao, Liang; Busnel, Jean-Marc; Devaud, Valerie; Liu, Baohong; Girault, Hubert H

    2009-02-01

    Single-use aluminum foil-based laser desorption/ionization (LDI) target plates have been developed for mass spectrometry (MS) analysis and provide detection results comparable to those of commercial stainless steel plates while offering a convenient way to avoid the time-consuming surface cleaning process. Additionally, arrays of TiO(2) nanoparticle spots are coated on the foil either by screen-printing or rotogravure-printing followed by sintering to form a mesoporous layer spot to act as an anchor for sample deposition. These TiO(2) spots offer further functions to the Al foil, such as matrix-free laser desorption/ionization or specific affinity for in situ enrichment of phosphopeptides. The single-use TiO(2)-Al foils are cheap to produce, easy to use, and well suited for high-throughput proteomics research. They can also be of interest for protein post-translational modifications study.

  4. Photoionization and Dissociative Photoionization Study of Cholesterol by IR Laser Desorption/Tunable Synchrotron VUV Photoionization Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    Yang Pan; Li-dong Zhang; Hui-jun Guo; Hao Yin; Fei Qi

    2009-01-01

    Elementary cholesterol was analyzed with IR laser desorption/tunable synchrotron vacuum ultraviolet photoionization mass spectrometry. An exclusive molecular ion of cholesterol is observed by near threshold single-photon ionization with high efficiency. Fragments are yielded with the increase of photon energy. The structures of various fragments are determined with commercial electron ionization time-of-flight mass spectrometry. Dominant fragmentation pathways are discussed in detail with the aid of ab initio calculations.

  5. Structural features of lipoarabinomannan from Mycobacterium bovis BCG. Determination of molecular mass by laser desorption mass spectrometry.

    Science.gov (United States)

    Venisse, A; Berjeaud, J M; Chaurand, P; Gilleron, M; Puzo, G

    1993-06-15

    It was recently shown that mycobacterial lipoarabinomannan (LAM) can be classified into two types (Chatterjee, D., Lowell, K., Rivoire B., McNeil M. R., and Brennan, P. J. (1992) J. Biol. Chem. 267, 6234-6239) according to the presence or absence of mannosyl residues (Manp) located at the nonreducing end of the oligoarabinosyl side chains. These two types of LAM were found in a pathogenic Mycobacterium tuberculosis strain and in an avirulent M. tuberculosis strain, respectively, suggesting that LAM with Manp characterizes virulent and "disease-inducing strains." We now report the structure of the LAM from Mycobacterium bovis Bacille Calmette-Guérin (BCG) strain Pasteur, largely used throughout the world as vaccine against tuberculosis. Using an up-to-date analytical approach, we found that the LAM of M. bovis BCG belongs to the class of LAMs capped with Manp. By means of two-dimensional homonuclear and heteronuclear scalar coupling NMR analysis and methylation data, the sugar spin system assignments were partially established, revealing that the LAM contained two types of terminal Manp and 2-O-linked Manp. From the following four-step process: (i) partial hydrolysis of deacylated LAM (dLAM), (ii) oligosaccharide derivatization with aminobenzoic ethyl ester, (iii) HPLC purification, (iv) FAB/MS-MS analysis; it was shown that the dimannosyl unit alpha-D-Manp-(1-->2)-alpha-D-Manp is the major residue capping the termini of the arabinan of the LAM. In this report, LAM molecular mass determination was established using matrix-assisted UV-laser desorption/ionization mass spectrometry which reveals that the LAM molecular mass is around 17.4 kDa. The similarity of the LAM structures between M. bovis BCG and M. tuberculosis H37Rv is discussed in regard to their function in the immunopathology of mycobacterial infection.

  6. Inkjet-printed gold nanoparticle surfaces for the detection of low molecular weight biomolecules by laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Marsico, Alyssa L M; Creran, Brian; Duncan, Bradley; Elci, S Gokhan; Jiang, Ying; Onasch, Timothy B; Wormhoudt, Joda; Rotello, Vincent M; Vachet, Richard W

    2015-11-01

    Effective detection of low molecular weight compounds in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is often hindered by matrix interferences in the low m/z region of the mass spectrum. Here, we show that monolayer-protected gold nanoparticles (AuNPs) can serve as alternate matrices for the very sensitive detection of low molecular weight compounds such as amino acids. Amino acids can be detected at low fmol levels with minimal interferences by properly choosing the AuNP deposition method, density, size, and monolayer surface chemistry. By inkjet-printing AuNPs at various densities, we find that AuNP clusters are essential for obtaining the greatest sensitivity. Graphical Abstract ᅟ.

  7. Laser-assisted growth of carbon nanotubes - A review

    NARCIS (Netherlands)

    Burgt, Y. van de

    2014-01-01

    Laser-assisted chemical vapor deposition (LACVD) is an attractive maskless process for growing locally carbon nanotubes at selected places on substrates that may contain temperature-sensitive components. This review gives a comprehensive overview of the reported research with respect to laser assist

  8. Developments and Applications of Electrophoresis and Small Molecule Laser Desorption Ionization Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Hui [Iowa State Univ., Ames, IA (United States)

    2007-01-01

    Ultra-sensitive native fluorescence detection of proteins with miniaturized one- and two-dimensional polyacrylamide gel electrophoresis was achieved with laser side-entry excitation, which provides both high excitation power and low background level. The detection limit for R-phycoerythrin protein spots in 1-D SDS-PAGE was as low as 15 fg, which corresponds to 40 thousand molecules only. The average detection limit of six standard native proteins was 5 pg per band and the dynamic range spanned more than 3 orders of magnitude. Approximately 150 protein spots from 30 ng of total Escherichia coli extraction were detected on a 0.8 cm x 1 cm gel in two-dimensional separation. Estrogen-DNA adducts as 4-OHE1(E2)-1-N3Ade and 4-OHEI(E2)-2-NacCys were hypothesized as early risk assessment of prostate and breast cancers. Capillary electrophoresis, luminescence/absorption spectroscopy and LC-MS were used to characterize and detect these adducts. Monoclonal antibodies against each individual adduct were developed and used to enrich such compounds from urine samples of prostate and breast cancer patients as well as healthy people. Adduct 4-OHE1-1-N3Ade was detected at much higher level in urine from subjects with prostate cancer patients compared to healthy males. The same adduct and 4-OHEI-2-NacCys were also detected at a much higher level in urine from a woman with breast carcinoma than samples from healthy controls. These two DNA adducts may serve as novel biomarkers for early diagnostic of cancers. The adsorption properties of R-phycoerythrin (RPE), on the fused-silica surface were studied using capillary electrophoresis (CE) and single molecule spectroscopy. The band shapes and migration times were measured in CE. Adsorption and desorption events were recorded at the single-molecule level by imaging of the evanescent-field layer using total internal reflection. The adsorbed RPE molecules on the fused-silica prism surface were

  9. Fractional laser-assisted delivery of methyl aminolevulinate

    DEFF Research Database (Denmark)

    Haak, Christina S; Farinelli, William A; Tam, Joshua;

    2012-01-01

    Pretreatment of skin with ablative fractional lasers (AFXL) enhances the uptake of topical photosensitizers used in photodynamic therapy (PDT). Distribution of photosensitizer into skin layers may depend on depth of laser channels and incubation time. This study evaluates whether depth...... of intradermal laser channels and incubation time may affect AFXL-assisted delivery of methyl aminolevulinate (MAL)....

  10. Fuzzy model for Laser Assisted Bending Process

    Directory of Open Access Journals (Sweden)

    Giannini Oliviero

    2016-01-01

    Full Text Available In the present study, a fuzzy model was developed to predict the residual bending in a conventional metal bending process assisted by a high power diode laser. The study was focused on AA6082T6 aluminium thin sheets. In most dynamic sheet metal forming operations, the highly nonlinear deformation processes cause large amounts of elastic strain energy stored in the formed material. The novel hybrid forming process was thus aimed at inducing the local heating of the mechanically bent workpiece in order to decrease or eliminate the related springback phenomena. In particular, the influence on the extent of springback phenomena of laser process parameters such as source power, scan speed and starting elastic deformation of mechanically bent sheets, was experimentally assessed. Consistent trends in experimental response according to operational parameters were found. Accordingly, 3D process maps of the extent of the springback phenomena according to operational parameters were constructed. The effect of the inherent uncertainties on the predicted residual bending caused by the approximation in the model parameters was evaluated. In particular, a fuzzy-logic based approach was used to describe the model uncertainties and the transformation method was applied to propagate their effect on the residual bending.

  11. Cell patterning by laser-assisted bioprinting.

    Science.gov (United States)

    Devillard, Raphaël; Pagès, Emeline; Correa, Manuela Medina; Kériquel, Virginie; Rémy, Murielle; Kalisky, Jérôme; Ali, Muhammad; Guillotin, Bertrand; Guillemot, Fabien

    2014-01-01

    The aim of tissue engineering is to produce functional three-dimensional (3D) tissue substitutes. Regarding native organ and tissue complexity, cell density and cell spatial 3D organization, which influence cell behavior and fate, are key parameters in tissue engineering. Laser-Assisted Bioprinting (LAB) allows one to print cells and liquid materials with a cell- or picoliter-level resolution. Thus, LAB seems to be an emerging and promising technology to fabricate tissue-like structures that have the physiological functionality of their native counterparts. This technology has additional advantages such as automation, reproducibility, and high throughput. It makes LAB compatible with the (industrial) fabrication of 3D constructs of physiologically relevant sizes. Here we present exhaustively the numerous steps that allow printing of viable cells with a well-preserved micrometer pattern. To facilitate the understanding of the whole cell patterning experiment using LAB, it is discussed in two parts: (1) preprocessing: laser set-up, bio-ink cartridge and bio-paper preparation, and pattern design; and (2) processing: bio-ink printing on the bio-paper.

  12. CO2 adsorption-assisted CH4 desorption on carbon models of coal surface: A DFT study

    Science.gov (United States)

    Xu, He; Chu, Wei; Huang, Xia; Sun, Wenjing; Jiang, Chengfa; Liu, Zhongqing

    2016-07-01

    Injection of CO2 into coal is known to improve the yields of coal-bed methane gas. However, the technology of CO2 injection-enhanced coal-bed methane (CO2-ECBM) recovery is still in its infancy with an unclear mechanism. Density functional theory (DFT) calculations were performed to elucidate the mechanism of CO2 adsorption-assisted CH4 desorption (AAD). To simulate coal surfaces, different six-ring aromatic clusters (2 × 2, 3 × 3, 4 × 4, 5 × 5, 6 × 6, and 7 × 7) were used as simplified graphene (Gr) carbon models. The adsorption and desorption of CH4 and/or CO2 on these carbon models were assessed. The results showed that a six-ring aromatic cluster model (4 × 4) can simulate the coal surface with limited approximation. The adsorption of CO2 onto these carbon models was more stable than that in the case of CH4. Further, the adsorption energies of single CH4 and CO2 in the more stable site were -15.58 and -18.16 kJ/mol, respectively. When two molecules (CO2 and CH4) interact with the surface, CO2 compels CH4 to adsorb onto the less stable site, with a resulting significant decrease in the adsorption energy of CH4 onto the surface of the carbon model with pre-adsorbed CO2. The Mulliken charges and electrostatic potentials of CH4 and CO2 adsorbed onto the surface of the carbon model were compared to determine their respective adsorption activities and changes. At the molecular level, our results showed that the adsorption of the injected CO2 promoted the desorption of CH4, the underlying mechanism of CO2-ECBM.

  13. Desorption of H atoms from graphite (0001) using XUV free electron laser pulses

    DEFF Research Database (Denmark)

    Siemer, B.; Olsen, Thomas; Hoger, T.;

    2010-01-01

    , and identifies the highest vibrational state in the adsorbate potential as a major source for the slow atoms. It is evident that multiple electron scattering processes are required for this desorption. A direct electronic excitation of a repulsive hydrogen-carbon bond seems not to be important.......The desorption of neutral H atoms from graphite with femtosecond XUV pulses is reported. The velocity distribution of the atoms peaks at extremely low kinetic energies. A DFT-based electron scattering calculation traces this distribution to desorption out of specific adsorption sites on graphite...

  14. Optical laser-induced CO desorption from Ru(0001) monitored with a free-electron X-ray laser

    DEFF Research Database (Denmark)

    Öberg, H.; Gladh, J.; Dell'Angela, M.

    2015-01-01

    We present density functional theory modeling of time-resolved optical pump/X-ray spectroscopic probe data of CO desorption from Ru(0001). The BEEF van der Waals functional predicts a weakly bound state as a precursor to desorption. The optical pump leads to a near-instantaneous (b100 fs) increas...

  15. High-Resolution Live-Cell Imaging and Analysis by Laser Desorption/Ionization Droplet Delivery Mass Spectrometry.

    Science.gov (United States)

    Lee, Jae Kyoo; Jansson, Erik T; Nam, Hong Gil; Zare, Richard N

    2016-05-17

    We have developed a new ambient-ionization mass spectrometric technique named laser desorption/ionization droplet delivery mass spectrometry (LDIDD-MS). LDIDD-MS permits high-resolution, high-sensitivity imaging of tissue samples as well as measurements of both single-cell apoptosis and live-cell exocytosis. A pulsed (15 Hz) UV laser beam (266 nm) is focused on a surface covered with target analytes to trigger their desorption and ionization. A spray of liquid droplets is simultaneously directed onto the laser-focused surface region to capture the ionized analytes and deliver them to a mass spectrometer. The approach of rapid and effective capturing of molecules after laser desorption/ionization allows the limit of detection for the amino acid lysine to be as low as 2 amol under ambient ionization conditions. Two-dimensional maps of the desorbed/ionized species are recorded by moving the sample on an XY translational stage. The spatial resolution for imaging with LDIDD-MS was determined to be 2.4 μm for an ink-printed pattern and 3 μm for mouse brain tissue. We applied LDIDD-MS to single-cell analysis of apoptotic HEK cells. Differences were observed in the profiles of fatty acids and lipids between healthy HEK cells and those undergoing apoptosis. We observed upregulation of phosphatidylcholine (PC) with a relatively shorter carbon chain length and downregulation of PC with a relatively longer carbon chain length. We also applied LDIDD-MS for a real-time direct measurements of live-cell exocytosis. The catecholamine dopamine and trace amines (phenethylamine and tyramine) were detected from live PC12 cells without damaging them.

  16. Direct Detection of Pharmaceuticals and Personal Care Products from Aqueous Samples with Thermally-Assisted Desorption Electrospray Ionization Mass Spectrometry

    Science.gov (United States)

    Campbell, Ian S.; Ton, Alain T.; Mulligan, Christopher C.

    2011-07-01

    An ambient mass spectrometric method based on desorption electrospray ionization (DESI) has been developed to allow rapid, direct analysis of contaminated water samples, and the technique was evaluated through analysis of a wide array of pharmaceutical and personal care product (PPCP) contaminants. Incorporating direct infusion of aqueous sample and thermal assistance into the source design has allowed low ppt detection limits for the target analytes in drinking water matrices. With this methodology, mass spectral information can be collected in less than 1 min, consuming ~100 μL of total sample. Quantitative ability was also demonstrated without the use of an internal standard, yielding decent linearity and reproducibility. Initial results suggest that this source configuration is resistant to carryover effects and robust towards multi-component samples. The rapid, continuous analysis afforded by this method offers advantages in terms of sample analysis time and throughput over traditional hyphenated mass spectrometric techniques.

  17. Tailor-Made Stable Zr(IV)-Based Metal-Organic Frameworks for Laser Desorption/Ionization Mass Spectrometry Analysis of Small Molecules and Simultaneous Enrichment of Phosphopeptides.

    Science.gov (United States)

    Chen, Lianfang; Ou, Junjie; Wang, Hongwei; Liu, Zhongshan; Ye, Mingliang; Zou, Hanfa

    2016-08-10

    Although thousands of metal-organic frameworks (MOFs) have been fabricated and widely applied in gas storage/separations, adsorption, catalysis, and so on, few kinds of MOFs have been used as adsorption materials while simultaneously serving as matrixes to analyze small molecules for laser desorption/ionization mass spectrometry (LDI-MS). Herein, a new concept is introduced to design and synthesize MOFs as both adsorption materials and matrixes according to the structure of ligands and common matrixes. The proof of concept design was demonstrated by selection of 2,5-pyridinedicarboxylic acid (PDC) and 2,5-dihydroxyterephthalic acid (DHT) as ligands for synthesis of MOFs. Two Zr(IV)-based MOFs of UiO-66-PDC and UiO-66-(OH)2 were synthesized and applied for the first time as new matrixes for analysis of small molecules by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Both of them showed low matrix interferences, high ionization efficiency, and good reproducibility when used as matrixes. A variety of small molecules, including saccharides, amino acids, nucleosides, peptides, alkaline drugs, and natural products, were analyzed. In addition, UiO-66-(OH)2 exhibited potential for application in the quantitative determination of glucose and pyridoxal 5'-phosphate. Furthermore, thanks to its intrinsically large surface area and highly ordered pores, UiO-66-(OH)2 also showed sensitive and specific enrichment of phosphopeptides prior to MS analysis. These results demonstrated that this strategy can be used to efficiently screen tailor-made MOFs as matrixes to analyze small molecules by MALDI-TOF-MS.

  18. Direct analysis of samples by mass spectrometry: From elements to bio-molecules using laser ablation inductively couple plasma mass spectrometry and laser desorption/ionization mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Perdian, David C. [Iowa State Univ., Ames, IA (United States)

    2009-01-01

    Mass spectrometric methods that are able to analyze solid samples or biological materials with little or no sample preparation are invaluable to science as well as society. Fundamental research that has discovered experimental and instrumental parameters that inhibit fractionation effects that occur during the quantification of elemental species in solid samples by laser ablation inductively coupled plasma mass spectrometry is described. Research that determines the effectiveness of novel laser desorption/ionization mass spectrometric methods for the molecular analysis of biological tissues at atmospheric pressure and at high spatial resolution is also described. A spatial resolution is achieved that is able to analyze samples at the single cell level.

  19. A Compact, Solid-State UV (266 nm) Laser System Capable of Burst-Mode Operation for Laser Ablation Desorption Processing

    Science.gov (United States)

    Arevalo, Ricardo, Jr.; Coyle, Barry; Paulios, Demetrios; Stysley, Paul; Feng, Steve; Getty, Stephanie; Binkerhoff, William

    2015-01-01

    Compared to wet chemistry and pyrolysis techniques, in situ laser-based methods of chemical analysis provide an ideal way to characterize precious planetary materials without requiring extensive sample processing. In particular, laser desorption and ablation techniques allow for rapid, reproducible and robust data acquisition over a wide mass range, plus: Quantitative, spatially-resolved measurements of elemental and molecular (organic and inorganic) abundances; Low analytical blanks and limits-of-detection ( ng g-1); and, the destruction of minimal quantities of sample ( g) compared to traditional solution and/or pyrolysis analyses (mg).

  20. Effect of heat treatment on UV-laser-induced positive ion desorption in CaHPO4.2H2O

    Science.gov (United States)

    Kawaguchi, Y.; Dawes, M. L.; Langford, S. C.; Dickinson, J. T.

    We have investigated KrF excimer-laser-induced positive ion desorption from single-crystal brushite (CaHPO4.2H2O), a model biomineral containing water, and we show the effect of heat treatment on ion desorption. Time-of-flight peaks of Ca+ desorption from the heated-cleaved surface are similar to those from as-cleaved ones, but the ion intensity is 2 to 4 orders of magnitude larger. In addition to Ca+, desorption of CaO+, PO+, and P+ from the heated surface is also strongly enhanced. The heated-cleaved surface shows rough, platelet-like fine structures due to recrystallization. Surface defects created during recrystallization strongly couple with the 5 eV photons and dramatically enhance ion desorption.

  1. Functionalization of silicon nanowire arrays by silver nanoparticles for the laser desorption ionization mass spectrometry analysis of vegetable oils.

    Science.gov (United States)

    Picca, Rosaria Anna; Calvano, Cosima Damiana; Lo Faro, Maria Josè; Fazio, Barbara; Trusso, Sebastiano; Ossi, Paolo Maria; Neri, Fortunato; D'Andrea, Cristiano; Irrera, Alessia; Cioffi, Nicola

    2016-09-01

    In this work, novel hybrid nanostructured surfaces, consisting of dense arrays of silicon nanowires (SiNWs) functionalized by Ag nanoparticles (AgNP/SiNWs), were used for the laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS) analysis of some typical unsaturated food components (e.g. squalene, oleic acid) to assess their MS performance. The synthesis of the novel platforms is an easy, cost-effective process based on the maskless wet-etching preparation at room temperature of SiNWs followed by their decoration with AgNPs, produced by pulsed laser deposition. No particular surface pretreatment or addition of organic matrixes/ionizers was necessary. Moreover, oil extracts (e.g. extra virgin olive oil, peanut oil) could be investigated on AgNP/SiNWs surfaces, revealing their different MS profiles. It was shown that such substrates operate at reduced laser energy, typically generating intense silver cluster ions and analyte adducts. A comparison with bare SiNWs was also performed, indicating the importance of AgNP density on NW surface. In this case, desorption/ionization on silicon was invoked as probable LDI mechanism. Finally, the influence of SiNW length and surface composition on MS results was assessed. The combination of typical properties of SiNWs (hydrophobicity, antireflectivity) with ionization ability of metal NPs can be a valid methodology for the further development of nanostructured surfaces in LDI-TOF MS applications. Copyright © 2016 John Wiley & Sons, Ltd.

  2. On the determination of molecular weight distributions of asphaltenes and their aggregates in laser desorption ionization experiments

    Energy Technology Data Exchange (ETDEWEB)

    Hortal, A.R.; Martinez-Haya, B.; Lobato, M.D.; Pedrosa, J.M.; Lago, S. [University of Pablo Olavide, Seville (Spain)

    2006-07-15

    Molecular weight distributions (MWD) of asphaltenes and their aggregates have been investigated in laser desorption ionization (LDI) mass spectrometric experiments. A systematic investigation of the dependence of the measured MIND on the asphaltene sample density and on the laser pulse energy allows the assignment of most probable molecular weights within 300-500 amu and average molecular weights of 800-1000 amu for the monomeric asphaltenes, as well as for the estimation of the contribution from asphaltene clusters in typical LDI measurements. The results serve to reconcile the existing controversy between earlier mass spectrometric characterizations of asphaltenes based on laser desorption techniques by different groups. Furthermore, the MWD measurements performed on particularly dense samples yield an additional differentiated broad band peaking around 9000-10,000 amu and extending over 20,000 amu, not observed previously in LDI experiments, thereby revealing a strong propensity of the asphaltenes to form clusters with specific aggregation numbers, which is in qualitative agreement with previous theoretical predictions and with the interpretation of measurements performed with other techniques.

  3. Millisecond laser machining of transparent materials assisted by nanosecond laser.

    Science.gov (United States)

    Pan, Yunxiang; Zhang, Hongchao; Chen, Jun; Han, Bing; Shen, Zhonghua; Lu, Jian; Ni, Xiaowu

    2015-01-26

    A new form of double pulse composed of a nanosecond laser and a millisecond laser is proposed for laser machining transparent materials. To evaluate its advantages and disadvantages, experimental investigations are carried out and the corresponding results are compared with those of single millisecond laser. The mechanism is discussed from two aspects: material defects and effects of modifications induced by nanosecond laser on thermal stress field during millisecond laser irradiation. It is shown that the modifications of the sample generated by nanosecond laser improves the processing efficiency of subsequent millisecond laser, while limits the eventual size of modified region.

  4. Complications from laser-assisted liposuction performed by noncore practitioners.

    Science.gov (United States)

    Blum, Craig A; Sasser, Charles G S; Kaplan, Jonathan L

    2013-10-01

    Liposuction is one of the most commonly performed aesthetic surgery procedures in the United States, and most plastic surgeons perform suction-assisted, ultrasound-assisted, or power-assisted liposuction. The past decade has seen a growing interest in laser-assisted liposuction (LAL) and the proposed advantages of traditional liposuction methods. However, it is performed by a minority of plastic surgeons. In fact, many LAL providers are not trained in aesthetic practice, and many offer LAL as their only body-contouring procedure. When only one method of body contouring is available to a provider, it may lead to inappropriate patient selection with associated poor outcomes. This report discusses the use of laser liposuction in body contouring and the demographics of those performing liposuction, including LAL. Complications from laser-assisted liposuction performed by noncore practitioners are illustrated.

  5. Eigenstates of Laser-Assisted Scattering in a Noninertial Frame

    Institute of Scientific and Technical Information of China (English)

    FENG Zi-Min; LI Shu-Min

    2005-01-01

    In a rotating noninertial frame, we investigate the eigenstates of the time-dependent problem for electronnucleus scattering assisted by a circularly polarized laser field. Numerical results of probability distribution, quantum potential, and current density are discussed. An approximate expression of scattering cross section for low laser frequency is given.

  6. 21 CFR 884.6200 - Assisted reproduction laser system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Assisted reproduction laser system. 884.6200 Section 884.6200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OBSTETRICAL AND GYNECOLOGICAL DEVICES Assisted Reproduction Devices §...

  7. Process characteristics of fibre-laser-assisted plasma arc welding

    OpenAIRE

    Mahrle, A; SCHNICK, M; Rose, S; Demuth, C; Beyer, E.; Füssel, U

    2011-01-01

    Abstract Experimental and theoretical investigations on fibre-laser assisted plasma arc welding (LAPW) have been performed. Welding experiments were carried out on aluminium and steel sheets. In case of a highly focused laser beam and a separate arrangement of plasma torch and laser beam, high-speed video recordings of the plasma arc and corresponding measurements of the time-dependent arc voltage revealed differences in the process behaviour for both materials. In case of aluminium weldin...

  8. Detection of trace organics in Mars analog samples containing perchlorate by laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Li, Xiang; Danell, Ryan M; Brinckerhoff, William B; Pinnick, Veronica T; van Amerom, Friso; Arevalo, Ricardo D; Getty, Stephanie A; Mahaffy, Paul R; Steininger, Harald; Goesmann, Fred

    2015-02-01

    Evidence from recent Mars missions indicates the presence of perchlorate salts up to 1 wt % level in the near-surface materials. Mixed perchlorates and other oxychlorine species may complicate the detection of organic molecules in bulk martian samples when using pyrolysis techniques. To address this analytical challenge, we report here results of laboratory measurements with laser desorption mass spectrometry, including analyses performed on both commercial and Mars Organic Molecule Analyzer (MOMA) breadboard instruments. We demonstrate that the detection of nonvolatile organics in selected spiked mineral-matrix materials by laser desorption/ionization (LDI) mass spectrometry is not inhibited by the presence of up to 1 wt % perchlorate salt. The organics in the sample are not significantly degraded or combusted in the LDI process, and the parent molecular ion is retained in the mass spectrum. The LDI technique provides distinct potential benefits for the detection of organics in situ on the martian surface and has the potential to aid in the search for signs of life on Mars.

  9. Matrix-free UV-laser desorption/ionization (LDI) mass spectrometric imaging at the single-cell level: distribution of secondary metabolites of Arabidopsis thaliana and Hypericum species.

    Science.gov (United States)

    Hölscher, Dirk; Shroff, Rohit; Knop, Katrin; Gottschaldt, Michael; Crecelius, Anna; Schneider, Bernd; Heckel, David G; Schubert, Ulrich S; Svatos, Ales

    2009-12-01

    The present paper describes matrix-free laser desorption/ionisation mass spectrometric imaging (LDI-MSI) of highly localized UV-absorbing secondary metabolites in plant tissues at single-cell resolution. The scope and limitations of the method are discussed with regard to plants of the genus Hypericum. Naphthodianthrones such as hypericin and pseudohypericin are traceable in dark glands on Hypericum leaves, placenta, stamens and styli; biflavonoids are also traceable in the pollen of this important phytomedical plant. The highest spatial resolution achieved, 10 microm, was much higher than that achieved by commonly used matrix-assisted laser desorption/ionization (MALDI) imaging protocols. The data from imaging experiments were supported by independent LDI-TOF/MS analysis of cryo-sectioned, laser-microdissected and freshly cut plant material. The results confirmed the suitability of combining laser microdissection (LMD) and LDI-TOF/MS or LDI-MSI to analyse localized plant secondary metabolites. Furthermore, Arabidopsis thaliana was analysed to demonstrate the feasibility of LDI-MSI for other commonly occurring compounds such as flavonoids. The organ-specific distribution of kaempferol, quercetin and isorhamnetin, and their glycosides, was imaged at the cellular level.

  10. Assisting Gas Optimization in CO2 Laser Welding

    DEFF Research Database (Denmark)

    Gong, Hui; Olsen, Flemming Ove

    1996-01-01

    are applied with three different flow rates for each of the gases. A number of systematic laboratory experiments is carried out by employing various experimental designs, 33 and 32 Factorial Design. In the experiments a CO2 laser is used to weld thin sheets of mild steel. The welding specimens are evaluated....... The relations between the gas parameters and signal frequencies are studied. Finally the relationships of the gas parameters and weld characteristics are studied. The possibility of using photo diode signals in controlling the gas parameters is discussed.KeywordsCO2 laser welding, assisting gas parameters......High quality laser welding is achieved under the condition of optimizing all process parameters. Assisting gas plays an important role for sound welds. In the conventional welding process assisting gas is used as a shielding gas to prevent that the weld seam oxidates. In the laser welding process...

  11. 应用基质辅助激光解吸电离飞行时间质谱技术建立急性早幼粒细胞白血病微小残留病诊断模型%Establishment of diagnostic model to monitor minimal residual disease of acute promyelocytic leukemia by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    张琳琳; 徐智芳; 覃艳红; 陈秀花; 许艾宁; 任方刚; 王宏伟

    2013-01-01

    目的 比较初诊、完全缓解期急性早幼粒细胞白血病(APL)患者和健康对照者血清中表达有显著差异的蛋白峰,寻找APL及其微小残留病(MRD)的标志蛋白,建立APL MRD诊断模型.方法收集36例初诊APL患者、29例CR期APL患者和32名健康人对照外周血标本,经铜螯合磁珠纯化蛋白后,利用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术进行蛋白质谱检测,采用Flex Analysis 3.0及ClinProTooLs 2.2软件进行分析,构建分类模型并筛选差异蛋白.结果 在质荷比(m/z)1000~10 000范围内,初诊APL患者与健康对照比较有34种血清蛋白质谱峰强度差异有统计学意义(P<0.001),初诊与CR期比较有7种血清蛋白质谱峰强度差异有统计学意义(P<0.001),结果提示m/z为4642、7764和9289质谱峰可以作为APL MRD的标志蛋白峰.利用m/z为4642和9289标志蛋白峰建立APL MRD诊断模型并进行外部盲法验证,8例患者中有6例可以正确诊断.结论 MALDI-TOF MS技术可用于APL患者MRD的动态监测.m/z 4642和9289多肽可能为较好的蛋白标志物.%Objective To screen the potential protein biomarkers in minimal residual disease (MRD) of the acute promyelocytic leukemia (APL) by comparison of differentially expressed serum protein between APL patients at diagnosis and after complete remission (CR) and healthy controls,and to establish and verify a diagnostic model. Methods Serum proteins from 36 cases of primary APL, 29 cases of APL during complete remission and 32 healthy controls were purified by magnetic beads and then analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS). The spectra were analyzed statistically using FlexAnalysisTM and ClinProtTM software. Results Two prediction model of primary APL/healthy control, primary APL/APL CR were developed. Thirty four statistically significant peptide peaks were obtained with the m/z value ranging from 1000 to 10 000 (P<0

  12. Respiratory complications after diode-laser-assisted tonsillotomy.

    Science.gov (United States)

    Fischer, Miloš; Horn, Iris-Susanne; Quante, Mirja; Merkenschlager, Andreas; Schnoor, Jörg; Kaisers, Udo X; Dietz, Andreas; Kluba, Karsten

    2014-08-01

    Children with certain risk factors, such as comorbidities or severe obstructive sleep apnea syndrome (OSAS) are known to require extended postoperative monitoring after adenotonsillectomy. However, there are no recommendations available for diode-laser-assisted tonsillotomy. A retrospective chart review of 96 children who underwent diode-laser-assisted tonsillotomy (07/2011-06/2013) was performed. Data for general and sleep apnea history, power of the applied diode-laser (λ = 940 nm), anesthesia parameters, the presence of postoperative respiratory complications and postoperative healing were evaluated. After initially uncomplicated diode-laser-assisted tonsillotomy, an adjustment of post-anesthesia care was necessary in 16 of 96 patients due to respiratory failure. Respiratory complications were more frequent in younger children (3.1 vs. 4.0 years, p = 0.049, 95 % CI -1.7952 to -0.0048) and in children who suffered from nocturnal apneas (OR = 5.00, p diode-laser power higher than 13 W could be identified as a risk factor for the occurrence of a postoperative oropharyngeal edema (OR = 3.45, p diode-laser-assisted tonsillotomy. We recommend a reduced diode-laser power (<13 W) to reduce oropharyngeal edema.

  13. Laser Assisted Free-Free Transition in Electron - Atom Collision

    Science.gov (United States)

    Sinha, C.; Bhatia, A. K.

    2011-01-01

    Free-free transition is studied for electron-Hydrogen atom system in ground state at very low incident energies in presence of an external homogeneous, monochromatic and linearly polarized laser field. The incident electron is considered to be dressed by the laser in a non perturbative manner by choosing the Volkov solutions in both the channels. The space part of the scattering wave function for the electron is solved numerically by taking into account the effect of electron exchange, short range as well as of the long range interactions. Laser assisted differential as well as elastic total cross sections are calculated for single photon absorption/emission in the soft photon limit, the laser intensity being much less than the atomic field intensity. A strong suppression is noted in the laser assisted cross sections as compared to the field free situations. Significant difference is noted in the singlet and the triplet cross sections.

  14. Layer-by-layer thin film of reduced graphene oxide and gold nanoparticles as an effective sample plate in laser-induced desorption/ionization mass spectrometry.

    Science.gov (United States)

    Kuo, Tsung-Rong; Wang, Di-Yan; Chiu, Yu-Chen; Yeh, Yun-Chieh; Chen, Wei-Ting; Chen, Ching-Hui; Chen, Chun-Wei; Chang, Huan-Cheng; Hu, Cho-Chun; Chen, Chia-Chun

    2014-01-27

    This work demonstrated a simple platform for rapid and effective surface-assisted laser desorption/ionization time-of-flight mass spectrometry (SALDI-TOF MS) measurements based on the layer structure of reduced graphene oxide (rGO) and gold nanoparticles. A multi-layer thin film was fabricated by alternate layer-by-layer depositions of rGO and gold nanoparticles (LBL rGO/AuNP). The flat and clean two-dimensional film was served as the sample plate and also functioned as the matrix in SALDI-TOF MS. By simply one-step deposition of analytes onto the LBL rGO/AuNP sample plate, the MS measurements of various homogeneous samples were ready to execute. The optimization of MS signal was reached by the variation of the layer numbers of rGO and gold nanoparticles. Also, the small molecules including amino acids, carbohydrates and peptides were successfully analyzed in SALDI-TOF MS using the LBL rGO/AuNP sample plate. The results showed that the signal intensity, S N(-1) ratio and reproducibility of SALDI-TOF spectra have been significantly improved in comparison to the uses of gold nanoparticles or α-cyano-4-hydroxy-cinnamic acid (CHCA) as the assisted matrixes. Taking the advantages of the unique properties of rGO and gold nanoparticles, the ready-to-use MS sample plate, which could absorb and dissipate laser energy to analytes quite efficiently and homogeneously, has shown great commercial potentials for MS applications.

  15. Laser-induced desorption of overlayer films off a heated metal substrate

    Science.gov (United States)

    Gu, Xiang; Urbassek, Herbert M.

    2007-02-01

    The temperature-induced desorption of adsorbed overlayer films with thicknesses between 4 and 200 ML off a suddenly heated metal substrate is studied using molecular-dynamics simulation. We observe that the rapid heating vaporizes the surface-near part of the overlayer film. The initial heating-induced thermoelastic pressure and the vapor pressure in the vapor film drive the remaining film as a large relatively cold cluster away from the surface. In our simulations, the material present in the developing vapor film amounts to roughly 2 ML and is quite independent of the overlayer film thickness. For cluster thicknesses beyond 40 ML, the desorption time increases only little with film thickness, while the resulting cluster velocity decreases only slightly.

  16. Femtosecond laser-assisted cataract surgery: A current review

    Directory of Open Access Journals (Sweden)

    Majid Moshirfar

    2011-01-01

    Full Text Available To evaluate the safety, efficacy, advantages, and limitations of femtosecond laser-assisted cataract surgery through a review of the literature. A PubMed search was conducted using topic-appropriate keywords to screen and select articles. Initial research has shown appropriate safety and efficacy of femtosecond laser-assisted cataract surgery, with improvements in anterior capsulotomy, phacofragmentation, and corneal incision. Limitations of these studies include small sample size and short-term follow-up. Cost-benefit analysis has not yet been addressed. Preliminary data for femtosecond laser-assisted cataract surgery shows appropriate safety and efficacy, and possible advantage over conventional cataract surgery. Questions to eventually be answered include comparisons of long-term postoperative complication rates-including infection and visual outcomes-and analysis of contraindications and financial feasibility.

  17. Fractional laser-assisted drug delivery

    DEFF Research Database (Denmark)

    Taudorf, E H; Lerche, C M; Erlendsson, A M

    2016-01-01

    BACKGROUND AND OBJECTIVE: Ablative fractional laser (AFXL) facilitates delivery of topical methotrexate (MTX). This study investigates impact of laser-channel depth on topical MTX-delivery. MATERIALS AND METHODS: MTX (1% [w/v]) diffused for 21 hours through AFXL-exposed porcine skin in in vitro......-thickness skin, biodistribution profiles at specific skin levels, and transdermal permeation. Fluorescence microscopy was used to visualize UVC-activated MTX-fluorescence (254 nm) and semi-quantify MTX distribution in skin. RESULTS: AFXL increased topical MTX-delivery (P ... of coagulation zones (6-47 μm, P ≥ 0.438). CONCLUSION: AFXL greatly increases topical MTX-delivery. Deeper MAZs deliver higher MTX-concentrations than superficial MAZs, which indicates that laser channel depth may be important for topical delivery of hydrophilic molecules. Lasers Surg. Med. 48:519-529, 2016...

  18. Fractional laser-assisted drug delivery

    DEFF Research Database (Denmark)

    Erlendsson, Andrés M; Doukas, Apostolos G; Farinelli, William A

    2016-01-01

    , potentially due to insufficient drug uptake in deeper skin layers. This study sought to investigate a standardized method to actively fill laser-generated channels by altering pressure, vacuum, and pressure (PVP), enquiring its effect on (i) relative filling of individual laser channels; (ii) cutaneous...... deposition and delivery kinetics; (iii) biodistribution and diffusion pattern, estimated by mathematical simulation. METHODS: Franz diffusion chambers (FCs) were used to evaluate the PVP-technique, comparing passive (AFXL) and active (AFXL + PVP) channel filling. A fractional CO2-laser generated superficial...... (225 µm;17.5 mJ/channel) and deep (1200 µm; 130.5 mJ/channel) channels, and PVP was delivered as a 3-minutes cycle of 1 minute pressure (+1.0 atm), 1 minute vacuum (-1.0 atm), and 1 minute pressure (+1.0 atm). Filling of laser channels was visualized with a colored biomarker liquid (n = 12 FCs, n = 588...

  19. Matrices in Analysis of Saccharides by Matrix- assisted Laser Desorption/ionization Time - of- flight Mass Spectrometry%基体辅助激光解吸电离飞行时间质谱分析糖类的基体

    Institute of Scientific and Technical Information of China (English)

    张东娣; 邓慧敏

    2005-01-01

    糖类物质极性高、难挥发、热不稳定,其中多糖还具有相对分子质量分布发散的性质,其质谱表征比较困难.基体辅助激光解吸电离飞行时间质谱(matrix-assistedlazer desorption/ionizationtime-of-flight mass spectrometry,MALDI-TOFMS)灵敏、快速,对杂质的包容性强、分析的质量范围大,能够对糖类尤其是非衍生化糖进行直接分析.近年来,MALDI-TOFMS的发展已使糖类物质的分析达到一个新的水平.由于基体在MALDI-TOFMS分析中起着至关重要的作用,而研制开发和使用各种有效新基体也一直是MALDI-TOFMS分析的聚焦点,本文综述近10年MALDI-TOFMS分析糖类所用的基体.

  20. Characterization of aromaticity in analogues of titan's atmospheric aerosols with two-step laser desorption ionization mass spectrometry

    CERN Document Server

    Mahjoub, Ahmed; Carrasco, Nathalie; Benilan, Yves; Cernogora, Guy; Szopa, Cyril; Gazeau, Marie-Claire

    2016-01-01

    The role of polycyclic aromatic hydrocarbons (PAH) and Nitrogen containing PAH (PANH) as intermediates of aerosol production in the atmosphere of Titan has been a subject of controversy for a long time. An analysis of the atmospheric emission band observed by the Visible and Infrared Mapping Spectrometer (VIMS) at 3.28 micrometer suggests the presence of neutral polycyclic aromatic species in the upper atmosphere of Titan. These molecules are seen as the counter part of negative and positive aromatics ions suspected by the Plasma Spectrometer onboard the Cassini spacecraft, but the low resolution of the instrument hinders any molecular speciation. In this work we investigate the specific aromatic content of Titan's atmospheric aerosols through laboratory simulations. We report here the selective detection of aromatic compounds in tholins, Titan's aerosol analogues, produced with a capacitively coupled plasma in a N2:CH4 95:5 gas mixture. For this purpose, Two-Step Laser Desorption Ionization Time-of-Flight Ma...

  1. Aptamer-conjugated multifunctional nanoflowers as a platform for targeting, capture, and detection in laser desorption ionization mass spectrometry.

    Science.gov (United States)

    Ocsoy, Ismail; Gulbakan, Basri; Shukoor, Mohammed Ibrahim; Xiong, Xiangling; Chen, Tao; Powell, David H; Tan, Weihong

    2013-01-22

    Although many different nanomaterials have been tested as substrates for laser desorption and ionization mass spectrometry (LDI-MS), this emerging field still requires more efficient multifuncional nanomaterials for targeting, enrichment, and detection. Here, we report the use of gold manganese oxide (Au@MnO) hybrid nanoflowers as an efficient matrix for LDI-MS. The nanoflowers were also functionalized with two different aptamers to target cancer cells and capture adenosine triphosphate (ATP). These nanoflowers were successfully used for metabolite extraction from cancer cell lysates. Thus, in one system, our multifunctional nanoflowers can (1) act as an ionization substrate for mass spectrometry, (2) target cancer cells, and (3) detect and analyze metabolites from cancer cells.

  2. Aptamer Conjugated Multifunctional Nanoflowers as a Platform for Targeting, Capture and Detection in Laser Desorption Ionization Mass Spectrometry

    Science.gov (United States)

    Ocsoy, Ismail; Gulbakan, Basri; Shukoor, Mohammed Ibrahim; Xiong, Xiangling; Chen, Tao; Powell, David H.; Tan, Weihong

    2013-01-01

    Although many different nanomaterials have been tested as substrates for laser desorption and ionization mass spectrometry (LDI-MS), this emerging field still requires more efficient multifuncional nanomaterials for targeting, enrichment and detection. Here, we report the use of gold-manganese oxide (Au@MnO) hybrid nanoflowers as an efficient matrix for LDI–MS. The nanoflowers were also functionalized with two different aptamers to target cancer cells and capture adenosine triphosphate (ATP), respectively. These nanoflowers were successfully used for metabolite extraction from cancer cell lysates. Thus, in one system, our multifunctional nanoflowers can 1) act as an ionization substrate for mass spectrometry, 2) target cancer cells, and 3) detect and analyze metabolites from cancer cells. PMID:23211039

  3. Carbamazepine in municipal wastewater and wastewater sludge: ultrafast quantification by laser diode thermal desorption-atmospheric pressure chemical ionization coupled with tandem mass spectrometry.

    Science.gov (United States)

    Mohapatra, D P; Brar, S K; Tyagi, R D; Picard, P; Surampalli, R Y

    2012-09-15

    In this study, the distribution of the anti-epileptic drug carbamazepine (CBZ) in wastewater (WW) and aqueous and solid phases of wastewater sludge (WWS) was carried out. A rapid and reliable method enabling high-throughput sample analysis for quicker data generation, detection, and monitoring of CBZ in WW and WWS was developed and validated. The ultrafast method (15s per sample) is based on the laser diode thermal desorption-atmospheric pressure chemical ionization (LDTD-APCI) coupled to tandem mass spectrometry (MS/MS). The optimization of instrumental parameters and method application for environmental analysis are presented. The performance of the novel method was evaluated by estimation of extraction recovery, linearity, precision and detection limit. The method detection limits was 12 ng L(-1) in WW and 3.4 ng g(-1) in WWS. The intra- and inter-day precisions were 8% and 11% in WW and 6% and 9% in WWS, respectively. Furthermore, three extraction methods, ultrasonic extraction (USE), microwave-assisted extraction (MAE) and accelerated solvent extraction (ASE) with three different solvent condition such as methanol, acetone and acetonitrile:ethyle acetate (5:1, v/v) were compared on the basis of procedural blank and method recovery. Overall, ASE showed the best extraction efficiency with methanol as compared to USE and MAE. Furthermore, the quantification of CBZ in WW and WWS samples showed the presence of contaminant in all stages of the treatment plant.

  4. Laser-Assisted Removal of Aspirated Thumbtacks by Flexible Bronchoscopy

    Directory of Open Access Journals (Sweden)

    Oren Fruchter

    2010-01-01

    Objectives. To describe a technique for removal of sharp aspirated metal objects employing laser through flexible bronchoscopy. Methods. We report two patients in whom a new technique for removal of sharp aspirated metal objects utilizing Nd-Yag laser flexible bronchoscopy was used. Results. Successful and uncomplicated removal of the aspirated thumbpack by flexible bronchoscopy under conscious sedation was accomplished in the two patients described. Both patients were discharged within 24 hours. Conclusions. In patients with aspirated thumbtack laser-assisted breakage of the object through flexible bronchoscopy may obviate the need for rigid bronchoscopy or thoracotomy.

  5. Frequency Comb Assisted Broadband Precision Spectroscopy with Cascaded Diode Lasers

    CERN Document Server

    Liu, Junqiu; Pfeiffer, Martin H P; Kordts, Arne; Kamel, Ayman N; Guo, Hairun; Geiselmann, Michael; Kippenberg, Tobias J

    2016-01-01

    Frequency comb assisted diode laser spectroscopy, employing both the accuracy of an optical frequency comb and the broad wavelength tuning range of a tunable diode laser, has been widely used in many applications. In this letter we present a novel method using cascaded frequency agile diode lasers, which allows extending the measurement bandwidth to 37.4 THz (1355 to 1630 nm) at MHz resolution with scanning speeds above 1 THz/s. It is demonstrated as a useful tool to characterize a broadband spectrum for molecular spectroscopy and in particular it enables to characterize the dispersion of integrated microresonators up to the fourth order.

  6. Laser and optical system for laser assisted hydrogen ion beam stripping at SNS

    Science.gov (United States)

    Liu, Y.; Rakhman, A.; Menshov, A.; Webster, A.; Gorlov, T.; Aleksandrov, A.; Cousineau, S.

    2017-03-01

    Recently, a high-efficiency laser assisted hydrogen ion (H-) beam stripping was successfully carried out in the Spallation Neutron Source (SNS) accelerator. The experiment was not only an important step toward foil-less H- stripping for charge exchange injection, it also set up a first example of using megawatt ultraviolet (UV) laser source in an operational high power proton accelerator facility. This paper reports in detail the design, installation, and commissioning result of a macro-pulsed multi-megawatt UV laser system and laser beam transport line for the laser stripping experiment.

  7. Discrimination between bacterial spore types using time-of-flight mass spectrometry and matrix-free infrared laser desorption and ionization.

    Science.gov (United States)

    Ullom, J N; Frank, M; Gard, E E; Horn, J M; Labov, S E; Langry, K; Magnotta, F; Stanion, K A; Hack, C A; Benner, W H

    2001-05-15

    We demonstrate that molecular ions with mass-to-charge ratios (m/z) ranging from a few hundred to 19 050 can be desorbed from whole bacterial spores using infrared laser desorption and no chemical matrix. We have measured the mass of these ions using time-of-flight mass spectrometry and we observe that different ions are desorbed from spores of Bacillus cereus, Bacillus thuringiensis, Bacillus subtilis, and Bacillus niger. Our results raise the possibility of identifying microorganisms using mass spectrometry without conventional sample preparation techniques such as the addition of a matrix. We have measured the dependence of the ion yield from B. subtilis on desorption wavelength over the range 3.05-3.8 microm, and we observe the best results at 3.05 microm. We have also generated mass spectra from whole spores using 337-nm ultraviolet laser desorption, and we find that these spectra are inferior to spectra generated with infrared desorption. Since aerosol analysis is a natural application for matrix-free desorption, we have measured mass spectra from materials such as ragweed pollen and road dust that are likely to form a background to microbial aerosols. We find that these materials are readily differentiated from bacterial spores.

  8. Lipase biofilm deposited by Matrix Assisted Pulsed Laser Evaporation technique

    Energy Technology Data Exchange (ETDEWEB)

    Aronne, Antonio [Department of Chemical Engineering, Materials and Industrial Production, University of Naples “Federico II”, Napoli (Italy); Bloisi, Francesco, E-mail: bloisi@na.infn.it [SPIN – CNR, Naples (Italy); Department of Physics, University of Naples “Federico II”, Napoli (Italy); Calabria, Raffaela; Califano, Valeria [Istituto Motori – CNR, Naples (Italy); Depero, Laura E. [Department of Mechanical and Industrial Engineering, University of Brescia, Brescia (Italy); Fanelli, Esther [Department of Chemical Engineering, Materials and Industrial Production, University of Naples “Federico II”, Napoli (Italy); Federici, Stefania [Department of Mechanical and Industrial Engineering, University of Brescia, Brescia (Italy); Massoli, Patrizio [Istituto Motori – CNR, Naples (Italy); Vicari, Luciano R.M. [SPIN – CNR, Naples (Italy); Department of Physics, University of Naples “Federico II”, Napoli (Italy)

    2015-05-01

    Highlights: • A lipase film was deposited with Matrix Assisted Pulsed Laser Evaporation technique. • FTIR spectra show that laser irradiation do not damage lipase molecule. • Laser fluence controls the characteristics of complex structure generated by MAPLE. - Abstract: Lipase is an enzyme that finds application in biodiesel production and for detection of esters and triglycerides in biosensors. Matrix Assisted Pulsed Laser Evaporation (MAPLE), a technique derived from Pulsed Laser Deposition (PLD) for deposition of undamaged biomolecules or polymers, is characterized by the use of a frozen target obtained from a solution/suspension of the guest material (to be deposited) in a volatile matrix (solvent). The presence of the solvent avoids or at least reduces the potential damage of guest molecules by laser radiation but only the guest material reaches the substrate in an essentially solvent-free deposition. MAPLE can be used for enzymes immobilization, essential for industrial application, allowing the development of continuous processes, an easier separation of products, the reuse of the catalyst and, in some cases, enhancing enzyme properties (pH, temperature stability, etc.) and catalytic activity in non-aqueous media. Here we show that MAPLE technique can be used to deposit undamaged lipase and that the complex structure (due to droplets generated during extraction from target) of the deposited material can be controlled by changing the laser beam fluence.

  9. Laser-assisted deposition of thin C60 films

    DEFF Research Database (Denmark)

    Schou, Jørgen; Canulescu, Stela; Fæster, Søren

    . However, organic materials are usually not well suited for direct laser irradiation, since the organic molecules may suffer from fragmentation by the laser light. We have, therefore, explored the possible fragmentation of organic molecules by attempting to produce thin films of C60 which is a strongly...... bound carbon molecule with a well-defined mass (M = 720 amu) and therefore a good, organic test molecule. C60 fullerene thin films of average thickness of more than 100 nm was produced in vacuum by matrix-assisted pulsed laser evaporation (MAPLE). A 355 nm Nd:YAG laser was directed onto a frozen target...... of the matrix material, anisole, with a concentration of 0.67 wt% C60. At laser fluences below 1.5 J/cm2, a dominant fraction of the film molecules are C60 transferred to the substrate without any fragmentation. High-resolution SEM images of MAPLE deposited films reveal large circular features on the surface...

  10. Synthesis of Ag@Silica Nanoparticles by Assisted Laser Ablation

    Science.gov (United States)

    González-Castillo, Jr.; Rodriguez, E.; Jimenez-Villar, E.; Rodríguez, D.; Salomon-García, I.; de Sá, Gilberto F.; García-Fernández, T.; Almeida, DB; Cesar, CL; Johnes, R.; Ibarra, Juana C.

    2015-10-01

    This paper reports the synthesis of silver nanoparticles coated with porous silica (Ag@Silica NPs) using an assisted laser ablation method. This method is a chemical synthesis where one of the reagents (the reducer agent) is introduced in nanometer form by laser ablation of a solid target submerged in an aqueous solution. In a first step, a silicon wafer immersed in water solution was laser ablated for several minutes. Subsequently, an AgNO3 aliquot was added to the aqueous solution. The redox reaction between the silver ions and ablation products leads to a colloidal suspension of core-shell Ag@Silica NPs. The influence of the laser pulse energy, laser wavelength, ablation time, and Ag+ concentration on the size and optical properties of the Ag@Silica NPs was investigated. Furthermore, the colloidal suspensions were studied by UV-VIS-NIR spectroscopy, X-Ray diffraction, and high-resolution transmission electron microscopy (HRTEM).

  11. Fractional CO(2) laser-assisted drug delivery

    DEFF Research Database (Denmark)

    Haedersdal, Merete; Sakamoto, Fernanda H; Farinelli, William A

    2010-01-01

    Ablative fractional resurfacing (AFR) creates vertical channels that might assist the delivery of topically applied drugs into skin. The purpose of this study was to evaluate drug delivery by CO(2) laser AFR using methyl 5-aminolevulinate (MAL), a porphyrin precursor, as a test drug....

  12. Femtosecond Laser Desorption of Thin Polymer Films from a Dielectric Surface

    Directory of Open Access Journals (Sweden)

    Mercadier L.

    2013-11-01

    Full Text Available We desorb polymer films from fused silica with a femtosecond laser and characterize the results by atomic force microscopy. Our study as a function of beam geometry and energy reveals two ways of achieving spatially controlled nanodesorption.

  13. Ultrafast Desorption by Impulsive Vibrational Excitation (DIVE). Applications in laser surgery, mass spectrometry and towards ultimate limits in biodiagnosis

    Energy Technology Data Exchange (ETDEWEB)

    Ren, Ling

    2015-07-15

    The prospects for minimally invasive surgery, spatial imaging with mass spectrometry and rapid high throughput biodiagnosis require new means of tissue incision and biomolecule extraction with conserved molecular structure. Towards this aim, a laser ablation process is utilized in this dissertation, which is capable of performing precise tissue incision with minimal collateral damage and extracting intact biological entities with conserved biological functions. The method is based on the recently developed Picosecond Infrared Laser (PIRL) designed to excite selectively the water vibrational modes under the condition of ultrafast Desorption by Impulsive Vibrational Excitation (DIVE). The basic concept is that the selectively excited water molecules act as propellant to ablate whole biological complexes into the plume, faster than any thermal deleterious effect or fragmentation that would mask molecular identities.The PIRL ablation under DIVE condition is applied for the first time to six types of ocular tissues, rendering precise and minimally invasive incisions in a well-controlled and reproducible way. An eminent demonstration is the contact-free and applanation-free corneal trephination with the PIRL. Mass spectrometry and other analytical techniques show that great abundance of proteins with various molecular weights are extracted from the tissue by the PIRL ablation, and that fragmentation or other chemical alternation does not occur to the proteins in the ablation plume. With various microscope imaging and biochemical analysis methods, nano-scale single protein molecules, viruses and cells in the ablation plume are found to be morphologically and functionally identical to their corresponding controls. The PIRL ablation provides a new means to push the frontiers of laser surgery in ophthalmology and can be applied to resolve chemical activities in situ and in vivo. The most important finding is the conserved nature of the extracted biological entities

  14. Laser-assisted fabrication of highly viscous alginate microsphere

    Science.gov (United States)

    Lin, Yafu; Huang, Yong

    2011-04-01

    Encapsulated microspheres have been widely used in various biomedical applications. However, fabrication of encapsulated microspheres from highly viscous materials has always been a manufacturing challenge. The objective of this study is to explore a novel metallic foil-assisted laser-induced forward transfer (LIFT), a laser-assisted fabrication technique, to make encapsulated microspheres using high sodium alginate concentration solutions. The proposed four-layer approach includes a quartz disk, a sacrificial and adhesive layer, a metallic foil, and a transferred suspension layer. It is found that the proposed four-layer modified LIFT approach provides a promising fabrication technology for making of bead-encapsulated microspheres from highly viscous solutions. During the process, the microsphere only can be formed if the direct-writing height is larger than the critical direct-writing height; otherwise, tail structured droplets are formed; and the encapsulated microsphere diameter linearly increases with the laser fluence and decreases with the sodium alginate concentration.

  15. Laser-assisted machining of difficult-to-machine materials

    Energy Technology Data Exchange (ETDEWEB)

    Incropera, F.P.; Rozzi, J.C.; Pfefferkorn, F.E.; Lei, S.; Shin, Y.C.

    1999-07-01

    Laser-assisted machining (LAM) is a hybrid process for which a difficult-to-machine material, such as a ceramic or super alloy, is irradiated by a laser source prior to material removal by a cutting tool. The process has the potential to significantly increase material removal rates, as well as to improve the geometry and properties of the finished work piece. Features and limitations of theoretical and experimental procedures for determining the transient thermal response of a work piece during LAM are described, and representative results are presented for laser-assisted turning of sintered silicon nitride. Significant physical trends are revealed by the calculations, as are guidelines for the selection of appropriate operating conditions.

  16. Atmospheric pressure laser-induced acoustic desorption chemical ionization Fourier transform ion cyclotron resonance mass spectrometry for the analysis of complex mixtures.

    Science.gov (United States)

    Nyadong, Leonard; McKenna, Amy M; Hendrickson, Christopher L; Rodgers, Ryan P; Marshall, Alan G

    2011-03-01

    We present a novel nonresonant laser-based matrix-free atmospheric pressure ionization technique, atmospheric pressure laser-induced acoustic desorption chemical ionization (AP/LIAD-CI). The technique decouples analyte desorption from subsequent ionization by reagent ions generated from a corona discharge initiated in ambient air or in the presence of vaporized toluene as a CI dopant at room temperature. Analyte desorption is initiated by a shock wave induced in a titanium foil coated with electrosprayed sample, irradiated from the rear side by high-energy laser pulses. The technique enables facile and independent optimization of the analyte desorption, ionization, and sampling events, for coupling to any mass analyzer with an AP interface. Moreover, the generated analyte ions are efficiently thermalized by collisions with atmospheric gases, thereby reducing fragmentation. We have coupled AP/LIAD-CI to ultrahigh-resolution FT-ICR MS to generate predominantly [M + H](+) or M(+•) ions to resolve and identify thousands of elemental compositions from organic mixtures as complex as petroleum crude oil distillates. Finally, we have optimized the AP/LIAD CI process and investigated ionization mechanisms by systematic variation of placement of the sample, placement of the corona discharge needle, discharge current, gas flow rate, and inclusion of toluene as a dopant.

  17. Coupling Laser Diode Thermal Desorption with Acoustic Sample Deposition to Improve Throughput of Mass Spectrometry-Based Screening.

    Science.gov (United States)

    Haarhoff, Zuzana; Wagner, Andrew; Picard, Pierre; Drexler, Dieter M; Zvyaga, Tatyana; Shou, Wilson

    2016-02-01

    The move toward label-free screening in drug discovery has increased the demand for mass spectrometry (MS)-based analysis. Here we investigated the approach of coupling acoustic sample deposition (ASD) with laser diode thermal desorption (LDTD)-tandem mass spectrometry (MS/MS). We assessed its use in a cytochrome P450 (CYP) inhibition assay, where a decrease in metabolite formation signifies CYP inhibition. Metabolite levels for 3 CYP isoforms were measured as CYP3A4-1'-OH-midazolam, CYP2D6-dextrorphan, and CYP2C9-4'-OH-diclofenac. After incubation, samples (100 nL) were acoustically deposited onto a stainless steel 384-LazWell plate, then desorbed by an infrared laser directly from the plate surface into the gas phase, ionized by atmospheric pressure chemical ionization (APCI), and analyzed by MS/MS. Using this method, we achieved a sample analysis speed of 2.14 s/well, with bioanalytical performance comparable to the current online solid-phase extraction (SPE)-based MS method. An even faster readout speed was achieved when postreaction sample multiplexing was applied, where three reaction samples, one for each CYP, were transferred into the same well of the LazWell plate. In summary, LDTD coupled with acoustic sample deposition and multiplexing significantly decreased analysis time to 0.7 s/sample, making this MS-based approach feasible to support high-throughput screening (HTS) assays.

  18. UV laser induced desorption of CsI and RbI ion clusters

    Science.gov (United States)

    Fernández-Lima, F. A.; Ponciano, C. R.; Filho, H. D. Fonseca; Pedrero, E.; Chaer Nascimento, M. A.; da Silveira, E. F.

    2006-09-01

    Experimental results of laser sputtering of cesium and rubidium iodide secondary ions are presented. A TOF mass spectrometer, operating in linear mode, continuous extraction for positive or negative ions, was used for the analysis of (CsI) nCs +, (CsI) nI -, (RbI) nRb + and (RbI) nI - ion emission as a function of the laser irradiance. Experimental data show that the cluster ion emission yields decrease exponentially with n, for all the laser irradiances applied. Theoretical analysis of the clusters structure was performed using density functional theory at the B3LYP/LACV3P level, for the positive and negative cluster series. A quasi-equilibrium evolution of the clusters is proposed to extract a parameter characteristic of the cluster recombination process: the effective temperature. The hypothesis of the atomic species' recombination (during the expansion of a high density highly ionized cloud) leading to cluster formation is confirmed to some extent in a second set of experiments: the UV laser ablation of a mixed and non-mixed cesium iodide and potassium bromide targets. These experiments show that the emission yields contain contributions from both the recombination process and from the sample stoichiometry, even for high laser irradiances.

  19. Molecular dynamics investigation of desorption and ion separation following picosecond infrared laser (PIRL) ablation of an ionic aqueous protein solution.

    Science.gov (United States)

    Zou, J; Wu, C; Robertson, W D; Zhigilei, L V; Miller, R J D

    2016-11-28

    Molecular dynamics simulations were performed to characterize the ablation process induced by a picosecond infrared laser (PIRL) operating in the regime of desorption by impulsive vibrational excitation (DIVE) of a model peptide (lysozyme)/counter-ion system in aqueous solution. The simulations were performed for ablation under typical experimental conditions found within a time-of-flight mass spectrometer (TOF-MS), that is in vacuum with an applied electric field (E = ± 10(7) V/m), for up to 2 ns post-ablation and compared to the standard PIRL-DIVE ablation condition (E = 0 V/m). Further, a simulation of ablation under an extreme field condition (E = 10(10) V/m) was performed for comparison to extend the effective dynamic range of the effect of the field on charge separation. The results show that the plume dynamics were retained under a typical TOF-MS condition within the first 1 ns of ablation. Efficient desorption was observed with more than 90% of water molecules interacting with lysozyme stripped off within 1 ns post-ablation. The processes of ablation and desolvation of analytes were shown to be independent of the applied electric field and thus decoupled from the ion separation process. Unlike under the extreme field conditions, the electric field inside a typical TOF-MS was shown to modify the ions' motion over a longer time and in a soft manner with no enhancement to fragmentation observed as compared to the standard PIRL-DIVE. The study indicates that the PIRL-DIVE ablation mechanism could be used as a new, intrinsically versatile, and highly sensitive ion source for quantitative mass spectrometry.

  20. Molecular dynamics investigation of desorption and ion separation following picosecond infrared laser (PIRL) ablation of an ionic aqueous protein solution

    Science.gov (United States)

    Zou, J.; Wu, C.; Robertson, W. D.; Zhigilei, L. V.; Miller, R. J. D.

    2016-11-01

    Molecular dynamics simulations were performed to characterize the ablation process induced by a picosecond infrared laser (PIRL) operating in the regime of desorption by impulsive vibrational excitation (DIVE) of a model peptide (lysozyme)/counter-ion system in aqueous solution. The simulations were performed for ablation under typical experimental conditions found within a time-of-flight mass spectrometer (TOF-MS), that is in vacuum with an applied electric field (E = ± 107 V/m), for up to 2 ns post-ablation and compared to the standard PIRL-DIVE ablation condition (E = 0 V/m). Further, a simulation of ablation under an extreme field condition (E = 1010 V/m) was performed for comparison to extend the effective dynamic range of the effect of the field on charge separation. The results show that the plume dynamics were retained under a typical TOF-MS condition within the first 1 ns of ablation. Efficient desorption was observed with more than 90% of water molecules interacting with lysozyme stripped off within 1 ns post-ablation. The processes of ablation and desolvation of analytes were shown to be independent of the applied electric field and thus decoupled from the ion separation process. Unlike under the extreme field conditions, the electric field inside a typical TOF-MS was shown to modify the ions' motion over a longer time and in a soft manner with no enhancement to fragmentation observed as compared to the standard PIRL-DIVE. The study indicates that the PIRL-DIVE ablation mechanism could be used as a new, intrinsically versatile, and highly sensitive ion source for quantitative mass spectrometry.

  1. Diode Laser Assisted Filament Winding of Thermoplastic Matrix Composites

    Directory of Open Access Journals (Sweden)

    Claudia Prosperi

    2010-01-01

    Full Text Available A new consolidation method for the laser-assisted filament winding of thermoplastic prepregs is discussed: for the first time a diode laser is used, as well as long glass fiber reinforced polypropylene prepregs. A consolidation apparatus was built by means of a CNC motion table, a stepper motor and a simple tensioner. Preliminary tests were performed in a hoop winding configuration: only the winding speed was changed, and all the other process parameters (laser power, distance from the laser focus, consolidation force were kept constant. Small wound rings with an internal diameter of 25 mm were produced and compression tests were carried out to evaluate the composite agglomeration in dependence of the winding speed. At lower winding speeds, a stronginterpenetration of adjacent layers was observed.

  2. Tailored nanopost arrays (NAPA) for laser desorption ionization in mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Vertes, Akos; Walker, Bennett N.; Stolee, Jessica A.; Retterer, Scott T.

    2016-11-08

    The production and use of semiconducting nanopost arrays made by nanofabrication is described herein. These nanopost arrays (NAPA) provide improved laser ionization yields and controllable fragmentation with switching or modulation capabilities for mass spectrometric detection and identification of samples deposited on them.

  3. Pulsed laser-assisted machining of Inconel 718 superalloy

    Science.gov (United States)

    Azhdari Tadavani, Soheila; Shoja Razavi, Reza; Vafaei, Reza

    2017-01-01

    Nickel-based superalloys including Inconel 718(IN718) are widely used in aerospace industries due to their superior high temperature strength, toughness, and corrosion resistance. These alloys are difficult to machine mainly because of their low thermal conductivity and high work hardening rate, which cause steep temperature gradient and high cutting forces at the tool edge. The application of laser assisted machining is the subject of many new researches since shear forces; surface coarsening and tool wear are reduced. The aim of this investigation was to evaluate laser assisted machining behavior of a 718 Inconel superalloy from the view point of machining specific energy, surface roughness, tool wear and chip appearance. Experimental apparatuses used included optical and scanning electron microscopy, spark emission spectroscopy, and EDS analysis. The results indicated that increasing the temperature to about 540 °C just ahead of primary shear zone, can result in 35% reduction of machining specific energy, in comparison with conventional machining. Furthermore, surface coarsening and tool wear were reduced by 22% and 23% respectively. Flank wear was the main deteriorating factor on cutting tools during laser assisted machining. SEM micrographs indicated that increase in temperature has no noticeable effect on finished workpiece surface. Analysis of variance obtained from regression analysis indicated that frequency of laser beam has the most influential effect on temperature. The optimum conditions for laser assisted machining of 718 superalloy is suggested as follows: 80 Hz frequency, 400 W power, 24 m/min cutting speed, and 0.052 mm/rev feed rate along with 540 °C temperature, 2.51 J/mm2 machining specific energy and 130 N cutting force.

  4. Laser-cooling-assisted mass spectrometry

    CERN Document Server

    Schneider, Christian; Chen, Kuang; Sullivan, Scott T; Hudson, Eric R

    2014-01-01

    Mass spectrometry is used in a wide range of scientific disciplines including proteomics, pharmaceutics, forensics, and fundamental physics and chemistry. Given this ubiquity, there is a worldwide effort to improve the efficiency and resolution of mass spectrometers. However, the performance of all techniques is ultimately limited by the initial phase-space distribution of the molecules being analyzed. Here, we dramatically reduce the width of this initial phase-space distribution by sympathetically cooling the input molecules with laser-cooled, co-trapped atomic ions, improving both the mass resolution and detection efficiency of a time-of-flight mass spectrometer by over an order of magnitude. Detailed molecular dynamics simulations verify the technique and aid with evaluating its effectiveness. Our technique appears to be applicable to other types of mass spectrometers.

  5. Process characteristics of fibre-laser-assisted plasma arc welding

    Energy Technology Data Exchange (ETDEWEB)

    Mahrle, A; Schnick, M; Rose, S; Demuth, C; Beyer, E; Fuessel, U, E-mail: achim.mahrle@iws.fraunhofer.de [Dresden University of Technology, Institute of Surface and Manufacturing Technology, PO Box, D-01062 Dresden (Germany)

    2011-08-31

    Experimental and theoretical investigations on fibre-laser-assisted plasma arc welding (LAPW) were performed. Welding experiments were carried out on aluminium and steel sheets. In the case of a highly focused laser beam and a separate arrangement of plasma torch and laser beam, high-speed video recordings of the plasma arc and corresponding measurements of the time-dependent arc voltage revealed differences in the process behaviour for both materials. In the case of aluminium welding, a sharp decline in arc voltage and stabilization and guiding of the anodic arc root was observed whereas in steel welding the arc voltage was slightly increased after the laser beam was switched on. However, significant improvement of the melting efficiency with the combined action of plasma arc and laser beam was achieved for both types of material. Theoretical results of additional numerical simulations of the arc behaviour suggest that the properties of the arc plasma are mainly influenced not by a direct interaction with the laser radiation but by the laser-induced evaporation of metal. Arc stabilization with increased current densities is predicted for moderate rates of evaporated metal only whereas metal vapour rates above a certain threshold causes a destabilization of the arc and reduced current densities along the arc axis.

  6. Two-Step Resonance-Enhanced Desorption Laser Mass Spectrometry for In Situ Analysis of Organic-Rich Environments

    Science.gov (United States)

    Getty, S. A.; Grubisic, A.; Uckert, K.; Li, X.; Cornish, T.; Cook, J. E.; Brinckerhoff, W. B.

    2016-01-01

    A wide diversity of planetary surfaces in the solar system represent high priority targets for in situ compositional and contextual analysis as part of future missions. The planned mission portfolio will inform our knowledge of the chemistry at play on Mars, icy moons, comets, and primitive asteroids, which can lead to advances in our understanding of the interplay between inorganic and organic building blocks that led to the evolution of habitable environments on Earth and beyond. In many of these environments, the presence of water or aqueously altered mineralogy is an important indicator of habitable environments that are present or may have been present in the past. As a result, the search for complex organic chemistry that may imply the presence of a feedstock, if not an inventory of biosignatures, is naturally aligned with targeted analyses of water-rich surface materials. Here we describe the two-step laser mass spectrometry (L2MS) analytical technique that has seen broad application in the study of organics in meteoritic samples, now demonstrated to be compatible with an in situ investigation with technique improvements to target high priority planetary environments as part of a future scientific payload. An ultraviolet (UV) pulsed laser is used in previous and current embodiments of laser desorption/ionization mass spectrometry (LDMS) to produce ionized species traceable to the mineral and organic composition of a planetary surface sample. L2MS, an advanced technique in laser mass spectrometry, is selective to the aromatic organic fraction of a complex sample, which can provide additional sensitivity and confidence in the detection of specific compound structures. Use of a compact two-step laser mass spectrometer prototype has been previously reported to provide specificity to key aromatic species, such as PAHs, nucleobases, and certain amino acids. Recent improvements in this technique have focused on the interaction between the mineral matrix and the

  7. Laser assisted Breit-Wheeler and Schwinger processes

    OpenAIRE

    Nousch, T.; Otto, A.(European Organization for Nuclear Research (CERN), Geneva, Switzerland); Seipt, D.; Kämpfer, B.; Titov, A. I.; Blaschke, D.; Panferov, A. D.; Smolyansky, S. A.

    2016-01-01

    The assistance of an intense optical laser pulse on electron-positron pair production by the Breit-Wheeler and Schwinger processes in XFEL fields is analyzed. The impact of a laser beam on high-energy photon collisions with XFEL photons consists in a phase space redistribution of the pairs emerging in the Breit-Wheeler sub-process. We provide numerical examples of the differential cross section for parameters related to the European XFEL. Analogously, the Schwinger type pair production in pul...

  8. Determination of metformin in mouse, rat, dog and human plasma samples by laser diode thermal desorption/atmospheric pressure chemical ionization tandem mass spectrometry.

    Science.gov (United States)

    Swales, John G; Gallagher, Richard; Peter, Raimund M

    2010-11-02

    A simple, rapid and robust high-throughput assay for the quantitative analysis of metformin in plasma from different species using laser diode thermal desorption interfaced with atmospheric chemical pressure ionization tandem mass spectrometry (LDTD-APCI-MSMS) was developed for use in a pharmaceutical discovery environment. In order to minimize sample preparation a generic protein precipitation method was used to extract metformin from the plasma. Laser diode thermal desorption is a relatively new sample introduction method, the optimization of the instrumental parameters are presented. The method was successfully applied to spiked mouse, rat, dog and human plasma samples and was subsequently used to determine the oral pharmacokinetics of metformin after dosing to male rats in order to support drug discovery projects. The deviations for intra-assay accuracy and precision across the four species were less than 30% at all calibration and quality control levels.

  9. The effects of laser assisted hatching on pregnancy rates

    OpenAIRE

    Alireza Ghannadi; Marjaneh Kazerooni; Fatemeh Jamalzadeh; Sahar Amiri; Parifar Rostami; Forouzan Absalan

    2011-01-01

    Background: For infertile women aged over 35 years, failure of the ZP (zona pellucida) to rupture is believed to be associated with a decreased implantation rate in in vitro fertilization (IVF) or intra cytoplasmic sperm injection (ICSI). Objective: In this research, laser assisted hatching (LAH) was offered to patients with advanced maternal age to evaluate a possible benefit. Materials and Methods: Nine hundred thirty two cycles of IVF/ICSI in females were analyzed. Women included in this s...

  10. Laser-assisted new attachment procedure in private practice.

    Science.gov (United States)

    Harris, David M; Gregg, Robert H; McCarthy, Delwin K; Colby, Leigh E; Tilt, Lloyd V

    2004-01-01

    Three private dental practices conducted a retrospective analysis of patients receiving the laser-assisted new attachment procedure (LANAP). Retrospective results were compared to clinical trial data from the University of Texas Health Sciences Center in San Antonio (UTHSCSA) to determine if outcomes from a controlled clinical trial can be duplicated in private practice. Results also are compared with published results of other surgical and nonsurgical therapies for inflammatory periodontal disease.

  11. Leidenfrost phenomenon-assisted thermal desorption (LPTD) and its application to open ion sources at atmospheric pressure mass spectrometry.

    Science.gov (United States)

    Saha, Subhrakanti; Chen, Lee Chuin; Mandal, Mridul Kanti; Hiraoka, Kenzo

    2013-03-01

    This work describes the development and application of a new thermal desorption technique that makes use of the Leidenfrost phenomenon in open ion sources at atmospheric pressure for direct mass spectrometric detection of ultratrace levels of illicit, therapeutic, and stimulant drugs, toxicants, and peptides (molecular weight above 1 kDa) in their unaltered state from complex real world samples without or with minor sample pretreatment. A low temperature dielectric barrier discharge ion source was used throughout the experiments and the analytical figures of merit of this technique were investigated. Further, this desorption technique coupled with other ionization sources such as electrospray ionization (ESI) and dc corona discharge atmospheric pressure chemical ionization (APCI) in open atmosphere was also investigated. The use of the high-resolution 'Exactive Orbitrap' mass spectrometer provided unambiguous identification of trace levels of the targeted compounds from complex mixtures and background noise; the limits of detection for various small organic molecules and peptides treated with this technique were at the level of parts per trillion and 10(-9) M, respectively. The high sensitivity of the present technique is attributed to the spontaneous enrichment of analyte molecules during the slow evaporation of the solvent, as well as to the sequential desorption of molecules from complex mixtures based on their volatilities. This newly developed desorption technique is simple and fast, while molecular ions are observed as the major ions.

  12. Laser-induced electron transfer desorption/ionization of metal complexes on TiO2 films

    Science.gov (United States)

    Grechnikov, A. A.; Georgieva, V.; Borodkov, A. S.; Nikiforov, S. M.; Raicheva, Z.; Lazarov, J.; Donkov, N.

    2014-12-01

    Thin titanium dioxide (TiO2) films were studied as ion emitters for the laser-induced electron transfer desorption/ionization (LETDI) of metal complexes with organic reagents. The TiO2 films (350 nm thick) were deposited on the silicon substrates by e-beam evaporation of TiO2 powder. Copper complex with phthalocyanine, rhenium complex with thiocarbanilide and platinum complex with 8-quinolinethiol were studied as the test analytes. Reflectron time-of- flight mass spectrometer with the rotating ball interface was used for analysis. The analytes were applied on the surface of TiO2 film using an electrospray deposition. All tested compounds are detected as the radical molecular ions with no fragmentation. It is found, that TiO2 films are very stable and show good sensitivity in examined range of the analyte concentrations. The limits of detection of studied complexes were at the subfemtomole range, and the relative standard deviation was less than 10%.

  13. Effect of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry on identifing biomarkers of endometriosis

    Institute of Scientific and Technical Information of China (English)

    ZHANG Hong; FENG Jie; CHANG Xiao-hong; LI Zhong-xing; WU Xiao-yi; CUI Heng

    2009-01-01

    Background Endometriosis is a common gynecological disease. This study aimed to screen proteins that were expressed differently in patients with endometriosis versus normal controls using proteomic techniques, surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS).Methods Protein chip SELDI-TOF-MS combines the advantages of microarray and mass spectrometry, and can screen latent markers in sera of patients with endometriosis. Serum samples from patients and normal volunteers were analyzed by SELDI-TOF-MS.Results After comparing the serum protein spectra of 36 patients with 24 normal controls, 24 differently expressed potential biomarkers (P <0.01) were identified. Using Biomarker Pattern software, we established a tree model of the 60 serum protein spectra. When using the three biomarkers to classify the samples, the sensitivity for diagnosing endometriosis was 91.7%, specificity was 95.8%, and coincidence rate was 93.3%. Then we used serum samples from 12 patients and 8 normal controls to validate the tree model and report the sensitivity for diagnosing endometriosis was 91.7%, specificity was 75%, and coincidence rate was 85%.Conclusions SELDI-TOF-MS may be a useful tool in high-risk population screening for endometriosis. The identification and application of the biomarkers need to further study.

  14. Total microcystins analysis in water using laser diode thermal desorption-atmospheric pressure chemical ionization-tandem mass spectrometry.

    Science.gov (United States)

    Roy-Lachapelle, Audrey; Fayad, Paul B; Sinotte, Marc; Deblois, Christian; Sauvé, Sébastien

    2014-04-11

    A new approach for the analysis of the cyanobacterial microcystins (MCs) in environmental water matrices has been developed. It offers a cost efficient alternative method for the fast quantification of total MCs using mass spectrometry. This approach permits the quantification of total MCs concentrations without requiring any derivatization or the use of a suite of MCs standards. The oxidation product 2-methyl-3-methoxy-4-phenylbutyric acid (MMPB) was formed through a Lemieux oxidation and represented the total concentration of free and bound MCs in water samples. MMPB was analyzed using laser diode thermal desorption-atmospheric pressure chemical ionization coupled to tandem mass spectrometry (LDTD-APCI-MS/MS). LDTD is a robust and reliable sample introduction method with ultra-fast analysis time (0.999). Limits of detection and quantification were 0.2 and 0.9 μg L(-1), respectively. These values are comparable with the WHO (World Health Organization) guideline of 1 μg L(-1) for total microcystin-LR congener in drinking water. Accuracy and interday/intraday variation coefficients were below 15%. Matrix effect was determined with a recovery of 91%, showing no significant signal suppression. This work demonstrates the use of the LDTD-APCI-MS/MS interface for the screening, detection and quantification of total MCs in complex environmental matrices.

  15. Silica nanoparticles pre-spotted onto target plate for laser desorption/ionization mass spectrometry analyses of peptides.

    Science.gov (United States)

    Dupré, Mathieu; Cantel, Sonia; Durand, Jean-Olivier; Martinez, Jean; Enjalbal, Christine

    2012-09-05

    We report on the simple deposition of Stöber silica nanoparticles (SiO(2) NPs) on conventional MALDI target plate for high throughput laser desorption/ionization mass spectrometry (LDI-MS) analyses of peptide mixtures with sensitivity in the femtomolar range. This low-cost easily prepared material allowed straightforward LDI experiments by deposition of the studied samples directly onto a pre-spotted MALDI plate. This analytical strategy can be performed in any laboratory equipped with a MALDI-TOF instrument. All key benefits of organic matrix-free technologies were satisfied while maintaining a high level of detection performances (sensitivity and reproducibility/repeatability). In particular, sample preparation was simple and detection in the low mass range was not hampered by matrix ions. Imaging studies were undertaken to query sample dispersion into the inert SiO(2) NPs and to help into the search of the best experimental conditions producing homogeneous analyte distribution within the deposit. In contrast to commercial disposable LDI targets designed for single use and requiring an adaptor such as NALDI™, the proposed SiO(2) NPs pre-spotting on a MALDI target plate allowed very easily switching between MALDI and LDI experiments. They can be conducted either simultaneously (positions with an organic matrix or SiO(2) NPs) or in the row (support prepared in advance, stored and washed after use). The overall cost and versatility of the methodology made it very attractive to MALDI users in many domains (peptidomics, proteomics, metabolomics).

  16. Differentiation of Microbial Species and Strains in Coculture Biofilms by Multivariate Analysis of Laser Desorption Postionization Mass Spectra

    Energy Technology Data Exchange (ETDEWEB)

    University of Illinois at Chicago; Montana State University; Bhardwaj, Chhavi; Cui, Yang; Hofstetter, Theresa; Liu, Suet Yi; Bernstein, Hans C.; Carlson, Ross P.; Ahmed, Musahid; Hanley, Luke

    2013-04-01

    7.87 to 10.5 eV vacuum ultraviolet (VUV) photon energies were used in laser desorption postionization mass spectrometry (LDPI-MS) to analyze biofilms comprised of binary cultures of interacting microorganisms. The effect of photon energy was examined using both tunable synchrotron and laser sources of VUV radiation. Principal components analysis (PCA) was applied to the MS data to differentiate species in Escherichia coli-Saccharomyces cerevisiae coculture biofilms. PCA of LDPI-MS also differentiated individual E. coli strains in a biofilm comprised of two interacting gene deletion strains, even though these strains differed from the wild type K-12 strain by no more than four gene deletions each out of approximately 2000 genes. PCA treatment of 7.87 eV LDPI-MS data separated the E. coli strains into three distinct groups two ?pure? groups and a mixed region. Furthermore, the ?pure? regions of the E. coli cocultures showed greater variance by PCA when analyzed by 7.87 eV photon energies than by 10.5 eV radiation. Comparison of the 7.87 and 10.5 eV data is consistent with the expectation that the lower photon energy selects a subset of low ionization energy analytes while 10.5 eV is more inclusive, detecting a wider range of analytes. These two VUV photon energies therefore give different spreads via PCA and their respective use in LDPI-MS constitute an additional experimental parameter to differentiate strains and species.

  17. Characterization of aromaticity in analogues of titan's atmospheric aerosols with two-step laser desorption ionization mass spectrometry

    Science.gov (United States)

    Mahjoub, Ahmed; Schwell, Martin; Carrasco, Nathalie; Benilan, Yves; Cernogora, Guy; Szopa, Cyril; Gazeau, Marie-Claire

    2016-10-01

    The role of polycyclic aromatic hydrocarbons (PAH) and Nitrogen containing PAH (PANH) as intermediates of aerosol production in the atmosphere of Titan has been a subject of controversy for a long time. An analysis of the atmospheric emission band observed by the Visible and Infrared Mapping Spectrometer (VIMS) at 3.28 μm suggests the presence of neutral polycyclic aromatic species in the upper atmosphere of Titan. These molecules are seen as the counter part of negative and positive aromatics ions suspected by the Plasma Spectrometer onboard the Cassini spacecraft, but the low resolution of the instrument hinders any molecular speciation. In this work we investigate the specific aromatic content of Titan's atmospheric aerosols through laboratory simulations. We report here the selective detection of aromatic compounds in tholins, Titan's aerosol analogs, produced with a capacitively coupled plasma in a N2:CH4 95:5 gas mixture. For this purpose, Two-Step Laser Desorption Ionization Time-of-Flight Mass Spectrometry (L2DI-TOF-MS) technique is used to analyze the so produced analogs. This analytical technique is based on the ionization of molecules by Resonance Enhanced Multi-Photon Ionization (REMPI) using a λ=248 nm wavelength laser which is selective for aromatic species. This allows for the selective identification of compounds having at least one aromatic ring. Our experiments show that tholins contain a trace amount of small PAHs with one to three aromatic rings. Nitrogen containing PAHs (PANHs) are also detected as constituents of tholins. Molecules relevant to astrobiology are detected as is the case of the substituted DNA base adenine.

  18. Laser assisted Breit-Wheeler and Schwinger processes

    CERN Document Server

    Nousch, T; Seipt, D; Kämpfer, B; Titov, A I; Blaschke, D; Panferov, A D; Smolyansky, S A

    2016-01-01

    The assistance of an intense optical laser pulse on electron-positron pair production by the Breit-Wheeler and Schwinger processes in XFEL fields is analyzed. The impact of a laser beam on high-energy photon collisions with XFEL photons consists in a phase space redistribution of the pairs emerging in the Breit-Wheeler sub-process. We provide numerical examples of the differential cross section for parameters related to the European XFEL. Analogously, the Schwinger type pair production in pulsed fields with oscillating components referring to a superposition of optical laser and XFEL frequencies is evaluated. The residual phase space distribution of created pairs is sensitive to the pulse shape and may differ significantly from transiently achieved mode occupations.

  19. Laser-assisted treatment of patients with hemorrhagic diathesis

    Science.gov (United States)

    Neckel, Claus P.

    2000-03-01

    Today more and more patients with bleeding disorder come to our office for treatment. The number of patients with therapeutic anticoagulation is growing steadily. Discontinuation of this therapy can often be crucial. On the other hand are oralsurgical procedures extremely sensitive to bleeding due to the constant presents of saliva with its high fibrinolytic activity. The aim of this study was to evaluate the use of a surgical diode (wavelength 810 nm) laser as accessory tool in the treatment of patients with hemorrhagic diathesis. Enclosed in the study were 123 patients with: (1) Coumarintherapy, a Quick test ratio of 15 - 25%; (2) More than 300 mg of ASS/die; (3) Hemophilia, a factor activity under 35%; (4) Morbus Werlhof with less than 30000 thrombocytes. 179 Surgical procedures: (1) Tooth extraction 86%; (2) Apexectomy 3%; (3) Tumorexcision 9%; (4) Curettage and flapsurgery 1%; (5) Gingivectomy 1%. All procedures were laser-assisted with a diode laser emitting 810 nm. The glass fibers used were depending on the procedures either 200, 400, or 600 micron. No coagulating agents or tissue adhesives were used in addition. The postoperative outcome and complication rates were compared to substitution therapy and tissue adhesives. Laser-assisted treatment of these patients shows a high predictability and success rate leaving out side effects of drugs and human cryoprecipitates. Postoperative impairment is diminished.

  20. Online laser desorption-multiphoton postionization mass spectrometry of individual aerosol particles: molecular source indicators for particles emitted from different traffic-related and wood combustion sources.

    Science.gov (United States)

    Bente, Matthias; Sklorz, Martin; Streibel, Thorsten; Zimmermann, Ralf

    2008-12-01

    Direct inlet aerosol mass spectrometry plays an increasingly important role in applied and fundamental aerosol and nanoparticle research. Laser desorption/ionization (LDI) based techniques for single particle time-of-flight mass spectrometry (LDI-SP-TOFMS) are a promising approach in the chemical analysis of single aerosol particles, especially for the detection of inorganic species and distinction of particle classes. However, until now the detection of molecular organic compounds on a single particle basis has been difficult due to the high laser power densities which are required for the LDI process as well as due to the inherent matrix effects associated with this ionization technique. By the application of a two-step approach, where an IR desorption laser pulse is applied to perform a gentle desorption of organic material from the single particle surface and a second UV-laser performs the soft ionization of the desorbed species, this drawback of laser based single particles mass spectrometry can be overcome. The postionization of the desorbed molecules has been accomplished in this work by resonance enhanced multiphoton ionization (REMPI) using a KrF excimer laser (248 nm). REMPI allows an almost fragmentation free trace analysis of polycyclic aromatic hydrocarbons (PAHs) and their derivatives from individual single particles (laser desorption-REMPI postionization-single particle-time-of-flight mass spectrometry or LD-REMPI-SP-TOFMS). Crucial system parameters of the home-built aerosol mass spectrometer such as the power densities and the relative timing of both lasers were optimized with respect to the detectability of particle source specific organic signatures using well characterized standard particles. In a second step, the LD-REMPI-SP-TOFMS system was applied to analyze different real world aerosols (spruce wood combustion, gasoline car exhaust, beech wood combustion, and diesel car exhaust). It was possible to distinguish the particles from different

  1. Laser-assisted advanced assembly for MEMS fabrication

    Science.gov (United States)

    Atanasov, Yuriy Andreev

    Micro Electro-Mechanical Systems (MEMS) are currently fabricated using methods originally designed for manufacturing semiconductor devices, using minimum if any assembly at all. The inherited limitations of this approach narrow the materials that can be employed and reduce the design complexity, imposing limitations on MEMS functionality. The proposed Laser-Assisted Advanced Assembly (LA3) method solves these problems by first fabricating components followed by assembly of a MEMS device. Components are micro-machined using a laser or by photolithography followed by wet/dry etching out of any material available in a thin sheet form. A wide range of materials can be utilized, including biocompatible metals, ceramics, polymers, composites, semiconductors, and materials with special properties such as memory shape alloys, thermoelectric, ferromagnetic, piezoelectric, and more. The approach proposed allows enhancing the structural and mechanical properties of the starting materials through heat treatment, tribological coatings, surface modifications, bio-functionalization, and more, a limited, even unavailable possibility with existing methods. Components are transferred to the substrate for assembly using the thermo-mechanical Selective Laser Assisted Die Transfer (tmSLADT) mechanism for microchips assembly, already demonstrated by our team. Therefore, the mechanical and electronic part of the MEMS can be fabricated using the same equipment/method. The viability of the Laser-Assisted Advanced Assembly technique for MEMS is demonstrated by fabricating magnetic switches for embedding in a conductive carbon-fiber metamaterial for use in an Electromagnetic-Responsive Mobile Cyber-Physical System (E-RMCPS), which is expected to improve the wireless communication system efficiency within a battery-powered device.

  2. Fundamentals of laser-assisted fabrication of inorganic and organic films

    DEFF Research Database (Denmark)

    Schou, Jørgen

    2008-01-01

    The standard method for producing films by laser-assisted methods, Pulsed Laser Deposition (PLD) will be reviewed. The films considered are usually inorganic films, but also films of organic materials have been produced. Also the deposition of organic films by MAPLE (Matrix Assisted Pulsed Laser ...

  3. MATRIX ASSISTED LASER DESORPTION/IONIZATION TIME OF FLIGHT MASS SPECTROMETRY BASED ANALYSIS OF GIARDIA LAMBLIA

    Science.gov (United States)

    Giardia lamblia is a zoonotic protozoan parasite that is a leading cause of drinking water related gastro-intestinal disease outbreaks worldwide. Due to the genotypic complexity and high prevalence of this parasite in the environment, numerous research studies are being done to ...

  4. Probing of Metabolites in Finely Powdered Plant Material by Direct Laser Desorption Ionization Mass Spectrometry

    Science.gov (United States)

    Musharraf, Syed Ghulam; Ali, Arslan; Choudhary, M. Iqbal; Atta-ur-Rahman

    2014-04-01

    Natural products continue to serve as an important source of novel drugs since the beginning of human history. High-throughput techniques, such as MALDI-MS, can be techniques of choice for the rapid screening of natural products in plant materials. We present here a fast and reproducible matrix-free approach for the direct detection of UV active metabolites in plant materials without any prior sample preparation. The plant material is mechanically ground to a fine powder and then sieved through different mesh sizes. The collected plant material is dispersed using 1 μL solvent on a target plate is directly exposed to Nd:YAG 335 nm laser. The strategy was optimized for the analysis of plant metabolites after study of the different factors affecting the reproducibility and effectiveness of the analysis, including particle sizes effects, types of solvents used to disperse the sample, and the part of the plant analyzed. Moreover, several plant species, known for different classes of metabolites, were screened to establish the generality of the approach. The developed approach was validated by the characterization of withaferin A and nicotine in the leaves of Withania somnifera and Nicotiana tabacum, respectively, through comparison of its MS/MS data with the standard compound. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) techniques were used for the tissue imaging purposes. This approach can be used to directly probe small molecules in plant materials as well as in herbal and pharmaceutical formulations for fingerprinting development.

  5. Matrix-Assisted Pulsed Laser Evaporation of polythiophene films

    Energy Technology Data Exchange (ETDEWEB)

    Bloisi, F. [CNR-INFM Coherentia, Napoli, Dip. Scienze Fisiche, Univ. Napoli ' Federico II' , P.le V.Tecchio, 80, 80125 Naples (Italy)], E-mail: bloisi@na.infn.it; Cassinese, A.; Papa, R.; Vicari, L. [CNR-INFM Coherentia, Napoli, Dip. Scienze Fisiche, Univ. Napoli ' Federico II' , P.le V.Tecchio, 80, 80125 Naples (Italy); Califano, V. [Dip. Scienze Fisiche, Univ. Napoli ' Federico II' , P.le V.Tecchio, 80, 80125 Naples (Italy)

    2008-02-15

    Organic poly-conjugated systems have recently attracted great interest as semi-conducting materials and, among poly-conjugated systems, substituted polythiophenes have given relevant results in PVs applications. The high conductivity required is affected by both the polymer conjugation length and the chain packing. Thus, highly region-regular polymers must be used and deposited as thin films with some technique which favours orientation and crystallization of the polymer chains. A deposition technique often used for its flexibility and high control over film characteristics is Pulsed Laser Deposition (PLD). In PLD, largely applied for inorganic thin film deposition, the material is ablated from a solid target by a focused pulsed laser beam and is deposited on the substrate placed at a small distance. Although some addition polymers have been successfully deposited the deposition seems to proceed via a 'depolymerization-monomer ablation-repolymerization' mechanism, this is clearly not possible in general for organic molecules and condensation polymers. On the contrary MAPLE (Matrix-Assisted Pulsed Laser Evaporation) is a recently developed PLD based thin film deposition technique, particularly well suited for organic/polymer thin film deposition. Up to now MAPLE depositions have been carried out mainly by means of modified PLD systems, using excimer lasers operating in UV, but use of less energetic radiations can minimize the photochemical decomposition of the polymer molecules. We have used a deposition system explicitly designed for MAPLE technique connected to a Q-switched Ng:YAG pulsed laser which can be operated at different wavelength ranging from IR to UV in order to evaluate the effect of the choice of laser radiation on the deposition of POOPT thin films. From DRIFT-IR spectroscopy, all deposited films showed structural order; it was determined that the better wavelength for POOPT deposition is 532 nm. With this value of the laser wavelength the

  6. An investigation on co-axial water-jet assisted fiber laser cutting of metal sheets

    Science.gov (United States)

    Madhukar, Yuvraj K.; Mullick, Suvradip; Nath, Ashish K.

    2016-02-01

    Water assisted laser cutting has received significant attention in recent times with assurance of many advantages than conventional gas assisted laser cutting. A comparative study between co-axial water-jet and gas-jet assisted laser cutting of thin sheets of mild steel (MS) and titanium (Ti) by fiber laser is presented. Fiber laser (1.07 μm wavelength) was utilised because of its low absorption in water. The cut quality was evaluated in terms of average kerf, projected dross height, heat affected zone (HAZ) and cut surface roughness. It was observed that a broad range process parameter could produce consistent cut quality in MS. However, oxygen assisted cutting could produce better quality only with optimised parameters at high laser power and high cutting speed. In Ti cutting the water-jet assisted laser cutting performed better over the entire range of process parameters compared with gas assisted cutting. The specific energy, defined as the amount of laser energy required to remove unit volume of material was found more in case of water-jet assisted laser cutting process. It is mainly due to various losses associated with water assisted laser processing such as absorption of laser energy in water and scattering at the interaction zone.

  7. Large-Scale Metabolite Analysis of Standards and Human Serum by Laser Desorption Ionization Mass Spectrometry from Silicon Nanopost Arrays.

    Science.gov (United States)

    Korte, Andrew R; Stopka, Sylwia A; Morris, Nicholas; Razunguzwa, Trust; Vertes, Akos

    2016-09-20

    The unique challenges presented by metabolomics have driven the development of new mass spectrometry (MS)-based techniques for small molecule analysis. We have previously demonstrated silicon nanopost arrays (NAPA) to be an effective substrate for laser desorption ionization (LDI) of small molecules for MS. However, the utility of NAPA-LDI-MS for a wide range of metabolite classes has not been investigated. Here we apply NAPA-LDI-MS to the large-scale acquisition of high-resolution mass spectra and tandem mass spectra from a collection of metabolite standards covering a range of compound classes including amino acids, nucleotides, carbohydrates, xenobiotics, lipids, and other classes. In untargeted analysis of metabolite standard mixtures, detection was achieved for 374 compounds and useful MS/MS spectra were obtained for 287 compounds, without individual optimization of ionization or fragmentation conditions. Metabolite detection was evaluated in the context of 31 metabolic pathways, and NAPA-LDI-MS was found to provide detection for 63% of investigated pathway metabolites. Individual, targeted analysis of the 20 common amino acids provided detection of 100% of the investigated compounds, demonstrating that improved coverage is possible through optimization and targeting of individual analytes or analyte classes. In direct analysis of aqueous and organic extracts from human serum samples, spectral features were assigned to a total of 108 small metabolites and lipids. Glucose and amino acids were quantitated within their physiological concentration ranges. The broad coverage demonstrated by this large-scale screening experiment opens the door for use of NAPA-LDI-MS in numerous metabolite analysis applications.

  8. Kinetics of Laser-Assisted Carbon Nanotube Growth

    CERN Document Server

    van de Burgt, Yoeri; Mandamparambil, Rajesh

    2014-01-01

    Laser-assisted chemical vapour deposition (CVD) growth is an attractive mask-less process for growing locally aligned carbon nanotubes (CNTs) in selected places on temperature sensitive substrates. The nature of the localized process results in fast carbon nanotube growth with high experimental throughput. Here, we report on detailed investigation of growth kinetics related to physical and chemical process characteristics. Specifically, the growth kinetics is investigated by monitoring the dynamical changes of reflected laser beam intensity during growth. Benefiting from the fast growth and high experimental throughput, we investigate a wide range of experimental conditions and propose several growth regimes. Rate-limiting steps are determined using rate equations linked to the proposed growth regimes, which are further characterized by Raman spectroscopy and Scanning Electron Microscopy (SEM), therefore directly linking growth regimes to the structural quality of the CNTs. Activation energies for the differe...

  9. Laser-Assisted Elastic Electron Scattering from Argon

    Institute of Scientific and Technical Information of China (English)

    HU Qiu-Bo; SUN Jin-Feng

    2009-01-01

    The second Born approximation (SBA) theory is applied to the study of electron-atom scattering in the presence of a CO2 laser field. The absolute differential cross sections of e-At scattering are calculated with multiphoton exchange in two special scattering geometries G1 (for small-angle scattering) and G2. For geometry G1, compared with the results of two different model potentials for electron elastic scattering by atoms, it is found that electronatom polarization potential plays an important role in laser-assisted electron-atom scattering. Some calculational results in geometries G2 are given. Our results are found to be better than other theoretical results as compared with the experimental data in geometries G1 and G2.

  10. Decay-Assisted Laser Spectroscopy of Neutron-Deficient Francium

    CERN Document Server

    Lynch, K M; Bissell, M L; Budincevic, I; Cocolios, T E; De Groote, R P; De Schepper, S; Fedosseev, V N; Flanagan, K T; Franchoo, S; Garcia Ruiz, R F; Heylen, H; Marsh, B A; Neyens, G; Procter, T J; Rossel, R E; Rothe, S; Strashnov, I; Stroke, H H; Wendt, K D A

    2014-01-01

    This paper reports on the hyperfine-structure and radioactive-decay studies of the neutron-deficient francium isotopes $^{202-206}$Fr performed with the Collinear Resonance Ionization Spectroscopy (CRIS) experiment at the ISOLDE facility, CERN. The high resolution innate to collinear laser spectroscopy is combined with the high efficiency of ion detection to provide a highly-sensitive technique to probe the hyperfine structure of exotic isotopes. The technique of decay-assisted laser spectroscopy is presented, whereby the isomeric ion beam is deflected to a decay spectroscopy station for alpha-decay tagging of the hyperfine components. Here, we present the first hyperfine-structure measurements of the neutron-deficient francium isotopes $^{202-206}$Fr, in addition to the identification of the low-lying states of $^{202,204}$Fr performed at the CRIS experiment.

  11. Bioceramic 3D Implants Produced by Laser Assisted Additive Manufacturing

    Science.gov (United States)

    Lusquiños, Fernando; del Val, Jesús; Arias-González, Felipe; Comesaña, Rafael; Quintero, Félix; Riveiro, Antonio; Boutinguiza, Mohamed; Jones, Julian R.; Hill, Robert G.; Pou, Juan

    Cranial defect restoration requires a suitable implant capable to fulfill protective and aesthetic functions, such as polymeric and metallic implants. Nevertheless, the former materials cannot provide osteointegration of the implant within the host bone nor implant resorption, which is also required in pediatricorthopedics for normal patient growth. Resorbable and osteoconductivebioceramics are employed, such as silicate bioactive glasses. Nevertheless, manufacturing based on conventional casting in graphite moulds is not effective for warped shape implants suitable for patient tailored treatments. In this work, we analyze the application of rapid prototyping based on laser cladding to manufacture bioactive glass implants for low load bearing bone restoration. This laser-assisted additive technique is capable to produce three-dimensional geometries tailored to patient, with reduced fabrication time and implant composition modification. The obtained samples were characterized; the relationships between the processing conditions and the measured features were studied, in addition to the biological behavior analysis.

  12. Lipase biofilm deposited by Matrix Assisted Pulsed Laser Evaporation technique

    Science.gov (United States)

    Aronne, Antonio; Bloisi, Francesco; Calabria, Raffaela; Califano, Valeria; Depero, Laura E.; Fanelli, Esther; Federici, Stefania; Massoli, Patrizio; Vicari, Luciano R. M.

    2015-05-01

    Lipase is an enzyme that finds application in biodiesel production and for detection of esters and triglycerides in biosensors. Matrix Assisted Pulsed Laser Evaporation (MAPLE), a technique derived from Pulsed Laser Deposition (PLD) for deposition of undamaged biomolecules or polymers, is characterized by the use of a frozen target obtained from a solution/suspension of the guest material (to be deposited) in a volatile matrix (solvent). The presence of the solvent avoids or at least reduces the potential damage of guest molecules by laser radiation but only the guest material reaches the substrate in an essentially solvent-free deposition. MAPLE can be used for enzymes immobilization, essential for industrial application, allowing the development of continuous processes, an easier separation of products, the reuse of the catalyst and, in some cases, enhancing enzyme properties (pH, temperature stability, etc.) and catalytic activity in non-aqueous media. Here we show that MAPLE technique can be used to deposit undamaged lipase and that the complex structure (due to droplets generated during extraction from target) of the deposited material can be controlled by changing the laser beam fluence.

  13. Ex vivo laser lipolysis assisted with radially diffusing optical applicator

    Science.gov (United States)

    Hwang, Jieun; Hau, Nguyen Trung; Park, Sung Yeon; Rhee, Yun-Hee; Ahn, Jin-Chul; Kang, Hyun Wook

    2016-05-01

    Laser-assisted lipolysis has been implemented to reduce body fat in light of thermal interactions with adipose tissue. However, using a flat fiber with high irradiance often needs rapid cannula movements and even undesirable thermal injury due to direct tissue contact. The aim of the current study was to explore the feasibility of a radially diffusing optical applicator to liquefy the adipose tissue for effective laser lipolysis. The proposed diffuser was evaluated with a flat fiber in terms of temperature elevation and tissue liquefaction after laser lipolysis with a 980-nm wavelength. Given the same power (20 W), the diffusing applicator generated a 30% slower temperature increase with a 25% lower maximum temperature (84±3.2°C in 1 min ptissue, compared with the flat fiber. Under the equivalent temperature development, the diffuser induced up to fivefold larger area of the adipose liquefaction due to radial light emission than the flat fiber. Ex vivo tissue tests for 5-min irradiation demonstrated that the diffuser (1.24±0.15 g) liquefied 66% more adipose tissue than the flat fiber (0.75±0.05 g). The proposed diffusing applicator can be a feasible therapeutic device for laser lipolysis due to low temperature development and wide coverage of thermal treatment.

  14. Laser-assisted cleaning: Dominant role of surface

    Indian Academy of Sciences (India)

    J Padma Nilaya; D J Biswas

    2010-12-01

    Pulsed laser-assisted removal of particulates from substrates has decided advantages over the conventional methods of cleaning. Experiments conducted with loose contamination on metal and transparent dielectric surfaces proved conclusively the dominant role played by the absorption of the incident radiation by the surface towards the generation of the cleaning force as against the absorption in the particulates alone. Further, the presence of transparent/semi-transparent particulates on a metal surface was found to result in an increased absorption of the incident radiation by the substrate. This effect, identified as field-enhanced surface absorption was found to increase with reduction in the average particulate size.

  15. Matrix-assisted pulsed laser evaporation of chemoselective polymers

    Science.gov (United States)

    Palla-Papavlu, Alexandra; Dinca, Valentina; Dinescu, Maria; di Pietrantonio, Fabio; Cannatà, Domenico; Benetti, Massimiliano; Verona, Enrico

    2011-11-01

    In this work, matrix-assisted pulsed laser evaporation was applied to achieve gentle deposition of polymer thin films onto surface acoustic wave resonators. Polyepichlorhydrin, polyisobutylene and polyethylenimine were deposited both onto rigid substrates e.g. Si wafers as well as surface acoustic wave devices using a Nd-YAG laser (266 nm, 355 nm, 10 Hz repetition rate). Morphological investigations (atomic force microscopy and optical microscopy) reveal continuous deposited polymer thin films, and in the case of polyethylenimine a very low surface roughness of 1.2 nm (measured on a 40×40 μm2 area). It was found that only for a narrow range of laser fluences (i.e. 0.1-0.3 J/cm2 in the case of polyisobutylene) the chemical structure of the deposited polymer thin layers resembles to the native polymer. In addition, in the case of polyisobutylene it was shown that the irradiation at 355-nm wavelength produces deviations in the chemical structure of the deposited polymer, as compared to its bulk structure. Following the morphological and structural characterization, only a set of well established conditions was used for polymer deposition on the sensor structures. The surface acoustic wave resonators have been tested using the Network Analyzer before and after polymer deposition. The polymer coated surface acoustic wave resonator responses have been measured upon exposure to various concentrations of dimethylmethylphosphonate analyte. All sensors coated with different polymer layers (polyethylenimine, polyisobutylene, and polyepichlorhydrin) show a clear response to the dimethylmethylphosphonate vapor. The strongest signal is obtained for polyisobutylene, followed by polyethylenimine and polyepichlorhydrin. The results obtained indicate that matrix-assisted pulsed laser evaporation is potentially useful for the fabrication of polymer thin films to be used in applications including microsensor industry.

  16. Size-effects on energy relaxation and excited-species desorption in krypton clusters: Fluorescence lifetime measurements with 10 eV laser excitation

    Science.gov (United States)

    Kanaev, A. V.; Museur, L.; Castex, M. C.

    1997-09-01

    Fluorescence lifetime measurements of KrN clusters (N¯=2-2000) have been carried out using intense 10 eV laser excitation near 3P2 metastable atomic energy level. Two principal groups of electronically excited dimers Kr2* have been found in desorption: dimers, loosely bound near the (3P2+1S0) dissociation limit, ejected from cooled clusters and dimers undergoing vibrational relaxation from hot clusters. The desorption is principally terminated when N¯⩾50 at./cluster. The relaxation kinetics seems to converge to the properties of a solid state for 102⩽N¯⩽103 at./cluster. A variation of the Kr2*(1u/0u-) radiative lifetime, from 264 ns (in gas phase) to 440 ns (N¯=102), has been found. An equilibrium cluster temperature of 57 K has been calculated from this τ(N) dependence.

  17. Application of microwave-assisted desorption/headspace solid-phase microextraction as pretreatment step in the gas chromatographic determination of 1-naphthylamine in silica gel adsorbent.

    Science.gov (United States)

    Yan, Cheing-Tong; Jen, Jen-Fon; Shih, Tung-Sheng

    2007-03-30

    Pretreatment of silica gel sample containing 1-naphthylamine by microwave-assisted desorption (MAD) coupled to in situ headspace solid phase microextraction (HS-SPME) has been investigated as a possible alternative to conventional methods prior to gas chromatographic (GC) analysis. The 1-naphthylamine desorbs from silica gel to headspace under microwave irradiation, and directly absorbs onto a SPME fiber located in a controlled-temperature headspace area. After being collected on the SPME fiber, and desorbed in the GC injection port, 1-naphthylamine is analyzed by GC-FID. Parameters that influence the extraction efficiency of the MAD/HS-SPME, such as the extraction media and its pH, the microwave irradiation power and irradiation time as well as desorption conditions of the GC injector, have been investigated. Experimental results indicate that the extraction of a 150mg silica gel sample by using 0.8ml of 1.0M NaOH solution and a PDMS/DVB fiber under high-powered irradiation (477W) for 5min maximizes the extraction efficiency. Desorption of 1-naphthylamine from the SPME fiber in GC injector is optimal at 250 degrees C held for 3min. The detection limit of method is 8.30ng. The detected quantity of 1-naphthylamine obtained by the proposed method is 33.3 times of that obtained by the conventional solvent extraction method for the silica gel sample containing 100ng of 1-naphthylamine. It provides a simple, fast, sensitive and organic-solvent-free pretreatment procedure prior to the analysis of 1-naphthylamine collected on a silica gel adsorbent.

  18. LIFT: analysis of performance in a laser assisted adaptive optics

    Science.gov (United States)

    Plantet, Cedric; Meimon, Serge; Conan, Jean-Marc; Neichel, Benoît; Fusco, Thierry

    2014-08-01

    Laser assisted adaptive optics systems rely on Laser Guide Star (LGS) Wave-Front Sensors (WFS) for high order aberration measurements, and rely on Natural Guide Stars (NGS) WFS to complement the measurements on low orders such as tip-tilt and focus. The sky-coverage of the whole system is therefore related to the limiting magnitude of the NGS WFS. We have recently proposed LIFT, a novel phase retrieval WFS technique, that allows a 1 magnitude gain over the usually used 2×2 Shack-Hartmann WFS. After an in-lab validation, LIFT's concept has been demonstrated on sky in open loop on GeMS (the Gemini Multiconjugate adaptive optics System at Gemini South). To complete its validation, LIFT now needs to be operated in closed loop in a laser assisted adaptive optics system. The present work gives a detailed analysis of LIFT's behavior in presence of high order residuals and how to limit aliasing effects on the tip/tilt/focus estimation. Also, we study the high orders' impact on noise propagation. For this purpose, we simulate a multiconjugate adaptive optics loop representative of a GeMS-like 5 LGS configuration. The residual high orders are derived from a Fourier based simulation. We demonstrate that LIFT keeps a high performance gain over the Shack-Hartmann 2×2 whatever the turbulence conditions. Finally, we show the first simulation of a closed loop with LIFT estimating turbulent tip/tilt and focus residuals that could be induced by sodium layer's altitude variations.

  19. Minimally invasive laser-assisted treatment of arytenoid chondritis

    Science.gov (United States)

    Sullins, Kenneth E.

    2001-05-01

    Five adult performance horses presented with the complaints of exercise intolerance and/or upper airway noise. Endoscopy revealed reduced arytenoid movement, mucosal defects with protruding granulation masses and occasional perilaryngeal swelling. While standing, a 1-cm stab incision was created, and a 5-mm trocar with sleeve was inserted through the cricothyroid membrane using nasopharyngeal endoscopic guidance. A fiber guide containing a 600-micron diode free-beam laser fiber was inserted, and the masses were ablated to the cartilage surface. Lesions determined to be limited to the cartilage substance were further ablated using the laser. Lesions determined to extend through the cartilage were manually debrided using an angled curette. The wounds were left unsutured after placing 2-3 ml of antiseptic ointment subcutaneously. Reexamination revealed resolution of the lesions in all horses, and arytenoid mobility was present. Lesions affecting the corniculate process (two) resulted in noticeable atrophy. The author concludes that laser-assisted debridement of septic tracts in the arytenoid cartilages that have not become completely deformed by the process is a reasonable procedure to restore athletic function.

  20. Resident surgeon efficiency in femtosecond laser-assisted cataract surgery

    Science.gov (United States)

    Pittner, Andrew C; Sullivan, Brian R

    2017-01-01

    Purpose Comparison of resident surgeon performance efficiencies in femtosecond laser-assisted cataract surgery (FLACS) versus conventional phacoemulsification. Patients and methods A retrospective cohort study was conducted on consecutive patients undergoing phacoemulsification cataract surgery performed by senior ophthalmology residents under the supervision of 1 attending physician during a 9-month period in a large Veterans Affairs medical center. Medical records were reviewed for demographic information, preoperative nucleus grade, femtosecond laser pretreatment, operative procedure times, total operating room times, and surgical complications. Review of digital video records provided quantitative interval measurements of core steps of the procedures, including completion of incisions, anterior capsulotomy, nucleus removal, cortical removal, and intraocular lens implantation. Results Total room time, operation time, and corneal incision completion time were found to be significantly longer in the femtosecond laser group versus the traditional phacoemulsification group (each Pcataract surgery is generally less efficient when trainees have more experience with traditional phacoemulsification. FLACS was found to have a significant advantage in completion of capsulotomy, but subsequent surgical steps were not shorter or longer. Resident learning curve for the FLACS technology may partially explain the disparities of performance. Educators should be cognizant of a potential for lower procedural efficiency when introducing FLACS into resident training. PMID:28203055

  1. In situ probing of cholesterol in astrocytes at the single-cell level using laser desorption ionization mass spectrometric imaging with colloidal silver.

    Science.gov (United States)

    Perdian, D C; Cha, Sangwon; Oh, Jisun; Sakaguchi, Donald S; Yeung, Edward S; Lee, Young Jin

    2010-04-30

    Mass spectrometric imaging has been utilized to localize individual astrocytes and to obtain cholesterol populations at the single-cell level in laser desorption ionization (LDI) with colloidal silver. The silver ion adduct of membrane-bound cholesterol was monitored to detect individual cells. Good correlation between mass spectrometric and optical images at different cell densities indicates the ability to perform single-cell studies of cholesterol abundance. The feasibility of quantification is confirmed by the agreement between the LDI-MS ion signals and the results from a traditional enzymatic fluorometric assay. We propose that this approach could be an effective tool to study chemical populations at the cellular level.

  2. Parametric modeling and optimization of laser scanning parameters during laser assisted machining of Inconel 718

    Science.gov (United States)

    Venkatesan, K.; Ramanujam, R.; Kuppan, P.

    2016-04-01

    This paper presents a parametric effect, microstructure, micro-hardness and optimization of laser scanning parameters (LSP) on heating experiments during laser assisted machining of Inconel 718 alloy. The laser source used for experiments is a continuous wave Nd:YAG laser with maximum power of 2 kW. The experimental parameters in the present study are cutting speed in the range of 50-100 m/min, feed rate of 0.05-0.1 mm/rev, laser power of 1.25-1.75 kW and approach angle of 60-90°of laser beam axis to tool. The plan of experiments are based on central composite rotatable design L31 (43) orthogonal array. The surface temperature is measured via on-line measurement using infrared pyrometer. Parametric significance on surface temperature is analysed using response surface methodology (RSM), analysis of variance (ANOVA) and 3D surface graphs. The structural change of the material surface is observed using optical microscope and quantitative measurement of heat affected depth that are analysed by Vicker's hardness test. The results indicate that the laser power and approach angle are the most significant parameters to affect the surface temperature. The optimum ranges of laser power and approach angle was identified as 1.25-1.5 kW and 60-65° using overlaid contour plot. The developed second order regression model is found to be in good agreement with experimental values with R2 values of 0.96 and 0.94 respectively for surface temperature and heat affected depth.

  3. Laser assisted die bending: a new application of high power diode lasers

    Science.gov (United States)

    Schuöcker, D.; Schumi, T.; Spitzer, O.; Bammer, F.; Schuöcker, G.; Sperrer, G.

    2015-02-01

    Nowadays high power lasers are mainly used for cutting of sheet metals, for welding, hardening and rapid prototyping. In the forming of sheet metals as bending or deep drawing lasers are not used. Nevertheless a few years ago a new application of high power lasers has been invented, where bending of materials that break at room temperature becomes possible by heating them along the bending edge with high power lasers thus allowing their treatment without cracks and rupture. For this purpose a large number of diode lasers are arranged in the bottom tool of a bending machine (a V-shaped die) which heat up the initially flat sheet metal during the bending process what is performed by pressing it into the die with a knife shaped upper tool where due to the laser heating the material is softened and thus cracks are avoided. For the technical realization of the new process of laser assisted die bending, modules equipped with numerous laser diodes and a total beam power of 2,5 kW are used. The light emitted by these modules enters a tool with a length of 15cm and is deflected towards the workpiece. By using ten of these modules with adjacent dies and by integrating those in a bending press a bending edge of sheet metals with a length of 1500mm can be realized. Such a bending press with laser assistance also needs energization with a power of practically 50kW, a respective water flow, a heat exchanger system and also a control for all functions of this system. Special measures have also been developed to avoid radiating of those tools that are not covered by a workpiece in the case of bending edges shorter than the full length of the bending tools whereas individual short circuiting of diode modules can be performed. Specific measures to ensure a safe operation without any harm to the operational person have been realized. Exploitation of the bending process has been carried out for titanium, where material thicknesses up to 3mm have been bent successfully.

  4. Posterior Corneal Surface Stability after Femtosecond Laser-Assisted Keratomileusis

    Directory of Open Access Journals (Sweden)

    Carlo Cagini

    2015-01-01

    Full Text Available The purpose of this study was to evaluate posterior corneal surface variation after femtosecond laser-assisted keratomileusis in patients with myopia and myopic astigmatism. Patients were evaluated by corneal tomography preoperatively and at 1, 6, and 12 months. We analyzed changes in the posterior corneal curvature, posterior corneal elevation, and anterior chamber depth. Moreover, we explored correlation between corneal ablation depth, residual corneal thickness, percentage of ablated corneal tissue, and preoperative corneal thickness. During follow-up, the posterior corneal surface did not have a significant forward corneal shift: no significant linear relationships emerged between the anterior displacement of the posterior corneal surface and corneal ablation depth, residual corneal thickness, or percentage of ablated corneal tissue.

  5. Chromophore-assisted laser inactivation in neural development

    Institute of Scientific and Technical Information of China (English)

    Wei Li; Nico Stuurman; Guangshuo Ou

    2012-01-01

    Chromophore-assisted laser inactivation (CALI) is a technique that uses photochemically-generated reactive oxygen species to acutely inactivate target proteins in living cells.Neural development includes highly dynamic cellular processes such as asymmetric cell division,migration,axon and dendrite outgrowth and synaptogenesis.Although many key molecules of neural development have been identified since the past decades,their spatiotemporal contributions to these cellular events are not well understood.CALI provides an appealing tool for elucidating the precise functions of these molecules during neural development.In this review,we summarize the principles of CALI,a recent microscopic setup to perform CALI experiments,and the application of CALI to the study of growth-cone motility and neuroblast asymmetric division.

  6. Using optical pyrometer in gas assisted laser cutting of steel sheets

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    @@ The use of optical methods of nondestructive testing can permit the real time control of gas assisted laser cutting, which is especially important in cutting metal sheets of large thickness and in other cases when the specified laser power is limited by laser operation modes[1,2].

  7. Using optical pyrometer in gas assisted laser cutting of steel sheets

    Institute of Scientific and Technical Information of China (English)

    Grishaev; R.; V.; Dubrov; V.; D.; Dubrovin; N.; G.; Zavalov; Yu.; N.

    2005-01-01

    The use of optical methods of nondestructive testing can permit the real time control of gas assisted laser cutting, which is especially important in cutting metal sheets of large thickness and in other cases when the specified laser power is limited by laser operation modes[1,2].……

  8. Frequency-comb-assisted broadband precision spectroscopy with cascaded diode lasers

    DEFF Research Database (Denmark)

    Liu, Junqiu; Brasch, Victor; Pfeiffer, Martin H. P.;

    2016-01-01

    Frequency-comb-assisted diode laser spectroscopy, employing both the accuracy of an optical frequency comb and the broad wavelength tuning range of a tunable diode laser, has been widely used in many applications. In this Letter, we present a novel method using cascaded frequency agile diode lasers...

  9. Surgical adhesives for laser-assisted wound closure

    Science.gov (United States)

    Hodges, Diane E.; McNally-Heintzelman, Karen M.; Welch, Ashley J.

    2001-10-01

    Solid protein solder-doped polymer membranes were developed for laser-assisted tissue repair. Biodegradable polymer membranes of controlled porosity were fabricated with poly(L-lactic-co-glycolic acid) (PLGA), poly(ethylene glycol) (PEG), and salt particles, using a solvent-casting and particulate-leaching technique. The membranes provided a porous scaffold that readily absorbed the traditional protein solder composed of serum albumin, indocyanine green dye, and de-ionized water. In vitro investigations were conducted to assess the influence of various processing parameters on the strength of tissue repairs formed using the new membranes. These parameters included PLGA copolymer and PLGA/PEG blend ratios, membrane pore size, initial albumin weight fraction, and laser irradiance used to denature the solder. Altering the PLGA copolymer ratio had little effect on repair strength, however such variations are known to influence the degradation rate of the membranes. The repair strength increased with increased membrane pore size and bovine serum albumin concentration. The addition of PEG during the membrane casting stage increased the flexibility of the membranes but not necessarily the repair strength. Typically, the repair strength increased with increasing irradiance from 12 to 18 W/cm2. The new solder-doped polymer membranes provided all of the benefits associated with solid protein solders including high repair strength and improved edge coaptation. In addition, the flexible, moldable nature of the new membranes offers the capability of tailoring the membranes to a wide range of clinically relevant geometries.

  10. Dacriocistorrinostomia transcanalicular com laser diodo: resultados preliminares Transcanalicular laser-assisted dacryocystorhinostomy: initial results

    Directory of Open Access Journals (Sweden)

    Ricardo Tomoyoshi Kanecadan

    2006-10-01

    Full Text Available OBJETIVO: Descrever a técnica e os resultados iniciais da dacriocistorrinostomia com laser diodo, realizada pela via transcanalicular. MÉTODOS: Dez pacientes com diagnóstico de obstrução lacrimal baixa foram submetidos à dacriocistorrinostomia com laser diodo, realizada pela via transcanalicular. A via lacrimal foi entubada com silicone, onde deveria permanecer por 6 meses. RESULTADOS: Todas as dez cirurgias foram realizadas sem intercorrências. Um paciente apresentou saída do tubo de silicone, um dia após a cirurgia. Após uma semana, os outros nove, relataram desaparecimento da epífora. Durante o primeiro mês de seguimento, mais um paciente apresentou perda do tubo de silicone e outro voltou apresentar epífora, por obstrução da fístula lacrimonasal. CONCLUSÕES: A dacriocistorrinostomia assistida por laser diodo, realizada pela via transcanalicular, é novo método para tratamento da obstrução das vias lacrimais. Com o desenvolvimento desta técnica espera-se aumento no índice de sucesso cirúrgico, tanto imediato como a longo prazo.PURPOSE: To describe the technique and initial results of laser-assisted dacryocystorhinostomy performed through the canaliculi. METHODS: Ten patients with nasolacrimal duct obstruction underwent transcanalicular laser-assisted dacryocystorhinostomy. A silicone tube was inserted through the canaliculi and the ostium into the nasal cavity where it will be kept for 6 months. RESULTS: All ten operations were performed without negative occurrences. One patient presented displacement of the silicone tube one day after surgery. Nine of the ten patients reported disappearance of epiphora at the end of the first week following surgery. During the first month, one of these patients presented with epiphora due to obstruction of the lacrimal-nasal fistula and another lost the silicone tube in the first month following surgery. CONCLUSIONS: Transcanalicular laser-assisted dacryocystorhinostomy is a

  11. Low-temperature laser assisted CBE-growth of AlGaAs

    Science.gov (United States)

    Jothilingam, R.; Farrell, T.; Joyce, T. B.; Goodhew, P. J.

    1998-06-01

    We report preliminary studies of low-temperature (335-400°C) chemical beam epitaxial (CBE) growth of Al xGa 1- xAs on GaAs(0 0 1) using triethylgallium (TEG), trimethylaminealane (TMAA) and thermally precracked Arsine (AsH 3) as precursors. We also report results of Ar + laser assisted chemical beam epitaxial growth over the same temperature range. The growth rate for both assisted and unassisted growth as a function of substrate temperature, laser power and precursor beam pressures was determined using laser reflectometry in which the Ar + laser was also used as the probe. In the nonlaser assisted growth Al incorporation is observed to be significantly higher than would be expected at the normal growth temperature of 500°C. With laser assistance the Al concentration, while higher than that at normal growth temperatures, is less than that without laser assistance and the growth rate is higher. These observations, which extended Abernathy's early results to higher nominal Al concentration, are discussed in terms of the relative enhancement of the decomposition of TEG and the alane during laser assistance. Using literature values of the refractive index of AlGaAs alloys at the growth temperature, laser reflectometry was used to monitor both composition and growth rate over a range of growth temperatures. Reflectometry data were compared with the results of Auger Electron Microscopy (AES) and Dektak stylus profiling.

  12. The effects of laser assisted hatching on pregnancy rates

    Directory of Open Access Journals (Sweden)

    Alireza Ghannadi

    2011-01-01

    Full Text Available Background: For infertile women aged over 35 years, failure of the ZP (zona pellucida to rupture is believed to be associated with a decreased implantation rate in in vitro fertilization (IVF or intra cytoplasmic sperm injection (ICSI.Objective: In this research, laser assisted hatching (LAH was offered to patients with advanced maternal age to evaluate a possible benefit.Materials and Methods: Nine hundred thirty two cycles of IVF/ICSI in females were analyzed. Women included in this study were allocated in 4 groups. In group I and II, embryos were cultured and transferred with and without LAH in women aged ≤35, whereas embryos of group III and IV were examined with and without LAH in women aged ≥ 35. Laser manipulations were performed using a suturn-Tm3 system using 2-3 pulses of 0.8 millisecond with 400 voltage duration. The size of the hole made in the zona was measured to be 5-10 μm, depending on the zona thickness of each individual embryo.Results: The performance of LAH significantly increased clinical pregnancy rates in all patients. In group I and II, the chemical (50.99% and 31.61% respectively, clinical (50% and 30.69% respectively and multiple pregnancies (22.27% and 5.94% respectively significantly differ between these groups. In the patients with advanced female age ≥35 the performance of LAH significantly increased chemical (30.12% and clinical pregnancy (27.71% rates compared to whom without LAH (18.96% and 16.37% respectively.Conclusion: Our data demonstrate in the patients who were less than 35 years old, multiple pregnancy rates were significantly increased compared to other groups who aged over 35 years old. In addition benefit of LAH in improving pregnancy rates after IVF or ICSI in women of advanced age (≥35 was shown

  13. The effects of laser assisted hatching on pregnancy rates

    Science.gov (United States)

    Ghannadi, Alireza; Kazerooni, Marjaneh; Jamalzadeh, Fatemeh; Amiri, Sahar; Rostami, Parifar; Absalan, Forouzan

    2011-01-01

    Background: For infertile women aged over 35 years, failure of the ZP (zona pellucida) to rupture is believed to be associated with a decreased implantation rate in in vitro fertilization (IVF) or intra cytoplasmic sperm injection (ICSI). Objective: In this research, laser assisted hatching (LAH) was offered to patients with advanced maternal age to evaluate a possible benefit. Materials and Methods: Nine hundred thirty two cycles of IVF/ICSI in females were analyzed. Women included in this study were allocated in 4 groups. In group I and II, embryos were cultured and transferred with and without LAH in women aged ≤35, whereas embryos of group III and IV were examined with and without LAH in women aged ≥ 35. Laser manipulations were performed using a suturn-Tm3 system using 2-3 pulses of 0.8 millisecond with 400 voltage duration. The size of the hole made in the zona was measured to be 5-10 µm, depending on the zona thickness of each individual embryo. Results: The performance of LAH significantly increased clinical pregnancy rates in all patients. In group I and II, the chemical (50.99% and 31.61% respectively), clinical (50% and 30.69% respectively) and multiple pregnancies (22.27% and 5.94% respectively) significantly differ between these groups. In the patients with advanced female age ≥35 the performance of LAH significantly increased chemical (30.12%) and clinical pregnancy (27.71%) rates compared to whom without LAH (18.96% and 16.37% respectively). Conclusion: Our data demonstrate in the patients who were less than 35 years old, multiple pregnancy rates were significantly increased compared to other groups who aged over 35 years old. In addition benefit of LAH in improving pregnancy rates after IVF or ICSI in women of advanced age (≥35) was shown. PMID:25587254

  14. Nickel-disilicide-assisted excimer laser crystallization of amorphous silicon

    Institute of Scientific and Technical Information of China (English)

    Liao Yan-Ping; Shao Xi-Bin; Gao Feng-Li; Luo Wen-Sheng; Wu Yuan; Fu Guo-Zhu; Jing Hai; Ma Kai

    2006-01-01

    Polycrystalline silicon (poly-Si) thin film has been prepared by means of nickel-disilicide (NiSi2) assisted excimer laser crystallization (ELC). The process to prepare a sample includes two steps. One step consists of the formation of NiSi2 precipitates by heat-treating the dehydrogenated amorphous silicon (a-Si) coated with a thin layer of Ni. And the other step consists of the formation of poly-Si grains by means of ELC. According to the test results of scanning electron microscopy (SEM), another grain growth model named two-interface grain growth has been proposed to contrast with the conventional Ni-metal-induced lateral crystallization (Ni-MILC) model and the ELC model. That is, an additional grain growth interface other than that in conventional ELC is formed, which consists of NiSi2 precipitates and a-Si.The processes for grain growth according to various excimer laser energy densities delivered to the a-Si film have been discussed. It is discovered that grains with needle shape and most of a uniform orientation are formed which grow up with NiSi2 precipitates as seeds. The reason for the formation of such grains which are different from that of Ni-MILCwithout migration of Ni atoms is not clear. Our model and analysis point out a method to prepare grains with needle shape and mostly of a uniform orientation. If such grains are utilized to make thin-film transistor, its characteristics may be improved.

  15. First Demonstration of Laser-Assisted Charge Exchange for Microsecond Duration H- Beams

    Science.gov (United States)

    Cousineau, Sarah; Rakhman, Abdurahim; Kay, Martin; Aleksandrov, Alexander; Danilov, Viatcheslav; Gorlov, Timofey; Liu, Yun; Plum, Michael; Shishlo, Andrei; Johnson, David

    2017-02-01

    This Letter reports on the first demonstration of laser-assisted H- charge exchange for microsecond duration H- beam pulses. Laser-assisted charge exchange injection is a breakthrough technology that overcomes long-standing limitations associated with the traditional method of producing high intensity, time structured beams of protons in accelerators via the use of carbon foils for charge exchange injection. The central theme of this experiment is the demonstration of novel techniques that reduce the laser power requirement to allow high efficiency stripping of microsecond duration beams with commercial laser technology.

  16. Study on the effect of thermal property of metals in ultrasonic-assisted laser machining

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hu Seung; Kim, Gun Woo; Park, Jong Eun [Dept. of Mechanical Engineering, Korea Advanced Institute of Science and Technology, Daejeon (Korea, Republic of); Cho, Sung Hak; Yang, Min Yang; Park, Jong Kweon [Korea Institute of Machinery and Materials, Deajeon (Korea, Republic of)

    2015-08-15

    The laser machining process has been proposed as an advanced process for the selective fabrication of electrodes without a mask. In this study, we adapt laser machining to metals that have different thermal properties. Based on the results, the metals exhibit a different surface morphology, heat-affected zone (HAZ), and a recast layer around the machined surface according to their thermal conductivity, boiling point, and thermal diffusivity. Then, we apply ultrasonic-assisted laser machining to remove the recast layer. The ultrasonic-assisted laser machining exhibits a better surface quality in metals with higher diffusivity than those having lower diffusivity.

  17. Femtosecond laser-assisted lamellar keratoplasty Transplante lamelar auxiliado pelo laser de fentosegundo

    Directory of Open Access Journals (Sweden)

    Hunson Kaz Soong

    2008-08-01

    Full Text Available Lamellar keratoplasty consists of transplanting partial-thickness donor cornea onto a complementary recipient bed. Manual lamellar dissection is technically very difficult, time-consuming, and imprecise. Also, the manually-dissected lamellar interface often has topographical irregularities that may optically degrade the best-corrected visual acuity. The femtosecond clinical laser (IntraLase FS LaserTM, Irvine, CA is a recent innovation that can be programmed to produce bladeless, precise lamellar cuts at any depth with accompanying trephination cuts for both anterior and posterior lamellar transplantion. Posterior laser cuts may be used to assist in deep lamellar endothelial keratoplasty or Descemet's stripping automated endothelial keratoplasty.A ceratoplastia lamelar consiste em transplante de espessura parcial da córnea doadora em um leito receptor complementar. A dissecção lamelar manual é técnica de difícil realização, imprecisa e que demanda tempo. Além disso, a interface lamelar freqüentemente apresenta irregularidade topográfica que pode comprometer a acuidade visual final. O laser clínico "femtosecond" (IntraLase FS LaserTM, Irvine, CA é uma recente inovação que pode ser utilizado para produzir cortes lamelares precisos em qualquer profundidade da córnea, acompanhados de cortes verticais tanto para transplantes lamelares anteriores como posteriores sem a utilização de lâminas. Os cortes posteriores podem ser utilizados para a realização de ceratoplastia endotelial lamelar profunda ou ceratoplastia endotelial com remoção da membrana de Descemet.

  18. Computer-Assisted Experiments with a Laser Diode

    Science.gov (United States)

    Kraftmakher, Yaakov

    2011-01-01

    A laser diode from an inexpensive laser pen (laser pointer) is used in simple experiments. The radiant output power and efficiency of the laser are measured, and polarization of the light beam is shown. The "h/e" ratio is available from the threshold of spontaneous emission. The lasing threshold is found using several methods. With a…

  19. Laser-Assisted in situ Keratomileusis for Hyperopic Defects

    Directory of Open Access Journals (Sweden)

    Ester Novoa Sánchez

    2014-06-01

    Full Text Available Background: currently, LASIK is one of the most common surgical procedures for refractive disorders, including hyperopic defects. Objective: to assess the results of laser-assisted in situ keratomileusis for the treatment of hyperopic defects. Methods: an ambispective study was conducted in 31 eyes (16 patients operated on for hyperopia and simple and compound hyperopic astigmatism in the Refractive Surgery Unit of the Ophthalmology Department of the Hermanos Ameijeiras Hospital between January 2009 and December 2010. Visual acuity with and without optical correction, spherical and astigmatic component before and after surgery and intra- and postoperative complications were analyzed. Results: patients aged 40 to 49 years predominated; compound hyperopic astigmatism was the most frequent defect; the mean uncorrected visual acuity improved and remained stable up to six months after surgery; the average values were 0.87 for moderate defects and 0.79 for high defects with optical correction. The mean sphero-cylindrical component decreased. The efficacy and safety rates were acceptable for moderate and high degrees of the defects and predictability was 100 % for all degrees of ametropia. Conclusion: LASIK is a safe, effective and predictable procedure to treat patients with moderate and high degrees hyperopia and hyperopic astigmatism.

  20. Atomic Force Microscopy Thermally-Assisted Microsampling with Atmospheric Pressure Temperature Ramped Thermal Desorption/Ionization-Mass Spectrometry Analysis.

    Science.gov (United States)

    Hoffmann, William D; Kertesz, Vilmos; Srijanto, Bernadeta R; Van Berkel, Gary J

    2017-02-20

    The use of atomic force microscopy controlled nanothermal analysis probes for reproducible spatially resolved thermally assisted sampling of micrometer-sized areas (ca. 11 × 17 μm wide × 2.4 μm deep) from relatively low number-average molecular weight (Mn mass spectrometric analysis. The procedure and mechanism for material pickup, the sampling reproducibility and sampling size are discussed, and the oligomer distribution information available from slow temperature ramps versus ballistic temperature jumps is presented. For the Mn = 970 P2VP, the Mn and polydispersity index determined from the mass spectrometric data were in line with both the label values from the sample supplier and the value calculated from the simple infusion of a solution of polymer into the commercial atmospheric pressure chemical ionization source on this mass spectrometer. With a P2VP sample of higher Mn (Mn = 2070 and 2970), intact oligomers were still observed (as high as m/z 2793 corresponding to the 26-mer), but a significant abundance of thermolysis products were also observed. In addition, the capability for confident identification of the individual oligomers by slowly ramping the probe temperature and collecting data-dependent tandem mass spectra was also demonstrated. The material type limits to the current sampling and analysis approach as well as possible improvements in nanothermal analysis probe design to enable smaller area sampling and to enable controlled temperature ramps beyond the present upper limit of about 415 °C are also discussed.

  1. Evaluation of laser desorption mass spectrometry and UV accelerated aging of dyes on paper as tools for the evaluation of a questioned document.

    Science.gov (United States)

    Grim, Donzna M; Siegel, Jay; Allison, John

    2002-11-01

    Laser desorption mass spectrometry (LDMS) may be used for the detection and identification of dyes found in inks. Naturally-aged and artificially-aged blue and black ballpoint pen inks containing the cationic dye methyl violet were analyzed on paper. The average molecular weight of the dye sample was calculated from LD mass spectral data and plotted versus time. The resulting aging curves demonstrate that, as dye degradation increases, the average molecular weight of the dye decreases. Typical variables involved in ink aging, such as the type of paper and ink formulation, were investigated. Results show that these variables influence the rate of dye degradation. Furthermore, UV accelerated aging has been developed and tested as an alternative to thermal approaches.

  2. Desorption/Ionization Fluence Thresholds and Improved Mass Spectral Consistency Measured Using a Flattop Laser Profile in the Bioaerosol Mass Spectrometry of Single Bacillus Endospores

    Energy Technology Data Exchange (ETDEWEB)

    Steele, P T; Srivastava, A; Pitesky, M E; Fergenson, D P; Tobias, H J; Gard, E E; Frank, M

    2004-11-30

    Bioaerosol mass spectrometry (BAMS) is being developed to analyze and identify biological aerosols in real-time. Mass spectra of individual Bacillus endospores were measured here with a bipolar aerosol time-of-flight mass spectrometer in which molecular desorption and ionization were produced using a single laser pulse from a Q-switched, frequency-quadrupled Nd:YAG laser that was modified to have an approximately flattop profile. The flattened laser profile allowed the minimum fluence required to desorb and ionize significant numbers of ions from single aerosol particles to be determined. For Bacillus spores this threshold had a mean value of approximately 1 nJ/{micro}m{sup 2} (0.1 J/cm{sup 2}). Thresholds for individual spores, however, could apparently deviate by 20% or more from the mean. Threshold distributions for clumps of MS2 bacteriophage and bovine serum albumin were subsequently determined. Finally, the flattened profile was observed to increase the reproducibility of single spore mass spectra. This is consistent with the general conclusions of our earlier paper on the fluence dependence of single spore mass spectra and is particularly significant because it is expected to enable more robust differentiation and identification of single bioaerosol particles.

  3. Preliminary results of femtosecond laser-assisted descemet stripping endothelial keratoplasty.

    NARCIS (Netherlands)

    Cheng, Y.Y.; Hendrikse, F.; Pels, E.; Wijdh, R.J.; Cleynenbreugel, H. van; Eggink, C.A.; Rij, G. van; Rijneveld, W.J.; Nuijts, R.M.

    2008-01-01

    OBJECTIVE: To evaluate the preliminary visual results of femtosecond laser-assisted Descemet stripping endothelial keratoplasty (FS-DSEK). METHODS: We prospectively analyzed results of 20 consecutive patients with Fuchs endothelial dystrophy or aphakic/pseudophakic bullous keratopathy who underwent

  4. Fractionation of a coal tar pitch by ultra-filtration, and characterization by size exclusion chromatography, UV-fluorescence and laser desorption-mass spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Anthe George; Trevor J. Morgan; Patricia Alvarez; Marcos Millan; Alan A. Herod; Rafael Kandiyoti [Imperial College London, London (United Kingdom). Department of Chemical Engineering

    2010-10-15

    Ultra-filtration (UF) provides a new way of generating narrow bands of sample in the fractionation of coal and petroleum-derived liquids. It allows larger quantities of high-mass fractions to be recovered, making more detailed investigations possible, through the use of techniques requiring larger amounts of sample. In this work, UF-separated fractions have been used to study molecular mass distributions of a coal tar pitch, used as laboratory standard. The pitch was fractionated by solvent solubility into three fractions. These were further fractionated by ultra-filtration, using membranes specific to protein molecular sizes classed as '1 kDa', '5 kDa', '10 kDa' and '100 kDa'. Planar chromatography was used as a tertiary fractionation method, to sub-divide the UF-fractions. The various fractions were examined by size exclusion chromatography (SEC), UV-fluorescence spectroscopy and laser desorption-mass spectrometry (LD-MS). There was good agreement between mass estimates based on SEC and LD-MS of the smaller UF-fractions, with evidence for the presence of material with molecular masses ranging between 800 and 10,000 u. Examining the largest UF fractions of the pitch-pyridine-insoluble sample also gave clear evidence for material with molecular masses above 10,000 u. Taken together, however, the LD-MS data showed progressively diminishing differences, as the sizes of the UF membranes, and the likely molecular masses of the sample fractions, increased. One likely explanation is incomplete sampling during the laser desorption procedure. The evidence suggests that the upper mass limit detectable for these and similar samples by LD-MS has been reached. Despite these reservations, LD-MS appears as the best method to date, for investigating the mass ranges of samples derived from coal tar pitch and heavy petroleum fractions. 51 refs., 6 figs., 3 tabs.

  5. Translational medicine in the field of ablative fractional laser (AFXL)-assisted drug delivery

    DEFF Research Database (Denmark)

    Haedersdal, Merete; Erlendsson, Andrés M; Paasch, Uwe

    2016-01-01

    BACKGROUND: Ablative fractional lasers enhance uptake of topical therapeutics and the concept of fractional laser-assisted drug delivery has now been taken into clinical practice. OBJECTIVES: We systematically reviewed preclinical data and clinical evidence for fractional lasers to enhance drug....... The highest level of evidence was reached for actinic keratoses treated with methylaminolevulinate for photodynamic therapy (level IB, 5 randomized controlled trials), substantiating superior and long-lasting efficacy versus conventional photodynamic therapy. No adverse events were reported, but ablative...... fractional laser-assisted drug delivery implies risks of systemic drug absorption, especially when performed over large skin areas. CONCLUSIONS: Fractional laser-assisted drug delivery is beneficial in enhancing preclinical and clinical outcomes for certain skin conditions....

  6. Realizing the Harper Hamiltonian with laser-assisted tunneling in optical lattices.

    Science.gov (United States)

    Miyake, Hirokazu; Siviloglou, Georgios A; Kennedy, Colin J; Burton, William Cody; Ketterle, Wolfgang

    2013-11-01

    We experimentally implement the Harper Hamiltonian for neutral particles in optical lattices using laser-assisted tunneling and a potential energy gradient provided by gravity or magnetic field gradients. This Hamiltonian describes the motion of charged particles in strong magnetic fields. Laser-assisted tunneling processes are characterized by studying the expansion of the atoms in the lattice. The band structure of this Hamilt