Sample records for samples preparation methods

  1. Fluidics platform and method for sample preparation (United States)

    Benner, Henry W.; Dzenitis, John M.


    Provided herein are fluidics platforms and related methods for performing integrated sample collection and solid-phase extraction of a target component of the sample all in one tube. The fluidics platform comprises a pump, particles for solid-phase extraction and a particle-holding means. The method comprises contacting the sample with one or more reagents in a pump, coupling a particle-holding means to the pump and expelling the waste out of the pump while the particle-holding means retains the particles inside the pump. The fluidics platform and methods herein described allow solid-phase extraction without pipetting and centrifugation.

  2. Global metabolite analysis of yeast: evaluation of sample preparation methods

    DEFF Research Database (Denmark)

    Villas-Bôas, Silas Granato; Højer-Pedersen, Jesper; Åkesson, Mats Fredrik


    Sample preparation is considered one of the limiting steps in microbial metabolome analysis. Eukaryotes and prokaryotes behave very differently during the several steps of classical sample preparation methods for analysis of metabolites. Even within the eukaryote kingdom there is a vast diversity...... of cell structures that make it imprudent to blindly adopt protocols that were designed for a specific group of microorganisms. We have therefore reviewed and evaluated the whole sample preparation procedures for analysis of yeast metabolites. Our focus has been on the current needs in metabolome analysis......, which is the analysis of a large number of metabolites with very diverse chemical and physical properties. This work reports the leakage of intracellular metabolites observed during quenching yeast cells with cold methanol solution, the efficacy of six different methods for the extraction...

  3. Effect of method of sample preparation on ruminal in situ ...

    African Journals Online (AJOL)

    The objective of this study was to investigate the effect of method of sample preparation on the degradation kinetics of herbage when applying the in situ technique. Ryegrass (Lolium multiflorum cv. Midmar) was harvested at three and four weeks after cutting and fertilizing with 200 kg nitrogen (N)/ha. Freshly cut herbage ...

  4. Fluidics platform and method for sample preparation and analysis (United States)

    Benner, W. Henry; Dzenitis, John M.; Bennet, William J.; Baker, Brian R.


    Herein provided are fluidics platform and method for sample preparation and analysis. The fluidics platform is capable of analyzing DNA from blood samples using amplification assays such as polymerase-chain-reaction assays and loop-mediated-isothermal-amplification assays. The fluidics platform can also be used for other types of assays and analyzes. In some embodiments, a sample in a sealed tube can be inserted directly. The following isolation, detection, and analyzes can be performed without a user's intervention. The disclosed platform may also comprises a sample preparation system with a magnetic actuator, a heater, and an air-drying mechanism, and fluid manipulation processes for extraction, washing, elution, assay assembly, assay detection, and cleaning after reactions and between samples.

  5. Modern methods of sample preparation for GC analysis

    NARCIS (Netherlands)

    de Koning, S.; Janssen, H.-G.; Brinkman, U.A.Th.


    Today, a wide variety of techniques is available for the preparation of (semi-) solid, liquid and gaseous samples, prior to their instrumental analysis by means of capillary gas chromatography (GC) or, increasingly, comprehensive two-dimensional GC (GC × GC). In the past two decades, a large number

  6. A Method for Microalgae Proteomics Analysis Based on Modified Filter-Aided Sample Preparation. (United States)

    Li, Song; Cao, Xupeng; Wang, Yan; Zhu, Zhen; Zhang, Haowei; Xue, Song; Tian, Jing


    With the fast development of microalgal biofuel researches, the proteomics studies of microalgae increased quickly. A filter-aided sample preparation (FASP) method is widely used proteomics sample preparation method since 2009. Here, a method of microalgae proteomics analysis based on modified filter-aided sample preparation (mFASP) was described to meet the characteristics of microalgae cells and eliminate the error caused by over-alkylation. Using Chlamydomonas reinhardtii as the model, the prepared sample was tested by standard LC-MS/MS and compared with the previous reports. The results showed mFASP is suitable for most of occasions of microalgae proteomics studies.

  7. Final Report for X-ray Diffraction Sample Preparation Method Development

    Energy Technology Data Exchange (ETDEWEB)

    Ely, T. M.; Meznarich, H. K.; Valero, T.


    WRPS-1500790, “X-ray Diffraction Saltcake Sample Preparation Method Development Plan/Procedure,” was originally prepared with the intent of improving the specimen preparation methodology used to generate saltcake specimens suitable for XRD-based solid phase characterization. At the time that this test plan document was originally developed, packed powder in cavity supports with collodion binder was the established XRD specimen preparation method. An alternate specimen preparation method less vulnerable, if not completely invulnerable to preferred orientation effects, was desired as a replacement for the method.

  8. Sample preparation method for glass welding by ultrashort laser pulses yields higher seam strength. (United States)

    Cvecek, K; Miyamoto, I; Strauss, J; Wolf, M; Frick, T; Schmidt, M


    Glass welding by ultrashort laser pulses allows joining without the need of an absorber or a preheating and postheating process. However, cracks generated during the welding process substantially impair the joining strength of the welding seams. In this paper a sample preparation method is described that prevents the formation of cracks. The measured joining strength of samples prepared by this method is substantially higher than previously reported values.

  9. Electrodeposition as an alternate method for preparation of environmental samples for iodide by AMS

    Energy Technology Data Exchange (ETDEWEB)

    Adamic, M.L., E-mail: [Idaho National Laboratory, P.O. Box 1625, Idaho Falls, ID 83402 (United States); Lister, T.E.; Dufek, E.J.; Jenson, D.D.; Olson, J.E. [Idaho National Laboratory, P.O. Box 1625, Idaho Falls, ID 83402 (United States); Vockenhuber, C. [Laboratory of Ion Beam Physics, ETH Zurich, Otto-Stern-Weg 5, 8093 Zurich (Switzerland); Watrous, M.G. [Idaho National Laboratory, P.O. Box 1625, Idaho Falls, ID 83402 (United States)


    This paper presents an evaluation of an alternate method for preparing environmental samples for {sup 129}I analysis by accelerator mass spectrometry (AMS) at Idaho National Laboratory. The optimal sample preparation method is characterized by ease of preparation, capability of processing very small quantities of iodide, and ease of loading into a cathode. Electrodeposition of iodide on a silver wire was evaluated using these criteria. This study indicates that the electrochemically-formed silver iodide deposits produce ion currents similar to those from precipitated silver iodide for the same sample mass. Precipitated silver iodide samples are usually mixed with niobium or silver powder prior to loading in a cathode. Using electrodeposition, the silver is already mixed with the sample and can simply be picked up with tweezers, placed in the sample die, and pressed into a cathode. The major advantage of this method is that the silver wire/electrodeposited silver iodide is much easier to load into a cathode.

  10. Preparation method matters: Aiming at higher NPP diversity and representativeness in sediment samples

    DEFF Research Database (Denmark)

    Enevold, Renée; Odgaard, Bent Vad


    of palynology in archaeological and forensic sciences. NPPs in anthropogenic soils and archaeological samples may be numerous in types as well as in abundance. However, preparing these soil samples with methods based on acid digestion potentially biases NPP assemblages because of differential damage or even...... dissolution of microfossils. In spite of this potential bias standard preparation procedures for pollen analysis have, in most cases without modification, generally been applied to palynological samples used for NPP analysis. We review briefly the advantages of high diversity NPP-analysis and preparation...

  11. Spin column extraction as a new sample preparation method in bioanalysis. (United States)

    Namera, Akira; Saito, Takashi


    Sample preparation is important in obtaining accurate data for qualification and quantification in bioanalysis. We have recently focused on monolithic silica for high-throughput analysis. These extraction processes - using monolithic silica packed in spin column - such as sample loading, washing and elution, are executed by centrifugation. There are several possibilities such as on-column derivatization for the determination of amines or carboxylic acids in the sample. The spin column extraction reduces the sample preparation time required for determination of drugs and other chemicals in biological materials and increases productivity in bioanalysis. We expect spin column extraction to become the mainstream method of sample processing in the future.

  12. A proteomics sample preparation method for mature, recalcitrant leaves of perennial plants. (United States)

    Gang, Deng; Xinyue, Zhong; Na, Zhang; Chengying, Lao; Bo, Wang; Dingxiang, Peng; Lijun, Liu


    Sample preparation is key to the success of proteomics studies. In the present study, two sample preparation methods were tested for their suitability on the mature, recalcitrant leaves of six representative perennial plants (grape, plum, pear, peach, orange, and ramie). An improved sample preparation method was obtained: Tris and Triton X-100 were added together instead of CHAPS to the lysis buffer, and a 20% TCA-water solution and 100% precooled acetone were added after the protein extraction for the further purification of protein. This method effectively eliminates nonprotein impurities and obtains a clear two-dimensional gel electrophoresis array. The method facilitates the separation of high-molecular-weight proteins and increases the resolution of low-abundance proteins. This method provides a widely applicable and economically feasible technology for the proteomic study of the mature, recalcitrant leaves of perennial plants.

  13. A proteomics sample preparation method for mature, recalcitrant leaves of perennial plants.

    Directory of Open Access Journals (Sweden)

    Deng Gang

    Full Text Available Sample preparation is key to the success of proteomics studies. In the present study, two sample preparation methods were tested for their suitability on the mature, recalcitrant leaves of six representative perennial plants (grape, plum, pear, peach, orange, and ramie. An improved sample preparation method was obtained: Tris and Triton X-100 were added together instead of CHAPS to the lysis buffer, and a 20% TCA-water solution and 100% precooled acetone were added after the protein extraction for the further purification of protein. This method effectively eliminates nonprotein impurities and obtains a clear two-dimensional gel electrophoresis array. The method facilitates the separation of high-molecular-weight proteins and increases the resolution of low-abundance proteins. This method provides a widely applicable and economically feasible technology for the proteomic study of the mature, recalcitrant leaves of perennial plants.

  14. Analysis of aroma compounds of Roselle by Dynamic Headspace Sampling using different preparation methods

    DEFF Research Database (Denmark)

    Juhari, Nurul Hanisah Binti; Varming, Camilla; Petersen, Mikael Agerlin


    The influence of different methods of sample preparation on the aroma profiles of dried Roselle (Hibiscus sabdariffa) was studied. Least amounts of aroma compounds were recovered by analysis of whole dry calyxes (WD) followed by ground dry (GD), blended together with water (BTW), and ground...... and then mixed with water (GMW). The highest number of aroma compounds was found in Roselle treated in water bath (2hr/40°C) (GMWKB). GMW was chosen as the preparation method because it was shown to be an efficient extraction method without the possibility of excessive chemical changes of the sample....

  15. Sample preparation method for the combined extraction of ethyl glucuronide and drugs of abuse in hair. (United States)

    Meier, Ulf; Briellmann, Thomas; Scheurer, Eva; Dussy, Franz


    Often in hair analysis, a small hair sample is available while the analysis of a multitude of structurally diverse substances with different concentration ranges is demanded. The analysis of the different substances often requires different sample preparation methods, increasing the amount of required hair sample. When segmental hair analysis is necessary, the amount of hair sample needed is further increased. Therefore, the required sample amount for a full analysis can quickly exceed what is available. To combat this problem, a method for the combined hair sample preparation using a single extraction procedure for analysis of ethyl glucuronide with liquid chromatography-multistage fragmentation mass spectrometry/multiple reaction monitoring (LC-MS3 /MRM) and common drugs of abuse with LC-MRM was developed. The combined sample preparation is achieved by separating ethyl glucuronide from the drugs of abuse into separate extracts by fractionation in the solid-phase extraction step during sample clean-up. A full validation for all substances for the parameters selectivity, linearity, limit of detection, limit of quantification, accuracy, precision, matrix effects, and recovery was successfully completed. The following drugs of abuse were included in the method: Amphetamine; methamphetamine; 3,4-methylenedioxy-N-methylamphetamine (MDMA); 3,4-methylenedioxyamphetamine (MDA); 3,4-methylenedioxy-N-ethylamphetamine (MDE); morphine; 6-monoacetylmorphine; codeine; acetylcodeine; cocaine; benzoylecgonine; norcocaine; cocaethylene; methadone; 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) and methylphenidate. In conclusion, as only 1 sample preparation is needed with 1 aliquot of hair, the presented sample preparation allows an optimal analysis of both ethyl glucuronide and of the drugs of abuse, even when the sample amount is a limiting factor. Copyright © 2017 John Wiley & Sons, Ltd.

  16. Effect of sample preparation methods on photometric determination of the tellurium and cobalt content in the samples of copper concentrates

    Directory of Open Access Journals (Sweden)

    Viktoriya Butenko


    Full Text Available Methods of determination of cobalt and nickel in copper concentrates currently used in factory laboratories are very labor intensive and time consuming. The limiting stage of the analysis is preliminary chemical sample preparation. Carrying out the decomposition process of industrial samples with concentrated mineral acids in open systems does not allow to improve the metrological characteristics of the methods, for this reason improvement the methods of sample preparation is quite relevant and has a practical interest. The work was dedicated to the determination of the optimal conditions of preliminary chemical preparation of copper concentrate samples for the subsequent determination of cobalt and tellurium in the obtained solution using tellurium-spectrophotometric method. Decomposition of the samples was carried out by acid dissolving in individual mineral acids and their mixtures by heating in an open system as well as by using ultrasonification and microwave radiation in a closed system. In order to select the optimal conditions for the decomposition of the samples in a closed system the phase contact time and ultrasonic generator’s power were varied. Intensification of the processes of decomposition of copper concentrates with nitric acid (1:1, ultrasound and microwave radiation allowed to transfer quantitatively cobalt and tellurium into solution spending 20 and 30 min respectively. This reduced the amount of reactants used and improved the accuracy of determination by running the process in strictly identical conditions.

  17. Error baseline rates of five sample preparation methods used to characterize RNA virus populations. (United States)

    Kugelman, Jeffrey R; Wiley, Michael R; Nagle, Elyse R; Reyes, Daniel; Pfeffer, Brad P; Kuhn, Jens H; Sanchez-Lockhart, Mariano; Palacios, Gustavo F


    Individual RNA viruses typically occur as populations of genomes that differ slightly from each other due to mutations introduced by the error-prone viral polymerase. Understanding the variability of RNA virus genome populations is critical for understanding virus evolution because individual mutant genomes may gain evolutionary selective advantages and give rise to dominant subpopulations, possibly even leading to the emergence of viruses resistant to medical countermeasures. Reverse transcription of virus genome populations followed by next-generation sequencing is the only available method to characterize variation for RNA viruses. However, both steps may lead to the introduction of artificial mutations, thereby skewing the data. To better understand how such errors are introduced during sample preparation, we determined and compared error baseline rates of five different sample preparation methods by analyzing in vitro transcribed Ebola virus RNA from an artificial plasmid-based system. These methods included: shotgun sequencing from plasmid DNA or in vitro transcribed RNA as a basic "no amplification" method, amplicon sequencing from the plasmid DNA or in vitro transcribed RNA as a "targeted" amplification method, sequence-independent single-primer amplification (SISPA) as a "random" amplification method, rolling circle reverse transcription sequencing (CirSeq) as an advanced "no amplification" method, and Illumina TruSeq RNA Access as a "targeted" enrichment method. The measured error frequencies indicate that RNA Access offers the best tradeoff between sensitivity and sample preparation error (1.4-5) of all compared methods.

  18. [Preparation of sub-standard samples and XRF analytical method of powder non-metallic minerals]. (United States)

    Kong, Qin; Chen, Lei; Wang, Ling


    In order to solve the problem that standard samples of non-metallic minerals are not satisfactory in practical work by X-ray fluorescence spectrometer (XRF) analysis with pressed powder pellet, a method was studied how to make sub-standard samples according to standard samples of non-metallic minerals and to determine how they can adapt to analysis of mineral powder samples, taking the K-feldspar ore in Ebian-Wudu, Sichuan as an example. Based on the characteristic analysis of K-feldspar ore and the standard samples by X-ray diffraction (XRD) and chemical methods, combined with the principle of the same or similar between the sub-standard samples and unknown samples, the experiment developed the method of preparation of sub-standard samples: both of the two samples above mentioned should have the same kind of minerals and the similar chemical components, adapt mineral processing, and benefit making working curve. Under the optimum experimental conditions, a method for determination of SiO2, Al2O3, Fe2O3, TiO2, CaO, MgO, K2O and Na2O of K-feldspar ore by XRF was established. Thedetermination results are in good agreement with classical chemical methods, which indicates that this method was accurate.


    DEFF Research Database (Denmark)


    The invention relates to a method for preparing a substrate (105a) comprising a sample reception area (110) and a sensing area (111). The method comprises the steps of: 1) applying a sample on the sample reception area; 2) rotating the substrate around a predetermined axis; 3) during rotation......, at least part of the liquid travels from the sample reception area to the sensing area due to capillary forces acting between the liquid and the substrate; and 4) removing the wave of particles and liquid formed at one end of the substrate. The sensing area is closer to the predetermined axis than...... the sample reception area. The sample comprises a liquid part and particles suspended therein....

  20. Cytotoxicity of Light-Cured Dental Materials according to Different Sample Preparation Methods

    Directory of Open Access Journals (Sweden)

    Myung-Jin Lee


    Full Text Available Dental light-cured resins can undergo different degrees of polymerization when applied in vivo. When polymerization is incomplete, toxic monomers may be released into the oral cavity. The present study assessed the cytotoxicity of different materials, using sample preparation methods that mirror clinical conditions. Composite and bonding resins were used and divided into four groups according to sample preparation method: uncured; directly cured samples, which were cured after being placed on solidified agar; post-cured samples were polymerized before being placed on agar; and “removed unreacted layer” samples had their oxygen-inhibition layer removed after polymerization. Cytotoxicity was evaluated using an agar diffusion test, MTT assay, and confocal microscopy. Uncured samples were the most cytotoxic, while removed unreacted layer samples were the least cytotoxic (p < 0.05. In the MTT assay, cell viability increased significantly in every group as the concentration of the extracts decreased (p < 0.05. Extracts from post-cured and removed unreacted layer samples of bonding resin were less toxic than post-cured and removed unreacted layer samples of composite resin. Removal of the oxygen-inhibition layer resulted in the lowest cytotoxicity. Clinicians should remove unreacted monomers on the resin surface immediately after restoring teeth with light-curing resin to improve the restoration biocompatibility.

  1. A simple sample preparation method for measuring amoxicillin in human plasma by hollow fiber centrifugal ultrafiltration. (United States)

    Dong, Wei-Chong; Hou, Zi-Li; Jiang, Xin-Hui; Jiang, Ye


    A simple sample preparation method has been developed for the determination of amoxicillin in human plasma by hollow fiber centrifugal ultrafiltration (HF-CF-UF). A 400-μL plasma sample was placed directly into the HF-CF-UF device, which consisited of a slim glass tube and a U-shaped hollow fiber. After centrifugation at 1.25 × 10(3) g for 10 min, the filtrate was withdrawn from the hollow fiber and 20 µL was directly injected into the high-performance liquid chromatography (HPLC) for analysis. The calibration curve was linear over the range of 0.1-20 µg/mL (r = 0.9996) and the limit of detection was as low as 0.025 µg/mL. The average recovery and absolute recovery were 99.9% and 84.5%, respectively. Both the intra-day and inter-day precisions (relative standard deviation) were less than 3.1% for three concentrations (0.25, 2.5 and 10 µg/mL). The sample preparation process was simplified. Only after a single centrifugal ultrafiltration can the filtrate be injected directly into HPLC. The present method is simple, sensitive and accurate. It could be effective for the analysis of biological samples with high protein contents, especially for the biopharmaceutical analysis of drugs that use traditional isolation techniques for sample preparation such as the protein precipitation method.

  2. A sample preparation method for recovering suppressed analyte ions in MALDI TOF MS. (United States)

    Lou, Xianwen; de Waal, Bas F M; Milroy, Lech-Gustav; van Dongen, Joost L J


    In matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS), analyte signals can be substantially suppressed by other compounds in the sample. In this technical note, we describe a modified thin-layer sample preparation method that significantly reduces the analyte suppression effect (ASE). In our method, analytes are deposited on top of the surface of matrix preloaded on the MALDI plate. To prevent embedding of analyte into the matrix crystals, the sample solution were prepared without matrix and efforts were taken not to re-dissolve the preloaded matrix. The results with model mixtures of peptides, synthetic polymers and lipids show that detection of analyte ions, which were completely suppressed using the conventional dried-droplet method, could be effectively recovered by using our method. Our findings suggest that the incorporation of analytes in the matrix crystals has an important contributory effect on ASE. By reducing ASE, our method should be useful for the direct MALDI MS analysis of multicomponent mixtures. Copyright © 2015 John Wiley & Sons, Ltd.

  3. Filter-Aided Sample Preparation: The Versatile and Efficient Method for Proteomic Analysis. (United States)

    Wiśniewski, J R


    Filter-aided sample preparation (FASP) is a versatile and efficient way of processing protein extracts for bottom-up proteomic analysis. The method repurposes centrifugal ultrafiltration concentrators for removal of detergents, protein cleavage, and isolation of pure peptide fractions. FASP can be used for protein cleavage with different proteinases either with single enzymes or in a mode of successive multienzyme digestion (MED)-FASP. The FASP methods are useful for processing of samples ranging in their sizes from submicrogram to several milligram amounts of total protein. They also allow peptide fractionation, and isolation and quantitation of total RNA and DNA acid contents. This chapter describes principles, limitations, and applications of FASP. Additionally detailed FASP and MED-FASP protocols are provided. © 2017 Elsevier Inc. All rights reserved.

  4. Comparison of sample preparation methods for detection of Chlamydia pneumoniae in bronchoalveolar lavage fluid by PCR. (United States)

    Maass, M; Dalhoff, K


    Amplification inhibitors can lead to false-negative results for PCR. In order to evaluate the reliability of PCR for the detection of Chlamydia pneumoniae, the presence of PCR inhibitors in 75 bronchoalveolar lavage specimens was assessed after treatment by various sample preparation methods. Specimens were collected from patients with acute respiratory infections, including four cases of proven C. pneumoniae infection. Substances inhibitory to the amplification of chlamydial DNA continued to be present in 12% of the samples treated according to the commonly used single-step proteinase K digestion and in 31% of the samples processed by heat treatment. However, the complexing of DNA-contaminating proteins and polysaccharides from digested specimens to cetyltrimethylammonium bromide (CTAB) followed by DNA extraction efficiently removed inhibitors from all experimental samples and provided subsequent identification of all positive clinical samples by PCR. The CTAB method and proteinase K treatment had comparable detection limits of approximately 0.01 inclusion-forming units. CTAB-based DNA purification of respiratory specimens is recommended to increase the diagnostic sensitivity of PCR and confidence in negative results. Images PMID:7814512

  5. Preparation of Biological Samples Containing Metoprolol and Bisoprolol for Applying Methods for Quantitative Analysis

    Directory of Open Access Journals (Sweden)

    Corina Mahu Ştefania


    Full Text Available Arterial hypertension is a complex disease with many serious complications, representing a leading cause of mortality. Selective beta-blockers such as metoprolol and bisoprolol are frequently used in the management of hypertension. Numerous analytical methods have been developed for the determination of these substances in biological fluids, such as liquid chromatography coupled with mass spectrometry, gas chromatography coupled with mass spectrometry, high performance liquid chromatography. Due to the complex composition of biological fluids a biological sample pre-treatment before the use of the method for quantitative determination is required in order to remove proteins and potential interferences. The most commonly used methods for processing biological samples containing metoprolol and bisoprolol were identified through a thorough literature search using PubMed, ScienceDirect, and Willey Journals databases. Articles published between years 2005-2015 were reviewed. Protein precipitation, liquid-liquid extraction and solid phase extraction are the main techniques for the extraction of these drugs from plasma, serum, whole blood and urine samples. In addition, numerous other techniques have been developed for the preparation of biological samples, such as dispersive liquid-liquid microextraction, carrier-mediated liquid phase microextraction, hollow fiber-protected liquid phase microextraction, on-line molecularly imprinted solid phase extraction. The analysis of metoprolol and bisoprolol in human plasma, urine and other biological fluids provides important information in clinical and toxicological trials, thus requiring the application of appropriate extraction techniques for the detection of these antihypertensive substances at nanogram and picogram levels.

  6. Final LDRD report : development of sample preparation methods for ChIPMA-based imaging mass spectrometry of tissue samples.

    Energy Technology Data Exchange (ETDEWEB)

    Maharrey, Sean P.; Highley, Aaron M.; Behrens, Richard, Jr.; Wiese-Smith, Deneille


    The objective of this short-term LDRD project was to acquire the tools needed to use our chemical imaging precision mass analyzer (ChIPMA) instrument to analyze tissue samples. This effort was an outgrowth of discussions with oncologists on the need to find the cellular origin of signals in mass spectra of serum samples, which provide biomarkers for ovarian cancer. The ultimate goal would be to collect chemical images of biopsy samples allowing the chemical images of diseased and nondiseased sections of a sample to be compared. The equipment needed to prepare tissue samples have been acquired and built. This equipment includes an cyro-ultramicrotome for preparing thin sections of samples and a coating unit. The coating unit uses an electrospray system to deposit small droplets of a UV-photo absorbing compound on the surface of the tissue samples. Both units are operational. The tissue sample must be coated with the organic compound to enable matrix assisted laser desorption/ionization (MALDI) and matrix enhanced secondary ion mass spectrometry (ME-SIMS) measurements with the ChIPMA instrument Initial plans to test the sample preparation using human tissue samples required development of administrative procedures beyond the scope of this LDRD. Hence, it was decided to make two types of measurements: (1) Testing the spatial resolution of ME-SIMS by preparing a substrate coated with a mixture of an organic matrix and a bio standard and etching a defined pattern in the coating using a liquid metal ion beam, and (2) preparing and imaging C. elegans worms. Difficulties arose in sectioning the C. elegans for analysis and funds and time to overcome these difficulties were not available in this project. The facilities are now available for preparing biological samples for analysis with the ChIPMA instrument. Some further investment of time and resources in sample preparation should make this a useful tool for chemical imaging applications.

  7. A novel sample preparation method using rapid nonheated saponification method for the determination of cholesterol in emulsified foods. (United States)

    Jeong, In-Seek; Kwak, Byung-Man; Ahn, Jang-Hyuk; Leem, Donggil; Yoon, Taehyung; Yoon, Changyong; Jeong, Jayoung; Park, Jung-Min; Kim, Jin-Man


    In this study, nonheated saponification was employed as a novel, rapid, and easy sample preparation method for the determination of cholesterol in emulsified foods. Cholesterol content was analyzed using gas chromatography with a flame ionization detector (GC-FID). The cholesterol extraction method was optimized for maximum recovery from baby food and infant formula. Under these conditions, the optimum extraction solvent was 10 mL ethyl ether per 1 to 2 g sample, and the saponification solution was 0.2 mL KOH in methanol. The cholesterol content in the products was determined to be within the certified range of certified reference materials (CRMs), NIST SRM 1544 and SRM 1849. The results of the recovery test performed using spiked materials were in the range of 98.24% to 99.45% with an relative standard devitation (RSD) between 0.83% and 1.61%. This method could be used to reduce sample pretreatment time and is expected to provide an accurate determination of cholesterol in emulsified food matrices such as infant formula and baby food. A novel, rapid, and easy sample preparation method using nonheated saponification was developed for cholesterol detection in emulsified foods. Recovery tests of CRMs were satisfactory, and the recoveries of spiked materials were accurate and precise. This method was effective and decreased the time required for analysis by 5-fold compared to the official method. © 2012 Institute of Food Technologists®

  8. Evaluation of sample preparation methods and optimization of nickel determination in vegetable tissues

    Directory of Open Access Journals (Sweden)

    Rodrigo Fernando dos Santos Salazar


    Full Text Available Nickel, although essential to plants, may be toxic to plants and animals. It is mainly assimilated by food ingestion. However, information about the average levels of elements (including Ni in edible vegetables from different regions is still scarce in Brazil. The objectives of this study were to: (a evaluate and optimize a method for preparation of vegetable tissue samples for Ni determination; (b optimize the analytical procedures for determination by Flame Atomic Absorption Spectrometry (FAAS and by Electrothermal Atomic Absorption (ETAAS in vegetable samples and (c determine the Ni concentration in vegetables consumed in the cities of Lorena and Taubaté in the Vale do Paraíba, State of São Paulo, Brazil. By means of the analytical technique for determination by ETAAS or FAAS, the results were validated by the test of analyte addition and recovery. The most viable method tested for quantification of this element was HClO4-HNO3 wet digestion. All samples but carrot tissue collected in Lorena contained Ni levels above the permitted by the Brazilian Ministry of Health. The most disturbing results, requiring more detailed studies, were the Ni concentrations measured in carrot samples from Taubaté, where levels were five times higher than permitted by Brazilian regulations.

  9. An efficient and sensitive method for preparing cDNA libraries from scarce biological samples. (United States)

    Sterling, Catherine H; Veksler-Lublinsky, Isana; Ambros, Victor


    The preparation and high-throughput sequencing of cDNA libraries from samples of small RNA is a powerful tool to quantify known small RNAs (such as microRNAs) and to discover novel RNA species. Interest in identifying the small RNA repertoire present in tissues and in biofluids has grown substantially with the findings that small RNAs can serve as indicators of biological conditions and disease states. Here we describe a novel and straightforward method to clone cDNA libraries from small quantities of input RNA. This method permits the generation of cDNA libraries from sub-picogram quantities of RNA robustly, efficiently and reproducibly. We demonstrate that the method provides a significant improvement in sensitivity compared to previous cloning methods while maintaining reproducible identification of diverse small RNA species. This method should have widespread applications in a variety of contexts, including biomarker discovery from scarce samples of human tissue or body fluids. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  10. Suspension trapping (STrap) sample preparation method for bottom-up proteomics analysis. (United States)

    Zougman, Alexandre; Selby, Peter J; Banks, Rosamonde E


    Despite recent developments in bottom-up proteomics, the need still exists in a fast, uncomplicated, and robust method for comprehensive sample processing especially when applied to low protein amounts. The suspension trapping method combines the advantage of efficient SDS-based protein extraction with rapid detergent removal, reactor-type protein digestion, and peptide cleanup. Proteins are solubilized in SDS. The sample is acidified and introduced into the suspension trapping tip incorporating the depth filter and hydrophobic compartments, filled with the neutral pH methanolic solution. The instantly formed fine protein suspension is trapped in the depth filter stack-this crucial step is aimed at separating the particulate matter in space. SDS and other contaminants are removed in the flow-through, and a protease is introduced. Following the digestion, the peptides are cleaned up using the tip's hydrophobic part. The methodology allows processing of protein loads down to the low microgram/submicrogram levels. The detergent removal takes about 5 min, whereas the tryptic proteolysis of a cellular lysate is complete in as little as 30 min. We have successfully utilized the method for analysis of cellular lysates, enriched membrane preparations, and immunoprecipitates. We expect that due to its robustness and simplicity, the method will become an essential proteomics tool. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Evaluation of sample preparation methods for water activity determination in jerky and kippered beef: a research note. (United States)

    Harper, N M; Getty, K J K; Boyle, E A E


    Commercially available packaged whole muscle beef jerky, chopped and formed beef jerky, and kippered beef steak were obtained from retail stores to determine the effect of two sample preparation methods on water activity (a(w)). Intact samples were prepared by cutting product into a hexagonal shape with a 3.2 cm diameter. Diced samples were prepared by cutting the product into 0.4 x 0.4 cm squares. Whole muscle jerky a(w) was higher (0.016 units; P0.05) in a(w) levels between the two preparation methods for chopped and formed jerky or kippered beef steak. An intact sample preparation method is recommended for a(w) determination of whole muscle jerky to obtain a more conservative value, especially if a(w) is near the margin of safety. Copyright (c) 2010 The American Meat Science Association. Published by Elsevier Ltd. All rights reserved.

  12. Contemporary Sample Preparation Methods for the Detection of Ignitable Liquids in Suspect Arson Cases. (United States)

    Bertsch, W; Ren, Q


    The isolation of ignitable liquid components, usually petroleum-based distillates from fire debris, is an important step in deciding whether a fire is of natural or incendiary origin. Steady progress has been made to develop sample preparation methods capable of enriching target analytes in high yield and within a short period of time. Heated headspace enrichment methods are currently most widely used. There are several variations of this basic technique. Carbon-based adsorbents are most popular. They come in different forms and shapes, including a flat sheet of polymer, impregnated with carbon particles. The analyst cuts a small strip from this sheet and suspends it in the heated headspace above the debris sample. The volatiles adsorb onto the carbon surface, eventually reaching an equilibrium condition. The process is usually carried out in an oven. This convenient method, called the static method, has largely replaced the dynamic method, which uses a granular charcoal adsorbent. In the latter, the heated headspace is drawn over a short trap packed with charcoal, using a source of vacuum such as a pump or pushed along using pressurized nitrogen. The headspace volatiles in both the static and dynamic method are recovered by elution with a solvent, usually carbon disulfide. Recently, a promising variation of the static headspace method has been introduced. It is based on the use of a tiny amount of a polysiloxane polymer which has been coated onto the tip of a thin silica fiber. The fiber can be retracted into a syringe-type needle and the adsorbed headspace vapor can be conveniently introduced into the heated injector port of a gas chromatograph. No solvent is required. This technique, abbreviated SPME (for solid-phase microextraction) has many attractive advantages but it is not without some problems. Low boiling range accelerants, including water-soluble polar substances such as ethanol, are poorly retained on methylsiloxane type polymers. The recent

  13. A developed wedge fixtures assisted high precision TEM samples pre-thinning method: Towards the batch lamella preparation (United States)

    Wang, Dandan; Huang, Yamin; Liu, Binghai; Zhu, Lei; Lam, Jeffrey; Mai, Zhihong


    Ion milling, wedge cutting or polishing, and focused ion beam (FIB) milling are widely-used techniques for the transmission electron microscope (TEM) sample preparation. Especially, the FIB milling provides a site-specific analysis, deposition, and ablation of materials in the micrometer and nanometer scale. However, the cost of FIB tools has been always a significant concern. Since it is inevitable to use the FIB technique, the improvement of efficiency is a key point. Traditional TEM sample preparation with FIB was routinely implemented on a single sample each time. Aiming at cost efficiency, a new pre-thinning technique for batch sample preparation was developed in this paper. The present proposal combines the sample preparation techniques with multi-samples thinning, cross-section scanning electron microscopy (SEM), wedge cutting, FIB and other sample pre-thinning techniques. The new pre-thinning technique is to prepare an edge TEM sample on a grinding and polishing fixture with a slant surface. The thickness of the wedges sample can be measured to 1˜2 μm under optical microscope. Therefore, this fixture is superior to the traditional optical method of estimating the membrane thickness. Moreover, by utilizing a multi-sample holding fixture, more samples can be pre-thinned simultaneously, which significantly improved the productivity of TEM sample preparation.

  14. A developed wedge fixtures assisted high precision TEM samples pre-thinning method: Towards the batch lamella preparation

    Directory of Open Access Journals (Sweden)

    Dandan Wang


    Full Text Available Ion milling, wedge cutting or polishing, and focused ion beam (FIB milling are widely-used techniques for the transmission electron microscope (TEM sample preparation. Especially, the FIB milling provides a site-specific analysis, deposition, and ablation of materials in the micrometer and nanometer scale. However, the cost of FIB tools has been always a significant concern. Since it is inevitable to use the FIB technique, the improvement of efficiency is a key point. Traditional TEM sample preparation with FIB was routinely implemented on a single sample each time. Aiming at cost efficiency, a new pre-thinning technique for batch sample preparation was developed in this paper. The present proposal combines the sample preparation techniques with multi-samples thinning, cross-section scanning electron microscopy (SEM, wedge cutting, FIB and other sample pre-thinning techniques. The new pre-thinning technique is to prepare an edge TEM sample on a grinding and polishing fixture with a slant surface. The thickness of the wedges sample can be measured to 1∼2 μm under optical microscope. Therefore, this fixture is superior to the traditional optical method of estimating the membrane thickness. Moreover, by utilizing a multi-sample holding fixture, more samples can be pre-thinned simultaneously, which significantly improved the productivity of TEM sample preparation.

  15. Instrument and method for X-ray diffraction, fluorescence, and crystal texture analysis without sample preparation (United States)

    Gendreau, Keith (Inventor); Martins, Jose Vanderlei (Inventor); Arzoumanian, Zaven (Inventor)


    An X-ray diffraction and X-ray fluorescence instrument for analyzing samples having no sample preparation includes a X-ray source configured to output a collimated X-ray beam comprising a continuum spectrum of X-rays to a predetermined coordinate and a photon-counting X-ray imaging spectrometer disposed to receive X-rays output from an unprepared sample disposed at the predetermined coordinate upon exposure of the unprepared sample to the collimated X-ray beam. The X-ray source and the photon-counting X-ray imaging spectrometer are arranged in a reflection geometry relative to the predetermined coordinate.

  16. Improvement of a sample preparation method assisted by sodium deoxycholate for mass-spectrometry-based shotgun membrane proteomics. (United States)

    Lin, Yong; Lin, Haiyan; Liu, Zhonghua; Wang, Kunbo; Yan, Yujun


    In current shotgun-proteomics-based biological discovery, the identification of membrane proteins is a challenge. This is especially true for integral membrane proteins due to their highly hydrophobic nature and low abundance. Thus, much effort has been directed at sample preparation strategies such as use of detergents, chaotropes, and organic solvents. We previously described a sample preparation method for shotgun membrane proteomics, the sodium deoxycholate assisted method, which cleverly circumvents many of the challenges associated with traditional sample preparation methods. However, the method is associated with significant sample loss due to the slightly weaker extraction/solubilization ability of sodium deoxycholate when it is used at relatively low concentrations such as 1%. Hence, we present an enhanced sodium deoxycholate sample preparation strategy that first uses a high concentration of sodium deoxycholate (5%) to lyse membranes and extract/solubilize hydrophobic membrane proteins, and then dilutes the detergent to 1% for a more efficient digestion. We then applied the improved method to shotgun analysis of proteins from rat liver membrane enriched fraction. Compared with other representative sample preparation strategies including our previous sodium deoxycholate assisted method, the enhanced sodium deoxycholate method exhibited superior sensitivity, coverage, and reliability for the identification of membrane proteins particularly those with high hydrophobicity and/or multiple transmembrane domains. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. A high-throughput sample preparation method for cellular proteomics using 96-well filter plates. (United States)

    Switzar, Linda; van Angeren, Jordy; Pinkse, Martijn; Kool, Jeroen; Niessen, Wilfried M A


    A high-throughput sample preparation protocol based on the use of 96-well molecular weight cutoff (MWCO) filter plates was developed for shotgun proteomics of cell lysates. All sample preparation steps, including cell lysis, buffer exchange, protein denaturation, reduction, alkylation and proteolytic digestion are performed in a 96-well plate format, making the platform extremely well suited for processing large numbers of samples and directly compatible with functional assays for cellular proteomics. In addition, the usage of a single plate for all sample preparation steps following cell lysis reduces potential samples losses and allows for automation. The MWCO filter also enables sample concentration, thereby increasing the overall sensitivity, and implementation of washing steps involving organic solvents, for example, to remove cell membranes constituents. The optimized protocol allowed for higher throughput with improved sensitivity in terms of the number of identified cellular proteins when compared to an established protocol employing gel-filtration columns. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Gel-aided sample preparation (GASP)?A simplified method for gel-assisted proteomic sample generation from protein extracts and intact cells


    Fischer, Roman; Benedikt M Kessler


    We describe a ?gel-assisted? proteomic sample preparation method for MS analysis. Solubilized protein extracts or intact cells are copolymerized with acrylamide, facilitating denaturation, reduction, quantitative cysteine alkylation, and matrix formation. Gel-aided sample preparation has been optimized to be highly flexible, scalable, and to allow reproducible sample generation from 50 cells to milligrams of protein extracts. This methodology is fast, sensitive, easy-to-use on a wide range of...

  19. Universal nucleic acids sample preparation method for cells, spores and their mixture (United States)

    Bavykin, Sergei [Darien, IL


    The present invention relates to a method for extracting nucleic acids from biological samples. More specifically the invention relates to a universal method for extracting nucleic acids from unidentified biological samples. An advantage of the presently invented method is its ability to effectively and efficiently extract nucleic acids from a variety of different cell types including but not limited to prokaryotic or eukaryotic cells and/or recalcitrant organisms (i.e. spores). Unlike prior art methods which are focused on extracting nucleic acids from vegetative cell or spores, the present invention effectively extracts nucleic acids from spores, multiple cell types or mixtures thereof using a single method. Important that the invented method has demonstrated an ability to extract nucleic acids from spores and vegetative bacterial cells with similar levels effectiveness. The invented method employs a multi-step protocol which erodes the cell structure of the biological sample, isolates, labels, fragments nucleic acids and purifies labeled samples from the excess of dye.

  20. New Sample Preparation Method for Quantification of Phenolic Compounds of Tea (Camellia sinensis L. Kuntze: A Polyphenol Rich Plant

    Directory of Open Access Journals (Sweden)

    P. A. Nimal Punyasiri


    Full Text Available Chemical analysis of the Sri Lankan tea (Camellia sinensis, L. germplasm would immensely contribute to the success of the tea breeding programme. However, the polyphenols, particularly catechins (flavan-3-ols, are readily prone to oxidation in the conventional method of sample preparation. Therefore, optimization of the present sample preparation methodology for the profiling of metabolites is much important. Two sample preparation methodologies were compared, fresh leaves (as in the conventional procedures and freeze-dried leaves (a new procedure, for quantification of major metabolites by employing two cultivars, one is known to be high quality black tea and the other low quality black tea. The amounts of major metabolites such as catechins, caffeine, gallic acid, and theobromine, recorded in the new sampling procedure via freeze-dried leaves, were significantly higher than those recorded in the conventional sample preparation procedure. Additionally new method required less amount of leaf sample for analysis of major metabolites and facilitates storage of samples until analysis. The freeze-dried method would be useful for high throughput analysis of large number of samples in shorter period without chemical deterioration starting from the point of harvest until usage. Hence, this method is more suitable for metabolite profiling of tea as well as other phenol rich plants.

  1. An automation-assisted generic approach for biological sample preparation and LC-MS/MS method validation. (United States)

    Zhang, Jie; Wei, Shimin; Ayres, David W; Smith, Harold T; Tse, Francis L S


    Although it is well known that automation can provide significant improvement in the efficiency of biological sample preparation in quantitative LC-MS/MS analysis, it has not been widely implemented in bioanalytical laboratories throughout the industry. This can be attributed to the lack of a sound strategy and practical procedures in working with robotic liquid-handling systems. Several comprehensive automation assisted procedures for biological sample preparation and method validation were developed and qualified using two types of Hamilton Microlab liquid-handling robots. The procedures developed were generic, user-friendly and covered the majority of steps involved in routine sample preparation and method validation. Generic automation procedures were established as a practical approach to widely implement automation into the routine bioanalysis of samples in support of drug-development programs.

  2. Blood plasma sample preparation method to determine thyroid hormone-disrupting compounds in Effect-Directed Analysis

    NARCIS (Netherlands)

    Simon, E.; Bytingsvik, J.; Jonker, W.; Leonards, P.E.G.; de Boer, J.; Jenssen, B.M.; Lie, E.; Aars, J.; Hamers, T.H.M.; Lamoree, M.H.


    A sample preparation method combining solid-phase extraction (SPE) and liquid-liquid extraction (LLE) was developed to be used in Effect-Directed Analysis (EDA) of blood plasma. Until now such a method was not available. It can be used for extraction of a broad range of thyroid hormone


    Directory of Open Access Journals (Sweden)

    Maja Vrkljan


    Full Text Available The purpose of this study was to determine an optimal dissolution method for silicate rock samples for further analytical purposes. Analytical FAAS method of determining cobalt, chromium, copper, nickel, lead and zinc content in gabbro sample and geochemical standard AGV-1 has been applied for verification. Dissolution in mixtures of various inorganic acids has been tested, as well as Na2CO3 fusion technique. The results obtained by different methods have been compared and dissolution in the mixture of HNO3 + HF has been recommended as optimal.

  4. A method for the measurement of shielding effectiveness of planar samples requiring no sample edge preparation or contact


    Marvin, Andrew C.; Dawson, Linda; Flintoft, Ian Dand; Dawson, John F.


    A method is presented for the measurement of shielding effectiveness of planar materials with nonconducting surfaces such as carbon fiber composites. The method overcomes edge termination problems with such materials by absorbing edge-diffracted energy. A dynamic range of up to 100 dB has been demonstrated over a frequency range of 1-8.5 GHz, depending on the size of the sample under test. Comparison with ASTM D4935 and nested reverberation measurements of shielding effectiveness shows good a...

  5. Preparation of Samples for Leaf Architecture Studies, A Method for Mounting Cleared Leaves

    Directory of Open Access Journals (Sweden)

    Alejandra Vasco


    Full Text Available Premise of the study: Several recent waves of interest in leaf architecture have shown an expanding range of approaches and applications across a number of disciplines. Despite this increased interest, examination of existing archives of cleared and mounted leaves shows that current methods for mounting, in particular, yield unsatisfactory results and deterioration of samples over relatively short periods. Although techniques for clearing and staining leaves are numerous, published techniques for mounting leaves are scarce. Methods and Results: Here we present a complete protocol and recommendations for clearing, staining, and imaging leaves, and, most importantly, a method to permanently mount cleared leaves. Conclusions: The mounting protocol is faster than other methods, inexpensive, and straightforward; moreover, it yields clear and permanent samples that can easily be imaged, scanned, and stored. Specimens mounted with this method preserve well, with leaves that were mounted more than 35 years ago showing no signs of bubbling or discoloration.

  6. 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)


    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.

  7. Inverse supercritical fluid extraction as a sample preparation method for the analysis of the nanoparticle content in sunscreen agents. (United States)

    Müller, David; Cattaneo, Stefano; Meier, Florian; Welz, Roland; de Vries, Tjerk; Portugal-Cohen, Meital; Antonio, Diana C; Cascio, Claudia; Calzolai, Luigi; Gilliland, Douglas; de Mello, Andrew


    We demonstrate the use of inverse supercritical carbon dioxide (scCO2) extraction as a novel method of sample preparation for the analysis of complex nanoparticle-containing samples, in our case a model sunscreen agent with titanium dioxide nanoparticles. The sample was prepared for analysis in a simplified process using a lab scale supercritical fluid extraction system. The residual material was easily dispersed in an aqueous solution and analyzed by Asymmetrical Flow Field-Flow Fractionation (AF4) hyphenated with UV- and Multi-Angle Light Scattering detection. The obtained results allowed an unambiguous determination of the presence of nanoparticles within the sample, with almost no background from the matrix itself, and showed that the size distribution of the nanoparticles is essentially maintained. These results are especially relevant in view of recently introduced regulatory requirements concerning the labeling of nanoparticle-containing products. The novel sample preparation method is potentially applicable to commercial sunscreens or other emulsion-based cosmetic products and has important ecological advantages over currently used sample preparation techniques involving organic solvents. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Sample preparation method considerations for integrated transcriptomic and proteomic analysis of tumors. (United States)

    Bhat, Anupama Rajan; Gupta, Manoj Kumar; Krithivasan, Priya; Dhas, Kunal; Nair, Jayalakshmi; Reddy, Ram Bhupal; Sudheendra, Holalugunda Vittalamurthy; Chavan, Sandip; Vardhan, Harsha; Darsi, Sujatha; Balakrishnan, Lavanya; Katragadda, Shanmukh; Kekatpure, Vikram; Suresh, Amritha; Tata, Pramila; Panda, Binay; Kuriakose, Moni A; Sirdeshmukh, Ravi


    Sample processing protocols that enable compatible recovery of differentially expressed transcripts and proteins are necessary for integration of the multiomics data applied in the analysis of tumors. In this pilot study, we compared two different isolation methods for extracting RNA and protein from laryngopharyngeal tumor tissues and the corresponding adjacent normal sections. In Method 1, RNA and protein were isolated from a single tissue section sequentially and in Method 2, the extraction was carried out using two different sections and two independent and parallel protocols for RNA and protein. RNA and protein from both methods were subjected to RNA-seq and iTRAQ-based LC-MS/MS analysis, respectively. Analysis of data revealed that a higher number of differentially expressed transcripts and proteins were concordant in their regulation trends in Method 1 as compared to Method 2. Cross-method comparison of concordant entities revealed that RNA and protein extraction from the same tissue section (Method 1) recovered more concordant entities that are missed in the other extraction method (Method 2) indicating heterogeneity in distribution of these entities in different tissue sections. Method 1 could thus be the method of choice for integrated analysis of transcriptome and proteome data. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Sample preparation and biomass determination of SRF model mixture using cryogenic milling and the adapted balance method. (United States)

    Schnöller, Johannes; Aschenbrenner, Philipp; Hahn, Manuel; Fellner, Johann; Rechberger, Helmut


    The biogenic fraction of a simple solid recovered fuel (SRF) mixture (80 wt% printer paper/20 wt% high density polyethylene) is analyzed with the in-house developed adapted balance method (aBM). This fairly new approach is a combination of combustion elemental analysis (CHNS) and a data reconciliation algorithm based on successive linearisation for evaluation of the analysis results. This method shows a great potential as an alternative way to determine the biomass content in SRF. However, the employed analytical technique (CHNS elemental analysis) restricts the probed sample mass to low amounts in the range of a few hundred milligrams. This requires sample comminution to small grain sizes (sample size reduction. This paper presents a proof of principle investigation of the sample preparation and analysis of an SRF model mixture with the use of cryogenic impact milling (final sample comminution) and the adapted balance method (determination of biomass content). The so derived sample preparation methodology (cutting mills and cryogenic impact milling) shows a better performance in accuracy and precision for the determination of the biomass content than one solely based on cutting mills. The results for the determination of the biogenic fraction are within 1-5% of the data obtained by the reference methods, selective dissolution method (SDM) and (14)C-method ((14)C-M). Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Evaluation of micro-colorimetric lipid determination method with samples prepared using sonication and accelerated solvent extraction methods. (United States)

    Billa, Nanditha; Hubin-Barrows, Dylan; Lahren, Tylor; Burkhard, Lawrence P


    Two common laboratory extraction techniques were evaluated for routine use with the micro-colorimetric lipid determination method developed by Van Handel (1985) [2] and recently validated for small samples by Inouye and Lotufo (2006) [1]. With the accelerated solvent extraction method using chloroform:methanol solvent and the colorimetric lipid determination method, 28 of 30 samples had significant proportional bias (α=1%, determined using standard additions) and 1 of 30 samples had significant constant bias (α=1%, determined using Youden Blank measurements). With sonic extraction, 0 of 6 samples had significant proportional bias (α=1%) and 1 of 6 samples had significant constant bias (α=1%). These demonstrate that the accelerated solvent extraction method with chloroform:methanol solvent system creates an interference with the colorimetric assay method, and without accounting for the bias in the analysis, inaccurate measurements would be obtained. Published by Elsevier B.V.

  11. Sample preparation and biomass determination of SRF model mixture using cryogenic milling and the adapted balance method

    Energy Technology Data Exchange (ETDEWEB)

    Schnöller, Johannes, E-mail:; Aschenbrenner, Philipp; Hahn, Manuel; Fellner, Johann; Rechberger, Helmut


    Highlights: • An alternative sample comminution procedure for SRF is tested. • Proof of principle is shown on a SRF model mixture. • The biogenic content of the SRF is analyzed with the adapted balance method. • The novel method combines combustion analysis and a data reconciliation algorithm. • Factors for the variance of the analysis results are statistically quantified. - Abstract: The biogenic fraction of a simple solid recovered fuel (SRF) mixture (80 wt% printer paper/20 wt% high density polyethylene) is analyzed with the in-house developed adapted balance method (aBM). This fairly new approach is a combination of combustion elemental analysis (CHNS) and a data reconciliation algorithm based on successive linearisation for evaluation of the analysis results. This method shows a great potential as an alternative way to determine the biomass content in SRF. However, the employed analytical technique (CHNS elemental analysis) restricts the probed sample mass to low amounts in the range of a few hundred milligrams. This requires sample comminution to small grain sizes (<200 μm) to generate representative SRF specimen. This is not easily accomplished for certain material mixtures (e.g. SRF with rubber content) by conventional means of sample size reduction. This paper presents a proof of principle investigation of the sample preparation and analysis of an SRF model mixture with the use of cryogenic impact milling (final sample comminution) and the adapted balance method (determination of biomass content). The so derived sample preparation methodology (cutting mills and cryogenic impact milling) shows a better performance in accuracy and precision for the determination of the biomass content than one solely based on cutting mills. The results for the determination of the biogenic fraction are within 1–5% of the data obtained by the reference methods, selective dissolution method (SDM) and {sup 14}C-method ({sup 14}C-M)

  12. Thermal conductivity measurement of sintered Vibro-packed fuel. 1. Study on sample preparation method

    Energy Technology Data Exchange (ETDEWEB)

    Mizuno, Mineo; Kosaka, Yuji; Ogawa, Shinta [Nuclear Development Corp., Tokai, Ibaraki (Japan)


    An experimental study was carried out in order to grasp the sintering condition of the Vibro-packed fuel bed. A disc shaped bed of UO{sub 2} particles manufactured by the tumbling granulation method was sintered in constant load and temperature using a creep examination machine. The following results were obtained. 1) Sintering for about 2 hours and 30 minutes under the temperature of 1700degC and the compression load condition of 7 MPa generated 4.5 - 4.7% strain of the bed and about 40% neck ratio between particles. 2) Larger neck ratio was observed in larger sintering strain bed. This result implies the aimed neck ratio can be prepared by the adjustment of the sintering strain. 3) Sintering strain of the bed was depend on the particle size. In case of the large size particle, sintering strain became large. It was considered that it was based on the increase of local stress caused by the reduction of contact points according to particle diameter increase. 4) When the particle diameter becomes small, the particle neck ratio perpendicular to the load direction became small in comparison with the load direction, and the tendency that a particle becomes easy to separate was recognized. 5) >From the grain size measurement result of particle before and after a sintering experiment, no significant change of the crystal grain size was recognized. (author)

  13. Efficient sample preparation method based on solvent-assisted dispersive solid-phase extraction for the trace detection of butachlor in urine and waste water samples. (United States)

    Aladaghlo, Zolfaghar; Fakhari, Alireza; Behbahani, Mohammad


    In this work, an efficient sample preparation method termed solvent-assisted dispersive solid-phase extraction was applied. The used sample preparation method was based on the dispersion of the sorbent (benzophenone) into the aqueous sample to maximize the interaction surface. In this approach, the dispersion of the sorbent at a very low milligram level was achieved by inserting a solution of the sorbent and disperser solvent into the aqueous sample. The cloudy solution created from the dispersion of the sorbent in the bulk aqueous sample. After pre-concentration of the butachlor, the cloudy solution was centrifuged and butachlor in the sediment phase dissolved in ethanol and determined by gas chromatography with flame ionization detection. Under the optimized conditions (solution pH = 7.0, sorbent: benzophenone, 2%, disperser solvent: ethanol, 500 μL, centrifuged at 4000 rpm for 3 min), the method detection limit for butachlor was 2, 3 and 3 μg/L for distilled water, waste water, and urine sample, respectively. Furthermore, the preconcentration factor was 198.8, 175.0, and 174.2 in distilled water, waste water, and urine sample, respectively. Solvent-assisted dispersive solid-phase extraction was successfully used for the trace monitoring of butachlor in urine and waste water samples. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. A Microbiome DNA Enrichment Method for Next-Generation Sequencing Sample Preparation. (United States)

    Yigit, Erbay; Feehery, George R; Langhorst, Bradley W; Stewart, Fiona J; Dimalanta, Eileen T; Pradhan, Sriharsa; Slatko, Barton; Gardner, Andrew F; McFarland, James; Sumner, Christine; Davis, Theodore B


    "Microbiome" is used to describe the communities of microorganisms and their genes in a particular environment, including communities in association with a eukaryotic host or part of a host. One challenge in microbiome analysis concerns the presence of host DNA in samples. Removal of host DNA before sequencing results in greater sequence depth of the intended microbiome target population. This unit describes a novel method of microbial DNA enrichment in which methylated host DNA such as human genomic DNA is selectively bound and separated from microbial DNA before next-generation sequencing (NGS) library construction. This microbiome enrichment technique yields a higher fraction of microbial sequencing reads and improved read quality resulting in a reduced cost of downstream data generation and analysis. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.

  15. Benchmarking sample preparation/digestion protocols reveals tube-gel being a fast and repeatable method for quantitative proteomics. (United States)

    Muller, Leslie; Fornecker, Luc; Van Dorsselaer, Alain; Cianférani, Sarah; Carapito, Christine


    Sample preparation, typically by in-solution or in-gel approaches, has a strong influence on the accuracy and robustness of quantitative proteomics workflows. The major benefit of in-gel procedures is their compatibility with detergents (such as SDS) for protein solubilization. However, SDS-PAGE is a time-consuming approach. Tube-gel (TG) preparation circumvents this drawback as it involves directly trapping the sample in a polyacrylamide gel matrix without electrophoresis. We report here the first global label-free quantitative comparison between TG, stacking gel (SG), and basic liquid digestion (LD). A series of UPS1 standard mixtures (at 0.5, 1, 2.5, 5, 10, and 25 fmol) were spiked in a complex yeast lysate background. TG preparation allowed more yeast proteins to be identified than did the SG and LD approaches, with mean numbers of 1979, 1788, and 1323 proteins identified, respectively. Furthermore, the TG method proved equivalent to SG and superior to LD in terms of the repeatability of the subsequent experiments, with mean CV for yeast protein label-free quantifications of 7, 9, and 10%. Finally, known variant UPS1 proteins were successfully detected in the TG-prepared sample within a complex background with high sensitivity. All the data from this study are accessible on ProteomeXchange (PXD003841). © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. A comparison of sample preparation methods for extracting volatile organic compounds (VOCs) from equine faeces using HS-SPME. (United States)

    Hough, Rachael; Archer, Debra; Probert, Christopher


    Disturbance to the hindgut microbiota can be detrimental to equine health. Metabolomics provides a robust approach to studying the functional aspect of hindgut microorganisms. Sample preparation is an important step towards achieving optimal results in the later stages of analysis. The preparation of samples is unique depending on the technique employed and the sample matrix to be analysed. Gas chromatography mass spectrometry (GCMS) is one of the most widely used platforms for the study of metabolomics and until now an optimised method has not been developed for equine faeces. To compare a sample preparation method for extracting volatile organic compounds (VOCs) from equine faeces. Volatile organic compounds were determined by headspace solid phase microextraction gas chromatography mass spectrometry (HS-SPME-GCMS). Factors investigated were the mass of equine faeces, type of SPME fibre coating, vial volume and storage conditions. The resultant method was unique to those developed for other species. Aliquots of 1000 or 2000 mg in 10 ml or 20 ml SPME headspace were optimal. From those tested, the extraction of VOCs should ideally be performed using a divinylbenzene-carboxen-polydimethysiloxane (DVB-CAR-PDMS) SPME fibre. Storage of faeces for up to 12 months at - 80 °C shared a greater percentage of VOCs with a fresh sample than the equivalent stored at - 20 °C. An optimised method for extracting VOCs from equine faeces using HS-SPME-GCMS has been developed and will act as a standard to enable comparisons between studies. This work has also highlighted storage conditions as an important factor to consider in experimental design for faecal metabolomics studies.

  17. A validated HPLC-UV method and optimization of sample preparation technique for norepinephrine and serotonin in mouse brain. (United States)

    Thomas, Jaya; Khanam, Razia; Vohora, Divya


    Norepinephrine and serotonin are two important neurotransmitters whose variations in brain are reported to be associated with many common neuropsychiatric disorders. Yet, relevant literature on estimation of monoamines in biological samples using HPLC-UV is limited. The present study involves the development of a simultaneous HPLC-UV method for estimation of norepinephrine and serotonin along with optimization of the sample preparation technique. Chromatographic separation was achieved by injecting 20 µL of the sample after extraction into Quaternary pump HPLC equipped with C18 column using 0.05% formic acid and acetonitrile (90:10, v/v) as the mobile phase with 1 mL min(-1) flow rate. The developed method was validated as per the ICH guidelines in terms of linearity, accuracy, repeatability, precision, and robustness. The method showed a wide range of linearity (50-4000 and 31.25-4000 ng mL(-1) for norepinephrine and serotonin, respectively). The recovery was found to be in the range of 86.04-89.01% and 86.43-89.61% for norepinephrine and serotonin, respectively. The results showed low value of %RSD for repeatability, intra and inter-day precision, and robustness studies. Four different methods were used for the extraction of these neurotransmitters and the best one with maximum recovery was ascertained. Here, we developed and validated a simple, accurate, and reliable method for the estimation of norepinephrine and serotonin in mice brain samples using HPLC-UV. The method was successfully applied to quantify these neurotransmitters in mice brain extracted by optimized sample preparation technique.

  18. The choice of preparation method for the determination of Alternaria toxins from wheat samples by LC-MS/MS

    Directory of Open Access Journals (Sweden)

    Hajnal-Janić Elizabet P.


    Full Text Available Cereals are the primary source of human diet, wheat being the third most produced grain worldwide, and in Serbia second most produced grain, just behind corn. As a result of climate change and global warming, frequent occurrences of mycobiota on steep grains can produce a negative impact on the safety of food products and their quality, which inevitably leads to large economic losses. Although Fusarium spp. remains a main source of mycotoxins contamination of wheat, in recent years, due to the evident climatic changes affecting agricultural production, other mycotoxigenic fungi have been pointed out as important wheat contaminants. Among them are the fungi of the genus Alternaria, especially A. alternata, which under favourable conditions, produces mycotoxins such as alternariol, alternariol monomethyl ether, tenuazonic acid, and other Alternaria toxins. Taking into account the toxicity of metabolites produced by certain species of fungi of the genus Alternaria in the system from farm to table, it is necessary to develop specific and sensitive analytical methods in order to implement systematic controls of occurrence of Alternaria toxins. Liquid chromatography coupled to (tandem mass spectrometry (LC-MS/MS has become the technique of choice for the detection and quantification of Alternaria toxins in food and feed. There are several limiting factors such as the efficiency of sample cleanup and the lack of reference materials for food and feed. The aim of this study was to choose the most suitable preparation method for the determination of Alternaria toxins from wheat samples by LC-MS/MS technique based on published sample preparation methods, with possible modifications, which are used in analysis of mycotoxins. Modified method of wheat samples preparation by extraction with ethyl acetate was selected as acceptable based on extraction efficiency of analytes of interest.

  19. Comparative study of methods for DNA preparation from olive oil samples to identify cultivar SSR alleles in commercial oil samples: possible forensic applications. (United States)

    Breton, Catherine; Claux, Delphine; Metton, Isabelle; Skorski, Gilbert; Bervillé, André


    Virgin olive oil is made from diverse cultivars either mixed or single. Those ensure different tastes and typicity, and these may be also enhanced by the region of production of cultivars. The different olive oil labels correspond to their chemical composition and acidity. Labels also may correspond to a protected origin indication, and thus, such oils contain a given composition in cultivars. To verify the main cultivars used at the source of an olive oil sample, our method is based on DNA technology. DNA is present in all olive oil samples and even in refined oil, but the quantity may depend on the oil processing technology and oil conservation conditions. Thus, several supports were used to retain DNA checking different techniques (silica extraction, hydroxyapatite, magnetic beads, and spun column) to prepare DNA from variable amounts of oil. At this stage, it was usable for amplification through PCR technology and especially with the magnetic beads, and further purification processes were checked. Finally, the final method used magnetic beads. DNA is released from beads in a buffer. Once purified, we showed that it did not contain compounds inhibiting PCR amplification using SSR primers. Aliquot dilution fractions of this solution were successfully routinely used through PCR with different SSR primer sets. This enables confident detection of eventual alien alleles in oil samples. First applied to virgin oil samples of known composition, either single cultivars or mixtures of them, the method was verified working on commercial virgin oil samples using bottles bought in supermarkets. Last, we defined a protocol starting from 2 x 40 mL virgin olive oil, and DNA was prepared routinely in about 5 h. It was convenient to genotype together several loci per sample to check whether alleles were in accordance with those of expected cultivars. Thus, forensic applications of our method are expected. However, the method needs further improvement to work on all oil samples.

  20. A Green Analytical Method Using Ultrasound in Sample Preparation for the Flow Injection Determination of Iron, Manganese, and Zinc in Soluble Solid Samples by Flame Atomic Absorption Spectrometry

    Directory of Open Access Journals (Sweden)

    M. Carmen Yebra


    Full Text Available A simple and rapid analytical method was developed for the determination of iron, manganese, and zinc in soluble solid samples. The method is based on continuous ultrasonic water dissolution of the sample (5–30 mg at room temperature followed by flow injection flame atomic absorption spectrometric determination. A good precision of the whole procedure (1.2–4.6% and a sample throughput of ca. 25 samples h–1 were obtained. The proposed green analytical method has been successfully applied for the determination of iron, manganese, and zinc in soluble solid food samples (soluble cocoa and soluble coffee and pharmaceutical preparations (multivitamin tablets. The ranges of concentrations found were 21.4–25.61 μg g-1 for iron, 5.74–18.30 μg g-1 for manganese, and 33.27–57.90 μg g-1 for zinc in soluble solid food samples and 3.75–9.90 μg g-1 for iron, 0.47–5.05 μg g-1 for manganese, and 1.55–15.12 μg g-1 for zinc in multivitamin tablets. The accuracy of the proposed method was established by a comparison with the conventional wet acid digestion method using a paired t-test, indicating the absence of systematic errors.

  1. A Green Analytical Method Using Ultrasound in Sample Preparation for the Flow Injection Determination of Iron, Manganese, and Zinc in Soluble Solid Samples by Flame Atomic Absorption Spectrometry (United States)

    Yebra, M. Carmen


    A simple and rapid analytical method was developed for the determination of iron, manganese, and zinc in soluble solid samples. The method is based on continuous ultrasonic water dissolution of the sample (5–30 mg) at room temperature followed by flow injection flame atomic absorption spectrometric determination. A good precision of the whole procedure (1.2–4.6%) and a sample throughput of ca. 25 samples h–1 were obtained. The proposed green analytical method has been successfully applied for the determination of iron, manganese, and zinc in soluble solid food samples (soluble cocoa and soluble coffee) and pharmaceutical preparations (multivitamin tablets). The ranges of concentrations found were 21.4–25.61 μg g−1 for iron, 5.74–18.30 μg g−1 for manganese, and 33.27–57.90 μg g−1 for zinc in soluble solid food samples and 3.75–9.90 μg g−1 for iron, 0.47–5.05 μg g−1 for manganese, and 1.55–15.12 μg g−1 for zinc in multivitamin tablets. The accuracy of the proposed method was established by a comparison with the conventional wet acid digestion method using a paired t-test, indicating the absence of systematic errors. PMID:22567553

  2. Preparation and characterization of pyrromethene-567 dye-doped polymer samples using Gamma Irradiation Polymerization Method (GIPM)

    Energy Technology Data Exchange (ETDEWEB)

    Al-Ghamdi, Attieh A., E-mail: [Center of Nanotechnology, King Abdulaziz University, Jeddah (Saudi Arabia); Mahrous, Eiman M. [Taibah University, Department of Physics, Madinah (Saudi Arabia); Al-Enizi, Abdullah M. [King Saud University, Department of Chemistry, Riyadh (Saudi Arabia); Azam, Ameer [Center of Nanotechnology, King Abdulaziz University, Jeddah (Saudi Arabia)


    Dye-doped polymer gain media laser samples of pyrromethene-567 in a mixture with methyl methacrylate and 2-hydroxyethyl methacrylate copolymer have been prepared and synthesized by the Conventional Thermal Polymerization Method (CTPM) using an oven, and by the Gamma Irradiation Polymerization Method (GIPM) using Cobalt-60 ({sup 60}Co). Physical properties comprising absorption molar coefficients, absorption cross sections, fluorescence quantum yields, fluorescence life times, and emission cross sections were determined and calculated from the measured absorption and emission spectra. The efficiency and photo-stability of samples fabricated by both methods were measured and compared. The time required for synthesis was reduced by 90% using the GIPM. Complete polymerization of the GIPM sample was found to be at 7 kGy. Further, the GIPM produces high laser damage resistance material, which might be attributed to the highly dense polymeric network structure formed as a result of the effect of gamma radiation on MMA and HEMA monomers. - Highlights: • Samples of PM-567 in a mixture with MMA/HEMA were synthesized by the GIPM. • The time required for synthesis was reduced by 90% using the GIPM. • Complete polymerization of the GIPM sample was at 7 kGy. • The laser media produced is fast and free of contamination that gives high purity material. • The GIPM produced a high laser damage resistance material.

  3. Comparison of sample preparation methods for reliable plutonium and neptunium urinalysis using automatic extraction chromatography

    DEFF Research Database (Denmark)

    Qiao, Jixin; Xu, Yihong; Hou, Xiaolin


    ), it endows urinalysis methods with better reliability and repeatability compared with co-precipitation techniques. In view of the applicability of different pre-concentration techniques proposed previously in the literature, the main challenge behind relevant method development is pointed to be the release...

  4. Sample Preparation and Staining Methods for Two-Dimensional Polyacrylamide Gel Electrophoresis of Proteins from Animal Tissues

    Directory of Open Access Journals (Sweden)

    Levente Czegledi


    Full Text Available Proteomics in animal science as well as in other biological sciences is a significant tool in the post-genomic era. In proteomic studies the presence and relative abundance of expressed proteins of a cell, tissue or biological fluid is studied. Recently, the whole genome of more and more domestic animal species is known, but genes and the transcribed mRNA have no direct effect on biological systems as they are regulated by proteins, which explain the importance of proteomics. The most common tool in proteomic approach is the two-dimensional polyacrylamide gel electrophoresis (2D PAGE, when proteins are separated by their isoelectric point followed by their mass separation as a second dimension. In this study authors used different sample preparation and protein staining methods on meat,  liver and blood plasma and carried out 2D PAGE experiments. The most appropriate sample preparation methods are described in this paper. We concluded that depletion of major proteins in plasma is required but not necessary for meat and liver samples.

  5. Impacts of sample preparation methods on solubility and antilisterial characteristics of essential oil components in milk. (United States)

    Chen, Huaiqiong; Davidson, P Michael; Zhong, Qixin


    Essential oil components (EOCs) have limited water solubility and are used at much higher concentrations in complex food matrices than in growth media to inhibit pathogens. However, the correlation between solubility and activity has not been studied. The objective of this work was to characterize the solubility of EOCs in solvents and milk and correlate solubility with antilisterial activity. The solubilities of four EOCs, thymol, carvacrol, eugenol, and trans-cinnamaldehyde, in water was significantly increased in the presence of 5% (vol/vol) ethanol. In milk, the solubility of EOCs was lower than in water, with lower solubility in higher-fat milk. EOCs applied to milk as stock solutions (in 95% aqueous ethanol) enabled quicker dissolution and higher solubility in milk serum than other methods of mixing, such as end to end, and greater reductions of Listeria monocytogenes Scott A after 0 and 24 h. When the EOC concentration detected in milk serum was above the minimum bactericidal concentration, complete inhibition of L. monocytogenes in tryptic soy broth resulted. Therefore, the antilisterial properties in milk could be correlated with the solubility by comparison to the minimum inhibitory or bactericidal concentrations of EOCs. While the EOCs applied using ethanol generally had solubility and activity characteristics superior to those of other mixing methods, ethanol is not used to a great extent in nonfermented foods. Therefore, mixing methods without an organic solvent may be more readily adaptable to enhancing the distribution of EOCs in complex food systems.

  6. The beauty of being (label)-free: sample preparation methods for SWATH-MS and next-generation targeted proteomics. (United States)

    Vowinckel, Jakob; Capuano, Floriana; Campbell, Kate; Deery, Michael J; Lilley, Kathryn S; Ralser, Markus


    The combination of qualitative analysis with label-free quantification has greatly facilitated the throughput and flexibility of novel proteomic techniques. However, such methods rely heavily on robust and reproducible sample preparation procedures. Here, we benchmark a selection of in gel, on filter, and in solution digestion workflows for their application in label-free proteomics. Each procedure was associated with differing advantages and disadvantages. The in gel methods interrogated were cost effective, but were limited in throughput and digest efficiency. Filter-aided sample preparations facilitated reasonable processing times and yielded a balanced representation of membrane proteins, but led to a high signal variation in quantification experiments. Two in solution digest protocols, however, gave optimal performance for label-free proteomics. A protocol based on the detergent RapiGest led to the highest number of detected proteins at second-best signal stability, while a protocol based on acetonitrile-digestion, RapidACN, scored best in throughput and signal stability but came second in protein identification. In addition, we compared label-free data dependent (DDA) and data independent (SWATH) acquisition on a TripleTOF 5600 instrument. While largely similar in protein detection, SWATH outperformed DDA in quantification, reducing signal variation and markedly increasing the number of precisely quantified peptides.

  7. Sample preparation methods for the determination of plutonium and strontium in environmental samples by low level liquid scintillation counting and {alpha}-spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Solatie, D.; Carbol, P.; Hrnecek, E.; Betti, M. [European Commission, Joint Research Centre, Inst. for Transuranium Elements, Karlsruhe (Germany); Jaakkola, T. [Lab. of Radiochemistry, Univ. of Helsinki, Helsinki (Finland)


    Two different methods - leaching and microwave assisted total dissolution - have been exploited for the treatment of environmental samples for the determination of plutonium and strontium. Leaching applied to reference materials demonstrated the procedure to be applicable for the recovery of technogenic Pu and Sr from environmental samples. For the measurement of the alpha emitters of plutonium, co-precipitation with calcium oxalate and ferric hydroxide and separation with anion exchange has been used. For preparation of {alpha}-spectrometry sources, co-precipitation with NdF{sub 3} on a membrane filter or electro-deposition using the (NH{sub 4}){sub 2}C{sub 2}O{sub 4}/HCl method have been tested. The beta emitter {sup 241}Pu was measured by liquid scintillation counting. Pu isotope concentrations determined in the reference materials agreed well with the certified concentrations. {sup 90}Sr was measured in the leachate solutions from environmental samples collected close to a nuclear facility and from reference materials, after separation from the other leached elements, by liquid scintillation counting and Cherenkov counting. The {sup 90}Sr-concentrations determined in the reference materials agreed well with the certified concentrations. In the samples collected close a nuclear facility (soil, grass and sheep faeces), {sup 90}Sr was found at higher levels, which could also be correlated with the location of the sampling. (orig.)

  8. A method for sampling and tissue preparation of the parathyroid glands in miniature pigs for toxicity studies. (United States)

    Soshin, Tomomi; Takai, Hirotake; Kato, Chie; Fujii, Etsuko; Matsuo, Saori; Ito, Tsuneo; Suzuki, Masami


    Miniature swine (minipigs) are often used in non-rodent toxicity studies. However, unlike other animal species, the parathyroid glands of minipigs are often covered with thymic tissue and are similar in color, making macroscopical identification difficult. We investigated a method for sampling and tissue preparation of the parathyroid glands using 5- to 7-month-old minipigs. The glands were identified by finding the insertion site of a branch from the carotid artery into the cervical part of the thymus. Then the glands were marked and sampled. In a preliminary study, the glands were macroscopically and microscopically detected in 3/8 animals. The glands were not identified macroscopically in 5/8 animals but were detected in 3 of these animals. In total, we succeeded in detection of the glands in 6/8 animals (75%). The method was applied in the main toxicity study, and we succeeded in 100% detection through technical advancement. The method described herein enables high rate of detection and is useful in the pathological evaluation of the parathyroid glands of minipigs.

  9. A "three-in-one" sample preparation method for simultaneous determination of B-group water-soluble vitamins in infant formula using VitaFast(®) kits. (United States)

    Zhang, Heng; Lan, Fang; Shi, Yupeng; Wan, Zhi-Gang; Yue, Zhen-Feng; Fan, Fang; Lin, Yan-Kui; Tang, Mu-Jin; Lv, Jing-Zhang; Xiao, Tan; Yi, Changqing


    VitaFast(®) test kits designed for the microbiological assay in microtiter plate format can be applied to quantitative determination of B-group water-soluble vitamins such as vitamin B12, folic acid and biotin, et al. Compared to traditional microbiological methods, VitaFast(®) kits significantly reduce sample processing time and provide greater reliability, higher productivity and better accuracy. Recently, simultaneous determination of vitamin B12, folic acid and biotin in one sample is urgently required when evaluating the quality of infant formulae in our practical work. However, the present sample preparation protocols which are developed for individual test systems, are incompatible with simultaneous determination of several analytes. To solve this problem, a novel "three-in-one" sample preparation method is herein developed for simultaneous determination of B-group water-soluble vitamins using VitaFast(®) kits. The performance of this novel "three-in-one" sample preparation method was systematically evaluated through comparing with individual sample preparation protocols. The experimental results of the assays which employed "three-in-one" sample preparation method were in good agreement with those obtained from conventional VitaFast(®) extraction methods, indicating that the proposed "three-in-one" sample preparation method is applicable to the present three VitaFast(®) vitamin test systems, thus offering a promising alternative for the three independent sample preparation methods. The proposed new sample preparation method will significantly improve the efficiency of infant formulae inspection. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Sample preparation in biological mass spectrometry

    CERN Document Server

    Ivanov, Alexander R


    The aim of this book is to provide the researcher with important sample preparation strategies in a wide variety of analyte molecules, specimens, methods, and biological applications requiring mass spectrometric analysis as a detection end-point.

  11. A rapid HPLC column switching method for sample preparation and determination of β-carotene in food supplements. (United States)

    Brabcová, Ivana; Hlaváčková, Markéta; Satínský, Dalibor; Solich, Petr


    A simple and automated HPLC column-switching method with rapid sample pretreatment has been developed for quantitative determination of β-carotene in food supplements. Commercially samples of food supplements were dissolved in chloroform with help of saponification with 1M solution of sodium hydroxide in ultrasound bath. A 20-min sample dissolution/extraction step was necessary before chromatography analysis to transfer β-carotene from solid state of food supplements preparations (capsules,tablets) to chloroform solution. Sample volume - 3μL of chloroform phase was directly injected into the HPLC system. Next on-line sample clean-up was achieved on the pretreatment precolumn Chromolith Guard Cartridge RP-18e (Merck), 10×4.6mm, with a washing mobile phase (methanol:water, 92:8, (v/v)) at a flow rate of 1.5mL/min. Valve switch to analytical column was set at 2.5min in a back-flush mode. After column switching to the analytical column Ascentis Express C-18, 30×4.6mm, particle size 2.7μm (Sigma Aldrich), the separation and determination of β-carotene in food supplements was performed using a mobile phase consisting of 100% methanol, column temperature at 60°C and flow rate 1.5mL/min. The detector was set at 450nm. Under the optimum chromatographic conditions standard calibration curve was measured with good linearity - correlation coefficient for β-carotene (r(2)=0.999014; n=6) between the peak areas and concentration of β-carotene 20-200μg/mL. Accuracy of the method defined as a mean recovery was in the range 96.66-102.40%. The intraday method precision was satisfactory at three concentration levels 20, 125 and 200μg/mL and relative standard deviations were in the range 0.90-1.02%. The chromatography method has shown high sample throughput during column-switching pretreatment process and analysis in one step in short time (6min) of the whole chromatographic analysis. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR. (United States)

    Capote, Nieves; Bertolini, Edson; Olmos, Antonio; Vidal, Eduardo; Martínez, Maria Carmen; Cambra, Mariano


    Direct systems to process plant materials allowed high-throughput testing of Plum pox virus (PPV) by real-time reverse transcription (RT)-PCR without nucleic acids purification. Crude plant extracts were diluted in buffer or spotted on membranes to be used as templates. Alternatively, immobilized PPV targets were amplified from fresh sections of plant tissues printed or squashed onto the same supports, without extract preparation. Spot real-time RT-PCR was validated as a PPV diagnostic method in samples collected during the dormancy period and showed high sensitivity (93.6%), specificity (98.0%), and post-test probability (97.9%) towards sharka disease. In an analysis of 2919 Prunus samples by spot real-time RT-PCR and DASI-ELISA 90.8% of the results coincided, demonstrating high agreement (k = 0.77 +/- 0.01) between the two techniques. These results validate the use of immobilized PPV targets and spot real-time RT-PCR as screening method for largescale analyses.

  13. A fast and sensitive LC-MS/MS method for the quantification of fosfomycin in human urine and plasma using one sample preparation method and HILIC chromatography. (United States)

    Wijma, Rixt A; Bahmany, Soma; Wilms, E B; van Gelder, T; Mouton, Johan W; Koch, Birgit C P


    Fosfomycin is an old antibiotic that is increasingly prescribed because of emergence of the antibiotic resistance and the growing incidence of multi-drug resistant infections. Surprisingly, little is known about its pharmacokinetics (PK) and the pharmacodynamics (PD). Quantification of fosfomycin in both urine and plasma provides insight into the PK/PD characteristics of fosfomycin, which is crucial for the optimization of the therapy and the prevention of the emergence of resistance. An analytical method is therefore needed for the quantification of fosfomycin in both urine and plasma. A fast and sensitive tandem mass spectrometry method in combination with HILIC chromatography for the quantification of fosfomycin with a universal sample preparation method for urine and plasma was developed and validated according to FDA guidelines. The universal sample preparation method only requires 100μL of a sample, the addition of the internal standard fosfomycin-13C3 benzylamine and an ultrafiltration step. The method is applicable for the concentration range of 0.75-375mg/L (R2 of 0.9998 in both matrices) encompassing the clinically relevant concentration range based on the susceptibility of possible (uro)pathogens in the clinical setting. The validation results for urine and plasma for all QC levels, were samples were stable at 4°C and 15°C for at least 72h and stored samples at -80°C were stable for at least 6 months. Selectivity and sensitivity were confirmed and no carry-over was observed. The method was successfully applied in two pharmacokinetic studies in healthy volunteers and patients respectively. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Comparison of different methods for preparation and characterization of total RNA from cartilage samples to uncover osteoarthritis in vivo. (United States)

    Ruettger, Anke; Neumann, Steffi; Wiederanders, Bernd; Huber, René


    The isolation of intact RNA can be very difficult when tissues are used that contain many RNAses or that are hard to homogenize, e.g. cartilage samples. Additionally, cartilaginous tissues are characterized by a low cellularity and an abundance of extracellular matrix (ECM) molecules. But given the growing interest in understanding pathogenesis of degenerative diseases, e.g. osteoarthritis (OA) and rheumatoid arthritis (RA), studies have to consider expression pattern of cells in its natural environment. We compared the current RNA isolation methods for the extraction of high-quality RNA of snap-frozen biopsies from limited amounts of hypocellular cartilaginous tissue. The focus of the study was to gather information about procedure-related differences in RNA quality and yield. Here, we describe two protocols, the phenol/chloroform-free filter-based method (RNAqueous kit) and the combined protocol (TRIzol(R)/RNeasy Mini kit), working in a reproducible and reliable manner. We conclude that preparation, storage, homogenization, and quality control are altogether critical steps for in-depth analysis of differential gene expression, especially in hypocellular tissues with highly crosslinked ECM like cartilage.

  15. Comparison of different sample preparation methods for platinum determination in cultured cells by graphite furnace atomic absorption spectrometry

    Directory of Open Access Journals (Sweden)

    Man Xiao


    Full Text Available Background Platinum-based agents are widely used in chemotherapy against solid tumors and insufficient intracellular drug accumulation is one of the leading causes of platinum resistance which is associated with poor survival of tumor patients. Thus, the detection of intracellular platinum is pivotal for studies aiming to overcome platinum resistance. In the present study, we aimed to establish a reliable graphite furnace atomic absorption spectrometry (GFAAS-based assay to quantify the intracellular platinum content for cultured cells. Methods Several most commonly applied cell preparation methods, including 0.2% HNO3, 0.2% Triton X-100, concentrated nitric acid, RIPA combined with concentrated nitric acid and hydroxide, followed by GFAAS for platinum detection were compared in ovarian, cervical and liver cancer cell lines to obtain the optimal one, and parameters regarding linearity, accuracy, precision and sensitivity were evaluated. Influence of other metals on platinum detection and the storage conditions of samples were also determined. Results The treatment of cells with 0.2% HNO3 was superior to other approaches with fewer platinum loss and better repeatability. The recovery rate and precision of this method were 97.3%–103.0% and 1.4%–3.8%, respectively. The average recoveries in the presence of other metals were 95.1%–103.1%. The detection limit was 13.23 ug/L. The recovery rate of platinum remained acceptable even in cell samples stored in −20 °C or −80 °C for two months. Discussion After comparison, we found that 0.2% HNO3 was optimal for intracellular platinum quantification based on GFAAS, which presented values compatible with that of inductively-coupled plasma mass-spectrometry (ICP-MS, and this is partially attributed to the simplicity of this method. Moreover, the assay was proved to be accurate, sensitive, cost-effective and suitable for the research of platinum-based antitumor therapy.

  16. Sample-first preparation: a method for surface-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of cyclic oligosaccharides. (United States)

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


    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.

  17. Characterization of solid heterogeneous waste fuel - the effect of sampling and preparation method; Karaktaerisering av fasta inhomogena avfallsbraenslen - inverkan av metoder foer provtagning och provberedning

    Energy Technology Data Exchange (ETDEWEB)

    Wikstroem-Blomqvist, Evalena; Franke, Jolanta; Johansson, Ingvar


    The aim of the project is to evaluate the possibilities to simplify the methods used during sampling and laboratory preparation of heterogeneous waste materials. Existing methods for solid fuel material is summarized and evaluated in the project. As a result of the project two new simplified methods, one for field sampling and one for laboratory preparation work has been suggested. One large challenge regarding waste sampling is to achieve a representative sample due to the considerable heterogeneity of the material. How do you perform a sampling campaign that will give rise to representative results without too large costs? The single largest important source of error is the sampling procedure, equivalent to about 80% of the total error. Meanwhile the sample reduction and laboratory work only represents 15 % and 5 % respectively. Thus, to minimize the total error it is very important that the sampling is well planned in a testing program. In the end a very small analytical sample (1 gram) should reflected a large heterogeneous sample population of 1000 of tons. In this project two sampling campaigns, the fall of 2006 and early winter 2007, were conducted at the waste power plant Renova in Gothenburg, Sweden. The first campaign consisted of three different sample sizes with different number of sub-samples. One reference sample (50 tons and 48 sub-samples), two samples consisting of 16 tons and 8 sub-samples and finally two 4 tons consisting of 2 sub-samples each. During the second sampling campaign, four additional 4 ton samples were taken to repeat and thus evaluate the simplified sampling method. This project concludes that the simplified sampling methods only consisting of two sub-samples and a total sample volume of 4 tons give rise to results with as good quality and precision is the more complicated methods tested. Moreover the results from the two sampling campaigns generated equivalent results. The preparation methods used in the laboratory can as well be

  18. Microfluidic Sample Preparation for Immunoassays

    Energy Technology Data Exchange (ETDEWEB)

    Visuri, S; Benett, W; Bettencourt, K; Chang, J; Fisher, K; Hamilton, J; Krulevitch, P; Park, C; Stockton, C; Tarte, L; Wang, A; Wilson, T


    Researchers at Lawrence Livermore National Laboratory are developing means to collect and identify fluid-based biological pathogens in the forms of proteins, viruses, and bacteria. to support detection instruments, they are developing a flexible fluidic sample preparation unit. The overall goal of this Microfluidic Module is to input a fluid sample, containing background particulates and potentially target compounds, and deliver a processed sample for detection. They are developing techniques for sample purification, mixing, and filtration that would be useful to many applications including immunologic and nucleic acid assays. Many of these fluidic functions are accomplished with acoustic radiation pressure or dielectrophoresis. They are integrating these technologies into packaged systems with pumps and valves to control fluid flow through the fluidic circuit.

  19. An improved method of sample preparation on AnchorChip targets for MALDI-MS and MS/MS and its application in the liver proteome project

    DEFF Research Database (Denmark)

    Zhang, Xumin; Shi, Liang; Shu, Shaokung


    An improved method for sample preparation for MALDI-MS and MS/MS using AnchorChip targets is presented. The method, termed the SMW method (sample, matrix wash), results in better sensitivity for peptide mass fingerprinting as well as for sequencing by MS/MS than previously published methods. The ...... solution. The method was validated for protein identification from a 2-DE based liver proteome study. The SMW method resulted in identification of many more proteins and in most cases with a better score than the previously published methods....

  20. Trace element analysis of humus-rich natural water samples:method development for UV-LED assisted photocatalytic sample preparation and hydride generation ICP-MS analysis


    Havia, J. (Johanna)


    Abstract Humus-rich natural water samples, containing high concentrations of dissolved organic carbon (DOC), are challenging for certain analytical methods used in trace element analysis, including hydride generation methods and electrochemical methods. In order to obtain reliable results, the samples must to be pretreated to release analytes from humic acid complexes prior to the determination. In this study, methods for both pretreatment and analysis steps were developed. Arsenic is ...

  1. An efficient sample preparation method for high-throughput analysis of 15(S)-8-iso-PGF2α in plasma and urine by enzyme immunoassay. (United States)

    Bielecki, A; Saravanabhavan, G; Blais, E; Vincent, R; Kumarathasan, P


    Although several methods have been reported on the analysis of the oxidative stress marker 15(S)-8-iso-prostaglandin-F2alpha (8-iso-PGF2α) in biological fluids, they either involve extensive sample preparation and costly technology or require high sample volume. This study presents a sample preparation method that utilizes low sample volume for 8-iso-PGF2α analysis in plasma and urine by an enzyme immunoassay (EIA). In brief, 8-iso-PGF2α in deproteinized plasma or native urine sample is complexed with an antibody and then captured by molecular weight cut-off filtration. This method was compared with two other sample preparation methods that are typically used in the analysis of 8-iso-PGF2α by EIA: Cayman's affinity column purification method and solid-phase extraction on C-18. The immunoaffinity purification method described here was superior to the other two sample preparation methods and yielded recovery values of 99.8 and 54.1% for 8-iso-PGF2α in plasma and urine, respectively. Analytical precision (relative standard deviation) was ±5% for plasma and ±15% for urine. The analysis of healthy human plasma and urine resulted in basal 8-iso-PGF2α levels of 31.8 ± 5.5 pg/mL and 2.9 ± 2.0 ng/mg creatinine, respectively. The robustness and analytical performance of this method makes it a promising tool for high-throughput screening of biological samples for 8-iso-PGF2α.

  2. Urea free and more efficient sample preparation method for mass spectrometry based protein identification via combining the formic acid-assisted chemical cleavage and trypsin digestion. (United States)

    Wu, Shuaibin; Yang, Kaiguang; Liang, Zhen; Zhang, Lihua; Zhang, Yukui


    A formic acid (FA)-assisted sample preparation method was presented for protein identification via mass spectrometry (MS). Detailedly, an aqueous solution containing 2% FA and dithiothreitol was selected to perform protein denaturation, aspartic acid (D) sites cleavage and disulfide linkages reduction simultaneously at 108°C for 2h. Subsequently, FA wiped off via vacuum concentration. Finally, iodoacetamide (IAA) alkylation and trypsin digestion could be performed ordinally. A series of model proteins (BSA, β-lactoglobulin and apo-Transferrin) were treated respectively using such method, followed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) analysis. The identified peptide number was increased by ∼ 80% in comparison with the conventional urea-assisted sample preparation method. Moreover, BSA identification was achieved efficiently down to femtomole (25 ± 0 sequence coverage and 16 ± 1 peptides) via such method. In contrast, there were not peptides identified confidently via the urea-assisted method before desalination via the C18 zip tip. The absence of urea in this sample preparation method was an advantage for the more favorable digestion and MALDI-TOF MS analysis. The performances of two methods for the real sample (rat liver proteome) were also compared, followed by a nanoflow reversed-phase liquid chromatography with electrospray ionization tandem mass spectrometry system analysis. As a result, 1335 ± 43 peptides were identified confidently (false discovery rate method, corresponding to 295 ± 12 proteins (of top match=1 and requiring 2 unique peptides at least). In contrast, there were only 1107 ± 16 peptides (corresponding to 231 ± 10 proteins) obtained from the conventional urea-assisted method. It was serving as a more efficient protein sample preparation method for researching specific proteomes better, and providing assistance to develop other proteomics analysis methods, such as, peptide

  3. Modified Sample Preparation Approach for the Determination of the Phenolic and Humic-Like Substances in Natural Organic Materials By the Folin Ciocalteu Method. (United States)

    Pontoni, Ludovico; Panico, Antonio; Matanò, Alessia; van Hullebusch, Eric D; Fabbricino, Massimiliano; Esposito, Giovanni; Pirozzi, Francesco


    A novel modification of the sample preparation procedure for the Folin-Ciocalteu colorimetric assay for the determination of total phenolic compounds in natural solid and semisolid organic materials (e.g., foods, organic solid waste, soils, plant tissues, agricultural residues, manure) is proposed. In this method, the sample is prepared by adding sodium sulfate as a solid diluting agent before homogenization. The method allows for the determination of total phenols (TP) in samples with high solids contents, and it provides good accuracy and reproducibility. Additionally, this method permits analyses of significant amounts of sample, which reduces problems related to heterogeneity. We applied this method to phenols-rich lignocellulosic and humic-like solids and semisolid samples, including rice straw (RS), peat-rich soil (PS), and food waste (FW). The TP concentrations measured with the solid dilution (SD) preparation were substantially higher (increases of 41.4%, 15.5%, and 59.4% in RS, PS and FW, respectively) than those obtained with the traditional method (solids suspended in water). These results showed that the traditional method underestimates the phenolic contents in the studied solids.

  4. Evaluation of a fast and simple sample preparation method for PBDE flame retardants and DDT pesticides in fish for analysis by ELISA compared with GC-MS/MS (United States)

    A simple, fast, and cost-effective sample preparation method, previously developed and validated for the analysis of organic contaminants in fish using low-pressure gas chromatography tandem mass spectrometry (LPGC-MS/MS), was evaluated for analysis of polybrominated diphenyl ethers (PBDEs) and dich...

  5. Analysis of {sup 129}I in lichens by accelerator mass spectrometry through a microwave-based sample preparation method

    Energy Technology Data Exchange (ETDEWEB)

    Gomez-Guzman, J.M. [Centro Nacional de Aceleradores (CNA), Avda. Thomas Alva Edison 7, Isla de la Cartuja, 41092 Seville (Spain); Lopez-Gutierrez, J.M., E-mail: lguti@us.e [Centro Nacional de Aceleradores (CNA), Avda. Thomas Alva Edison 7, Isla de la Cartuja, 41092 Seville (Spain); Dpto. de Fisica Aplicada I, Escuela Universitaria Politecnica, c/. Virgen de Africa 7, 41011 Seville (Spain); Pinto, A.R. [Centro Nacional de Aceleradores (CNA), Avda. Thomas Alva Edison 7, Isla de la Cartuja, 41092 Seville (Spain); Holm, E. [Department of Radiation Physics, Lund University Hospital, S-22185 Lund (Sweden); Garcia-Leon, M. [Centro Nacional de Aceleradores (CNA), Avda. Thomas Alva Edison 7, Isla de la Cartuja, 41092 Seville (Spain); Dpto. Fisica Atomica, Molecular y Nuclear, Avda. Reina Mercedes, s/n, 41012 Seville (Spain)


    The presence of {sup 129}I in the environment has been strongly influenced by the artificial nuclear emissions since the beginning of the nuclear era in the mid 20th century. In order to know more about the different sources and their relative impact in different zones, it is necessary to complete the amount of measurements of this radionuclide in environmental samples. In this work, {sup 129}I has been determined in lichen samples (Cladonia alpestris) from Rogen Lake in Central Sweden. A method based on microwave digestion was developed for these measurements in order to improve speed and reduce contamination. Based on this method, {sup 129}I concentrations in some lichen samples from Lake Rogen (Sweden) have been measured, showing the impact of the Chernobyl accident and nuclear fuel reprocessing plants.

  6. Analysis of 129I in lichens by accelerator mass spectrometry through a microwave-based sample preparation method (United States)

    Gómez-Guzmán, J. M.; López-Gutiérrez, J. M.; Pinto, A. R.; Holm, E.; García-León, M.


    The presence of 129I in the environment has been strongly influenced by the artificial nuclear emissions since the beginning of the nuclear era in the mid 20th century. In order to know more about the different sources and their relative impact in different zones, it is necessary to complete the amount of measurements of this radionuclide in environmental samples. In this work, 129I has been determined in lichen samples ( Cladonia alpestris) from Rogen Lake in Central Sweden. A method based on microwave digestion was developed for these measurements in order to improve speed and reduce contamination. Based on this method, 129I concentrations in some lichen samples from Lake Rogen (Sweden) have been measured, showing the impact of the Chernobyl accident and nuclear fuel reprocessing plants.

  7. 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


    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...... protocol while being less labour intensive and more cost-effective due to minimal consumption of reagents, enzymes and consumables. Preliminary data obtained on a MALDI quadrupole-TOF tandem mass spectrometer demonstrated the utility of the on-probe digestion protocol for peptide mass mapping and peptide....../ionization-time of flight mass spectrometry (MALDI-TOF-MS) is used as the first protein screening method in many laboratories because of its inherent simplicity, mass accuracy, sensitivity and relatively high sample throughput. We present a simplified sample preparation method for MALDI-MS that enables in-gel digestion...

  8. Automated Sample Preparation (ASP): Development of a Rapid Method to Sequentially Isolate Nucleic Acids and Protein from Any Sample Type by a Cartridge-Based System (United States)


    by the buffer solution. For instance, the DNAPro appears to perform better when the diluent was Joint Portal Shield Buffer. This relationship did...spores and comparison of DNA yields from spores and spiked environmental samples. Journal of Microbiological Methods, 2009. 76(1): p. 30-37. 3...time PCR. European Journal of Clinical Microbiology & Infectious Diseases, 2008. 27(2): p. 109-114. 4. Boom, R., et al., Rapid and simple method

  9. Matrix removal in state of the art sample preparation methods for serum by charged aerosol detection and metabolomics-based LC-MS. (United States)

    Schimek, Denise; Francesconi, Kevin A; Mautner, Anton; Libiseller, Gunnar; Raml, Reingard; Magnes, Christoph


    Investigations into sample preparation procedures usually focus on analyte recovery with no information provided about the fate of other components of the sample (matrix). For many analyses, however, and particularly those using liquid chromatography-mass spectrometry (LC-MS), quantitative measurements are greatly influenced by sample matrix. Using the example of the drug amitriptyline and three of its metabolites in serum, we performed a comprehensive investigation of nine commonly used sample clean-up procedures in terms of their suitability for preparing serum samples. We were monitoring the undesired matrix compounds using a combination of charged aerosol detection (CAD), LC-CAD, and a metabolomics-based LC-MS/MS approach. In this way, we compared analyte recovery of protein precipitation-, liquid-liquid-, solid-phase- and hybrid solid-phase extraction methods. Although all methods provided acceptable recoveries, the highest recovery was obtained by protein precipitation with acetonitrile/formic acid (amitriptyline 113%, nortriptyline 92%, 10-hydroxyamitriptyline 89%, and amitriptyline N-oxide 96%). The quantification of matrix removal by LC-CAD showed that the solid phase extraction method (SPE) provided the lowest remaining matrix load (48-123 μg mL(-1)), which is a 10-40 fold better matrix clean-up than the precipitation- or hybrid solid phase extraction methods. The metabolomics profiles of eleven compound classes, comprising 70 matrix compounds showed the trends of compound class removal for each sample preparation strategy. The collective data set of analyte recovery, matrix removal and matrix compound profile was used to assess the effectiveness of each sample preparation method. The best performance in matrix clean-up and practical handling of small sample volumes was showed by the SPE techniques, particularly HLB SPE. CAD proved to be an effective tool for revealing the considerable differences between the sample preparation methods. This detector can

  10. Combination of dispersive liquid-liquid microextraction and solid-phase microextraction: An efficient hyphenated sample preparation method. (United States)

    Jafari, Mohammad T; Saraji, Mohammad; Mossaddegh, Mehdi


    Two well-known microextraction methods, dispersive liquid-liquid microextraction (DLLME) and solid-phase microextraction (SPME), were combined, resulting in as an encouraging method. The method, named DLLME-SPME, was performed based on total vaporization technique. For the DLLME step, 1,1,2,2-tetrachloroethane and acetonitrile were used as extraction and disperser solvents, respectively. Halloysite nanotubes-titanium dioxide was used as the fiber coating in the SPME step. The method was applied for the extraction of diazinon and parathion (as the test compounds) in environmental water samples and fruit juices, and gas chromatography-corona discharge ion mobility spectrometry was used as the determination apparatus. Desorption temperature and time, extraction temperature and time, and the volume of the extracting solvent in the DLLME step were optimized as the effective parameters on the extraction efficiency. The relative standard deviations (RSDs) of intra-day were found to be 4-7% and 6-8% for diazinon and parathion, respectively. Also, the RSDs of inter-day were 7-9% and 8-10% for diazinon and parathion, respectively. The limits of quantification and detection were obtained to be 0.015 and 0.005μgL(-1) for diazinon, and 0.020 and 0.007μgL(-1) for parathion. A good linearity range (r(2)˃0.993) was obtained in the range of 0.015-3.000 and 0.020-3.000μgL(-1) for diazinon and parathion, respectively. The high enrichment factors were obtained as 3150 and 2965 for diazinon and parathion, respectively. This method showed high sensitivity with good recovery values (between 87 and 99%) for the extraction of target analytes in the real samples. Overall, the results revealed that the developed DLLME-SPME method had better extraction efficiency than DLLME and SPME alone. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Green and efficient sample preparation method for the determination of catalyst residues in margarine by ICP-MS. (United States)

    Hartwig, Carla Andrade; Pereira, Rodrigo Mendes; Novo, Diogo La Rosa; Oliveira, Dirce Taina Teixeira; Mesko, Marcia Foster


    Responding to the need for green and efficient methods to determine catalyst residues with suitable precision and accuracy in samples with high fat content, the present work evaluates a microwave-assisted ultraviolet digestion (MW-UV) system for margarines and subsequent determination of Ni, Pd and Pt using inductively coupled plasma mass spectrometry (ICP-MS). It was possible to digest up to 500mg of margarine using only 10mL of 4molL -1 HNO 3 with a digestion efficiency higher than 98%. This allowed the determination of catalyst residues using the ICP-MS and free of interferences. For this purpose, the following experimental parameters were evaluated: concentration of digestion solution, sample mass and microwave irradiation program. The residual carbon content was used as a parameter to evaluate the efficiency of digestion and to select the most suitable experimental conditions. The accuracy evaluation was performed by recovery tests using a standard solution and certified reference material, and recoveries ranging from 94% to 99% were obtained for all analytes. The limits of detection for Ni, Pd and Pt using the proposed method were 35.6, 0.264 and 0.302ngg -1 , respectively. When compared to microwave-assisted digestion (MW-AD) in closed vessels using concentrated HNO 3 (used as a reference method for sample digestion), the proposed MW-UV could be considered an excellent alternative for the digestion of margarine, as this method requires only a diluted nitric acid solution for efficient digestion. In addition, MW-UV provides appropriate solutions for further ICP-MS determination with suitable precision (relative standard deviation catalyst residues was in agreement with the current legislation or recommendations. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. A new sample preparation method compatible with capillary electrophoresis and laser-induced fluorescence for improving detection of low levels of beta-lactoglobulin in infant foods. (United States)

    Pelaez-Lorenzo, Cristina; Diez-Masa, Jose Carlos; Vasallo, Isabel; de Frutos, Mercedes


    Beta-lactoglobulin (betaLG) is the main allergenic protein in cow's milk and can cause allergy even when present at very low concentration. The aim of this work is to develop an innovative sample preparation method fully compatible with capillary electrophoresis and laser-induced fluorescence detection for improving the sensitivity when analyzing betaLG. Different types of baby food were on purpose contaminated with diverse dairy desserts and submitted to thermal treatment to simulate potential contamination at production. Sample preparation prior to CE analysis was performed by the classical extraction method and by the innovative one, and the results were compared. Analysis was performed by capillary electrophoresis with laser-induced fluorescence detection. The innovative method permitted to detect contaminations as low as 1 part of yoghurt in 10,000 parts of baby food.

  13. Development of a Method for a Sensitive Simultaneous Determination of Dopamine and Paracetamol in Biological Samples and Pharmaceutical Preparations

    Directory of Open Access Journals (Sweden)

    Ali Babaei


    Full Text Available A chemically modified electrode is constructed based on multiwalled carbon nanotube—modified glassy carbon electrode (MWCNTs/GCE. The measurements were carried out by application of differential pulse voltammetry (DPV, cyclic voltammetry (CV, and chronoamperometry (CA methods. Application of DPV method showed wide linear range of DA from 1 μM to 540 μM and a detection limit of 0.098 μM (S/N=3. The linear range of PAR of 3 μM to 300 μM and a detection limit of 0.15 μM, were obtained. The modified electrode showed electrochemical responses with high sensitivity, high selectivity, and excellent stability for DA and PAR determination at optimal conditions, which makes it a suitable sensor for simultaneous submicromolar detection of DA and PAR in solutions. The analytical performance of this sensor has been evaluated for detection of DA and PAR in human serum, human urine, and pharmaceutical preparation with satisfactory results.

  14. Relevance of sample preparation for flow cytometry. (United States)

    Muccio, V E; Saraci, E; Gilestro, M; Oddolo, D; Ruggeri, M; Caltagirone, S; Bruno, B; Boccadoro, M; Omedè, P


    Flow cytometry is a useful tool for diagnosis and minimal residual disease (MRD) study of hematological diseases. Standard sample preparation protocols are characterized by stain-lyse-wash (SLW). To prevent nonspecific bindings and achieve high sensitivity in MRD studies, lyse-wash-stain-wash (LWSW) is required. To our knowledge, no comparison between the two methods has been performed. We compared mean fluorescence intensity (MFI), stain index, signal-to-noise ratio, and percentage of positive cells of 104 antibodies and of 13 selected antibodies tested in 10 samples simultaneously prepared with the two methods. MFI and percentages of positive cells obtained by the two methods did not show significant differences and showed a very high correlation. Stain index and signal-to-noise ratio presented higher values for kappa and lambda surface chains in LWSW samples and a trend of higher values for the other antibodies in SLW samples. We suggest to use LWSW method also at diagnosis to obtain more comparable antibody intensity expressions when samples from the same patient are processed for MRD evaluation after bulk lysis. Moreover, LWSW can prevent nonspecific bindings, shows no differences in the identification and quantitation of the populations of interest, and reduces acquisition of cell debris. © 2017 John Wiley & Sons Ltd.

  15. Comparison of pre-analytical FFPE sample preparation methods and their impact on massively parallel sequencing in routine diagnostics.

    Directory of Open Access Journals (Sweden)

    Carina Heydt

    Full Text Available Over the last years, massively parallel sequencing has rapidly evolved and has now transitioned into molecular pathology routine laboratories. It is an attractive platform for analysing multiple genes at the same time with very little input material. Therefore, the need for high quality DNA obtained from automated DNA extraction systems has increased, especially to those laboratories which are dealing with formalin-fixed paraffin-embedded (FFPE material and high sample throughput. This study evaluated five automated FFPE DNA extraction systems as well as five DNA quantification systems using the three most common techniques, UV spectrophotometry, fluorescent dye-based quantification and quantitative PCR, on 26 FFPE tissue samples. Additionally, the effects on downstream applications were analysed to find the most suitable pre-analytical methods for massively parallel sequencing in routine diagnostics. The results revealed that the Maxwell 16 from Promega (Mannheim, Germany seems to be the superior system for DNA extraction from FFPE material. The extracts had a 1.3-24.6-fold higher DNA concentration in comparison to the other extraction systems, a higher quality and were most suitable for downstream applications. The comparison of the five quantification methods showed intermethod variations but all methods could be used to estimate the right amount for PCR amplification and for massively parallel sequencing. Interestingly, the best results in massively parallel sequencing were obtained with a DNA input of 15 ng determined by the NanoDrop 2000c spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA. No difference could be detected in mutation analysis based on the results of the quantification methods. These findings emphasise, that it is particularly important to choose the most reliable and constant DNA extraction system, especially when using small biopsies and low elution volumes, and that all common DNA quantification techniques can


    Directory of Open Access Journals (Sweden)

    AVRAM Florin Timotei


    Full Text Available In this paper we propose to present the issues involved in the case of the constructive conception of a station for metallographic sample preparation. This station is destined for laboratory work. The metallographic station is composed of a robot ABB IRB1600, a metallographic microscope, a gripping device, a manipulator, a laboratory grinding and polishing machine. The robot will be used for manipulation of the sample preparation and the manipulator take the sample preparation for processing.

  17. Reversed-phase vortex-assisted liquid-liquid microextraction: a new sample preparation method for the determination of amygdalin in oil and kernel samples. (United States)

    Hosseini, Mohammad; Heydari, Rouhollah; Alimoradi, Mohammad


    A novel, simple, and rapid reversed-phase vortex-assisted liquid-liquid microextraction coupled with high-performance liquid chromatography has been introduced for the extraction, clean-up, and preconcentration of amygdalin in oil and kernel samples. In this technique, deionized water was used as the extracting solvent. Unlike the reversed-phase dispersive liquid-liquid microextraction, dispersive solvent was eliminated in the proposed method. Various parameters that affected the extraction efficiency, such as extracting solvent volume and its pH, vortex, and centrifuging times were evaluated and optimized. The calibration curve shows good linearity (r(2) = 0.9955) and precision (RSD < 5.2%) in the range of 0.07-20 μg/mL. The limit of detection and limit of quantitation were 0.02 and 0.07 μg/mL, respectively. The recoveries were in the range of 96.0-102.0% with relative standard deviation values ranging from 4.0 to 5.1%. Unlike the conventional extraction methods for plant extracts, no evaporative and re-solubilizing operations were needed in the proposed technique. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Solid Matrix Transformation and Tracer Addition using Molten Ammonium Bifluoride Salt as a Sample Preparation Method for Laser Ablation Inductively Coupled Plasma Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Grate, Jay W.; Gonzalez, Jhanis J.; O' Hara, Matthew J.; Kellogg, Cynthia M.; Morrison, Samuel S.; Koppenaal, David W.; Chan, George C.; Mao, Xianglei; Zorba, Vassilia; Russo, Richard


    Laser ablation (LA) is a means of sample introduction to inductively coupled plasma (ICP) mass spectrometry (MS) that avoids acid dissolution and chemical separation steps conventionally associated with solid sample analysis. At the same time, certain features of LA-ICP-MS are often mentioned in critical reviews including solid matrix variability and its influence on the ablation process, matrix dependent elemental fractionation, lack of matrix matched standards for external calibration, and limitations to internal calibration because it is challenging to add and distribute spikes into solid samples. In this paper we introduce the concept of a synergistic minimal sample preparation method that is used in combination with LA-ICP-MS as a means to overcome these limitations. The aim of this minimal sample preparation procedure is to reactively transform the original matrix to a more consistent matrix for LA-based analysis, thus reducing the effects of matrix variability, while enabling the addition of tracers. In conjunction with ICP-MS, we call this MTR-LA-ICP-MS, where MTR is derived from matrix transformation including the option to add tracers

  19. Shotgun analysis of membrane proteomes by an improved SDS-assisted sample preparation method coupled with liquid chromatography-tandem mass spectrometry. (United States)

    Lin, Yong; Jiang, Huajun; Yan, Yujun; Peng, Bin; Chen, Jinhua; Lin, Haiyan; Liu, Zhonghua


    Analysis of the membrane proteins, particularly the integral membrane proteins, is limited by the inherent membrane hydrophobicity. Sodium dodecyl sulfate (SDS) is one of the most efficient reagents used for the extraction of membrane proteins, but its presence in samples interferes with LC-MS-based proteomic analyses because it affects RP-LC separations and electrospray ionization. In this paper, we present an improved sample preparation strategy based on SDS-assisted digestion and peptide-level SDS-removal using an optimized potassium dodecyl sulfate (KDS) precipitation method (SSDP method) for shotgun analysis of the membrane proteome. This method utilizes a high concentration of SDS (1.0%) to lyse the membranes and to solubilize the hydrophobic membrane proteins, resulting in a more complete protein digestion in the diluted SDS buffer (0.1% SDS), and a high efficiency of SDS removal and peptide recovery by the optimized KDS precipitation for protein identification. The SSDP method provides evidence that proteins can be efficiently digested, and the SDS can be decreased to 95% peptide recovery. Compared to other sample preparation methods commonly used in shotgun membrane proteomics, the newly developed method not only increased the identified number of the total proteins, membrane proteins and integral membrane proteins by an average of 33.1%, 37.2% and 40.5%, respectively, but also leading to the identification of highest number of matching peptides. All the results showed that the method yielded better recovery and reliability in the identification of the proteins especially the highly hydrophobic integral membrane proteins, and thus providing a promising tool for the shotgun analysis of membrane proteome. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Finding even more anthropogenic indicators in mildly prepared sediment samples

    DEFF Research Database (Denmark)

    Enevold, Renée; Odgaard, Bent Vad


    NPPs in anthropogenic soils and archaeological samples are often numerous in types as well as in abundance. Preparing these soil samples with methods based on acid digestion holds the potential of severe bias leaving the NPP assemblages devoid of acid vulnerable NPPs. In many cases it might...... be worth the effort to prepare the NPP samples with as mild a preparation method as possible. We have mildly prepared NPP samples from a small forest hollow, Tårup Lund, Denmark. From the recovered NPP assemblages we attempt identifying anthropogenic indicators by comparing to the environmental information...

  1. Detection of Salmonella enterica in Meat in Less than 5 Hours by a Low-Cost and Noncomplex Sample Preparation Method. (United States)

    Fachmann, M S R; Löfström, C; Hoorfar, J; Hansen, F; Christensen, J; Mansdal, S; Josefsen, M H


    Salmonella is recognized as one of the most important foodborne bacteria and has wide health and socioeconomic impacts worldwide. Fresh pork meat is one of the main sources of Salmonella , and efficient and fast methods for detection are therefore necessary. Current methods for Salmonella detection in fresh meat usually include >16 h of culture enrichment, in a few cases meat, consisting of a 3-h enrichment in standard buffered peptone water and a real-time PCR-compatible sample preparation method based on filtration, centrifugation, and enzymatic digestion, followed by fast-cycling real-time PCR detection. The method was validated in an unpaired comparative study against the Nordic Committee on Food Analysis (NMKL) reference culture method 187. Pork meat samples ( n = 140) were either artificially contaminated with Salmonella at 0, 1 to 10, or 10 to 100 CFU/25 g of meat or naturally contaminated. Cohen's kappa for the degree of agreement between the rapid method and the reference was 0.64, and the relative accuracy, sensitivity, and specificity for the rapid method were 81.4, 95.1, and 97.9%, respectively. The 50% limit of detections (LOD 50 s) were 8.8 CFU/25 g for the rapid method and 7.7 CFU/25 g for the reference method. Implementation of this method will enable faster release of Salmonella low-risk meat, providing savings for meat producers, and it will help contribute to improved food safety. IMPORTANCE While the cost of analysis and hands-on time of the presented rapid method were comparable to those of reference culture methods, the fast product release by this method can provide the meat industry with a competitive advantage. Not only will the abattoirs save costs for work hours and cold storage, but consumers and retailers will also benefit from fresher meat with a longer shelf life. Furthermore, the presented sample preparation might be adjusted for application in the detection of other pathogenic bacteria in different sample types. Copyright © 2017

  2. New sample preparation method based on task-specific ionic liquids for extraction and determination of copper in urine and wastewater. (United States)

    Trtić-Petrović, Tatjana; Dimitrijević, Aleksandra; Zdolšek, Nikola; Đorđević, Jelena; Tot, Aleksandar; Vraneš, Milan; Gadžurić, Slobodan


    In this study, four hydrophilic ionic liquids (ILs) containing 1-alkyl-3-methylimidazolim cation and either salicylate or chloride anions were synthetized and studied as new task-specific ionic liquids (TSILs) suitable for aqueous biphasic system (ABS) formation and selective one-step extraction of copper(II). TSILs are designed so that the anion is responsible for forming the complex with metal(II) and preventing hydrolysis of metal cations at very strong alkaline pH, whereas the cation is responsible for selective extraction of metal(II)-salicylate complexes. It was found that 1-butyl-3-methylimidazolium salicylate could be used for selective extraction of Cu(II) in the presence of Zn(II), Cd(II), and Pb(II) at very alkaline solution without metal hydroxide formation. It was assumed that formation of metal(II)-salicylate complexes prevents the hydrolysis of the metal ions in alkaline solutions. The determined stability constants for Cu(II)-salicylate complexes, where salicylate was derived from different ionic liquids, indicated that there was no significant influence of the cation of the ionic liquid on the stability of the complexes. The ABS based on 1-butyl-3-methylimidazolium salicylate has been applied as the sample preparation step prior to voltammetric determination of Cu(II). The effect of volume of aqueous sample and IL and extraction time were investigated and optimum extraction conditions were determined. The obtained detection limits were 8 ng dm-3. The optimized method was applied for the determination of Cu(II) in tap water, wastewater, and urine. The study indicated that application of the ABS based on 1-butyl-3-methylimidazolium salicylate ionic liquid could be successfully applied as the sample preparation method for the determination of Cu(II) from various environmental samples. Graphical abstract Aqueous biphasic system based on task-specific ionic liquid as a sample pretreatment for selective determination of Cu(II) in biological and

  3. Diagnostic performance of a newly developed salivary cortisol and cortisone measurement using an LC-MS/MS method with simple and rapid sample preparation. (United States)

    Mészáros, K; Karvaly, G; Márta, Z; Magda, B; Tőke, J; Szücs, N; Tóth, M; Rácz, K; Patócs, A


    Late-night salivary cortisol level is one of the first-line tests recommended by the Endocrine Society for the diagnosis of endogenous hypercortisolism. Most routine laboratories measure cortisol levels using immunoassay tests which fail to determine low cortisol levels accurately due to the numerous interfering substances. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with simple and rapid sample preparation was developed for the simultaneous measurement of cortisol and cortisone and its performance in the diagnosis of endogenous hypercortisolism was evaluated. 324 late-night saliva samples were analyzed from which 272 samples were derived from patients with a suspected diagnosis of endogenous hypercortisolism. Salivary cortisol levels were assayed using an electrochemiluminescent immunoassay (ECLIA, Cortisol II, Roche), and simultaneous measurement of cortisol and cortisone was performed using an LC-MS/MS method. A strong correlation between cortisol results measured using ECLIA and LC-MS/MS (r 2 = 0.892) was demonstrated. Receiver operating characteristics (ROC) analysis showed good diagnostic performance of cortisol and cortisone levels assayed using LC-MS/MS method and for cortisol measured using ECLIA. Late-night salivary cortisol and cortisone are useful parameters for the diagnosis of hypercortisolism. Using samples obtained from patients where the diagnosis of hypercortisolism is extremely challenging cut-off values for midnight salivary cortisol and cortisone measured by LC-MS/MS method were established.

  4. Multiresidue method for the determination of 77 pesticides in wine using QuEChERS sample preparation and gas chromatography with mass spectrometry. (United States)

    Jiang, Y; Li, X; Xu, J; Pan, C; Zhang, J; Niu, W


    A method based on a QuEChERS (quick, easy, cheap, effective, rugged, safe) sample preparation method and gas chromatography with mass spectrometric detection by selected ion monitoring (GC/MS-SIM) was developed for simultaneous determination of 77 pesticide residues in wine. An extraction of 10 ml of sample with acetonitrile followed by liquid-liquid partition formed by the addition of 4 g MgSO(4) and 3 g NaCl was applied in the sample preparation. The clean-up was carried out by applying dispersive solid-phase with 150 mg MgSO(4) as well as 50 mg primary secondary amine (PSA). One quantitation ion and at least two identification ions were selected in the analytical method for each pesticide compound by GC/MS. The recovery data were obtained by spiking blank samples at two concentration levels (0.05 and 0.2 mg l(-1)). The recoveries of all pesticides were in the range 70-110%, with intra-day precision of less than 15%, and the inter-day precision of less than 22% and 15% for 0.05 and 0.2 mg l(-1) fortification levels, respectively. Linearity was between 0.02 and 2 mg l(-1) with determination coefficients (R(2)) greater than 0.98 for all compounds. The limits of quantification (LOQs) for the 77 pesticides ranged from 0.003 to 0.05 mg l(-1). This method was applied for routine analysis in market products.

  5. Sampling system and method (United States)

    Decker, David L.; Lyles, Brad F.; Purcell, Richard G.; Hershey, Ronald Lee


    The present disclosure provides an apparatus and method for coupling conduit segments together. A first pump obtains a sample and transmits it through a first conduit to a reservoir accessible by a second pump. The second pump further conducts the sample from the reservoir through a second conduit.

  6. Sample Preparation and Extraction in Small Sample Volumes Suitable for Pediatric Clinical Studies: Challenges, Advances, and Experiences of a Bioanalytical HPLC-MS/MS Method Validation Using Enalapril and Enalaprilat

    Directory of Open Access Journals (Sweden)

    Bjoern B. Burckhardt


    Full Text Available In USA and Europe, medicines agencies force the development of child-appropriate medications and intend to increase the availability of information on the pediatric use. This asks for bioanalytical methods which are able to deal with small sample volumes as the trial-related blood lost is very restricted in children. Broadly used HPLC-MS/MS, being able to cope with small volumes, is susceptible to matrix effects. The latter restrains the precise drug quantification through, for example, causing signal suppression. Sophisticated sample preparation and purification utilizing solid-phase extraction was applied to reduce and control matrix effects. A scale-up from vacuum manifold to positive pressure manifold was conducted to meet the demands of high-throughput within a clinical setting. Faced challenges, advances, and experiences in solid-phase extraction are exemplarily presented on the basis of the bioanalytical method development and validation of low-volume samples (50 μL serum. Enalapril, enalaprilat, and benazepril served as sample drugs. The applied sample preparation and extraction successfully reduced the absolute and relative matrix effect to comply with international guidelines. Recoveries ranged from 77 to 104% for enalapril and from 93 to 118% for enalaprilat. The bioanalytical method comprising sample extraction by solid-phase extraction was fully validated according to FDA and EMA bioanalytical guidelines and was used in a Phase I study in 24 volunteers.

  7. Sample Preparation and Extraction in Small Sample Volumes Suitable for Pediatric Clinical Studies: Challenges, Advances, and Experiences of a Bioanalytical HPLC-MS/MS Method Validation Using Enalapril and Enalaprilat. (United States)

    Burckhardt, Bjoern B; Laeer, Stephanie


    In USA and Europe, medicines agencies force the development of child-appropriate medications and intend to increase the availability of information on the pediatric use. This asks for bioanalytical methods which are able to deal with small sample volumes as the trial-related blood lost is very restricted in children. Broadly used HPLC-MS/MS, being able to cope with small volumes, is susceptible to matrix effects. The latter restrains the precise drug quantification through, for example, causing signal suppression. Sophisticated sample preparation and purification utilizing solid-phase extraction was applied to reduce and control matrix effects. A scale-up from vacuum manifold to positive pressure manifold was conducted to meet the demands of high-throughput within a clinical setting. Faced challenges, advances, and experiences in solid-phase extraction are exemplarily presented on the basis of the bioanalytical method development and validation of low-volume samples (50 μL serum). Enalapril, enalaprilat, and benazepril served as sample drugs. The applied sample preparation and extraction successfully reduced the absolute and relative matrix effect to comply with international guidelines. Recoveries ranged from 77 to 104% for enalapril and from 93 to 118% for enalaprilat. The bioanalytical method comprising sample extraction by solid-phase extraction was fully validated according to FDA and EMA bioanalytical guidelines and was used in a Phase I study in 24 volunteers.

  8. Free Amino Acid Profiles from 'Pinot Noir' Grapes are Influenced by Vine N-status and Sample Preparation Method (United States)

    This study examined the impact of extraction method on ammonia, free amino acids, and YAN (yeast assimilable nitrogen) concentrations in 'Pinot noir' berries obtained from a vine nutrition study (altered supply of N, P, or K). Berries were either juiced or exhaustively extracted as whole berries pri...

  9. Liquid-phase sample preparation method for real-time monitoring of airborne asbestos fibers by dual-mode high-throughput microscopy. (United States)

    Cho, Myoung-Ock; Kim, Jung Kyung; Han, Hwataik; Lee, Jeonghoon


    Asbestos that had been used widely as a construction material is a first-level carcinogen recognized by the World Health Organization. It can be accumulated in body by inhalation causing virulent respiratory diseases including lung cancer. In our previous study, we developed a high-throughput microscopy (HTM) system that can minimize human intervention accompanied by the conventional phase contrast microscopy (PCM) through automated counting of fibrous materials and thus significantly reduce analysis time and labor. Also, we attempted selective detection of chrysotile using DksA protein extracted from Escherichia coli through a recombinant protein production technique, and developed a dual-mode HTM (DM-HTM) by upgrading the HTM device. We demonstrated that fluorescently-labeled chrysotile asbestos fibers can be identified and enumerated automatically among other types of asbestos fibers or non-asbestos particles in a high-throughput manner through a newly modified HTM system for both reflection and fluorescence imaging. However there is a limitation to apply DM-HTM to airborne sample with current air collecting method due to the difficulty of applying the protein to dried asbestos sample. Here, we developed a technique for preparing liquid-phase asbestos sample using an impinger normally used to collect odor molecules in the air. It would be possible to improve the feasibility of the dual-mode HTM by integrating a sample preparation unit for making collected asbestos sample dispersed in a solution. The new technique developed for highly sensitive and automated asbestos detection can be a potential alternative to the conventional manual counting method, and it may be applied on site as a fast and reliable environmental monitoring tool.

  10. Method of applying sanitizers and sample preparation affects recovery of native microflora and Salmonella on whole cantaloupe surfaces. (United States)

    Ukuku, Dike O; Fett, William F


    Standardized methods for applying sanitizer treatments to cantaloupes and for recovering surviving native microflora or Salmonella on inoculated cantaloupe after sanitizing are lacking. Accordingly, the objectives of this study were to compare four methods for applying sanitizers (dipping, dipping with rotation, dipping with agitation, and dipping with rubbing) using 200 ppm of chlorine or 5% H2O2, two recovery methods (homogenization of rind plugs in a stomacher or blender), and five selective recovery media for Salmonella. Whole cantaloupes were submerged in a cocktail of five strains of Salmonella (each at approximately 2 x 10(8) CFU/ml) for 10 min and allowed to dry for 1 h inside a biosafety cabinet and stored at 20 degrees C for approximately 23 h before sanitizing. The recovery of Salmonella from whole cantaloupe without sanitizing averaged 5.09 log CFU/cm2 by blending and 4.30 log CFU/cm2 by homogenization in a stomacher for the five selective agar media. Microbial populations (Salmonella or the indigenous aerobic mesophilic bacteria, gram-negative bacteria, lactic acid bacteria, Pseudomonas spp., and yeast and mold) were not significantly (P > 0.05) reduced by treating with water regardless of the treatment method used. Sanitizing with chlorine or H2O2 by dipping, with or without rotation for 2 min, also did not reduce microbial populations. However, populations of all classes of native microflora and Salmonella were significantly (P sanitizer treatments (2 min) applied with agitation or by rubbing. In general, sanitizer treatments applied by rubbing resulted in greater log reductions (by up to 1.7 log unit) than for treatments applied with agitation. Populations of native microflora and Salmonella recovered from cantaloupe were higher (by up to 1.8 log unit) by blending compared to homogenization in a stomacher. In most instances, selective media used did not differ significantly (P > 0.05) for recovery of Salmonella after washing treatments.

  11. Evaluation of sample preparation methods for the detection of total metal content using inductively coupled plasma optical emission spectrometry (ICP-OES) in wastewater and sludge (United States)

    Dimpe, K. M.; Ngila, J. C.; Mabuba, N.; Nomngongo, P. N.

    Heavy metal contamination exists in aqueous wastes and sludge of many industrial discharges and domestic wastewater, among other sources. Determination of metals in the wastewater and sludge requires sample pre-treatment prior to analysis because of certain challenges such as the complexity of the physical state of the sample, which may lead to wrong readings in the measurement. This is particularly the case with low analyte concentration to be detected by the instrument. The purpose of this work was to assess and validate the different sample preparation methods namely, hot plate and microwave-assisted digestion procedures for extraction of metal ions in wastewater and sludge samples prior to their inductively coupled plasma optical emission spectrometric (ICP-OES) determination. For the extraction of As, Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Zn, three acid mixtures, that is, HNO3/H2O2, HNO3/HClO4/H2O2 and aqua regia + H2O2, were evaluated. Influent wastewater spiked with the SRM (CWW-TM-B) was used for the optimization of acid mixtures affecting the extraction procedure. After sample digestion, the filtration capabilities of cellulose-acetate filter paper and the acrodisc syringe filter with the pore size of 0.45 μm were compared. In terms of performance, acrodisc syringe filter in terms of the improved recoveries obtained, was found to be the best filtration method compared to the filter paper. Based on the analytical results obtained, microwave-assisted digestion (MAD) using aqua regia + H2O2 mixture was found to be the most suitable method for extraction of heavy metals and major elements in all the sample matrices. Therefore, MAD using aqua regia + H2O2 mixture was used for further investigations. The precision of the developed MAD method expressed in terms of relative standard deviations (% RSD) for different metals was found to be wastewater and sludge.

  12. A review of novel strategies of sample preparation for the determination of antibacterial residues in foodstuffs using liquid chromatography-based analytical methods

    Energy Technology Data Exchange (ETDEWEB)

    Marazuela, M.D., E-mail: [Department of Analytical Chemistry, Faculty of Chemistry, Universidad Complutense de Madrid, E-28040 Madrid (Spain); Bogialli, S. [Department of Chemistry, University of Rome ' La Sapienza' , Piazza Aldo Moro, 5 00185 Rome (Italy)


    The determination of trace residues and contaminants in food has been of growing concern over the past few years. Residual antibacterials in food constitute a risk to human health, especially because they can contribute to the transmission of antibiotic-resistant pathogenic bacteria through the food chain. Therefore, to ensure food safety EU and USA regulatory agencies have established lists of forbidden or banned substances and tolerance levels for authorized veterinary drugs (e.g. antibacterials). In addition, the EU Commission Decision 2002/657/EC has set requirements about the performance of analytical methods for the determination of veterinary drug residues in food and feedstuffs. During the past years, the use of powerful mass spectrometric detectors in combination with innovative chromatographic technologies has solved many problems related to sensitivity and selectivity of this type of analysis. However sample preparation still remains as the bottleneck step, mainly in terms of analysis time and sources of error. This review covering research published between 2004 and 2008 intends to provide an update overview of the past five years, on recent trends in sample preparation for the determination of antibacterial residues in foods, making special emphasis in on-line, high-throughput, multi-class methods and including several applications in detail.

  13. Preparation of Homogeneous MALDI Samples for Quantitative Applications. (United States)

    Ou, Yu-Meng; Tsao, Chien-Wei; Lai, Yin-Hung; Lee, Hsun; Chang, Huan-Tsung; Wang, Yi-Sheng


    This protocol demonstrates a simple sample preparation to reduce spatial heterogeneity in ion signals during matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The heterogeneity of ion signals is a severe problem in MALDI, which results in poor data reproducibility and makes MALDI unsuitable for quantitative analysis. By regulating sample plate temperature during sample preparation, thermal-induced hydrodynamic flows inside droplets of sample solution are able to reduce the heterogeneity problem. A room-temperature sample preparation chamber equipped with a temperature-regulated copper base block that holds MALDI sample plates facilitates precise control of the sample drying condition. After drying of sample droplets, the temperature of sample plates is returned to room temperature and removed from the chamber for subsequent mass spectrometric analysis. The areas of samples are examined with MALDI-imaging mass spectrometry to obtain the spatial distribution of all components in the sample. In comparison with the conventional dried-droplet method that prepares samples under ambient conditions without temperature control, the samples prepared with the method demonstrated herein show significantly better spatial distribution and signal intensity. According to observations using carbohydrate and peptide samples, decreasing substrate temperature while maintaining the surroundings at ambient temperature during the drying process can effectively reduce the heterogeneity of ion signals. This method is generally applicable to various combinations of samples and matrices.

  14. Quantification of piperacillin, tazobactam, cefepime, meropenem, ciprofloxacin and linezolid in serum using an isotope dilution UHPLC-MS/MS method with semi-automated sample preparation. (United States)

    Zander, Johannes; Maier, Barbara; Suhr, Anna; Zoller, Michael; Frey, Lorenz; Teupser, Daniel; Vogeser, Michael


    Recent studies have demonstrated highly variable blood concentrations of piperacillin, tazobactam, cefepime, meropenem, ciprofloxacin and linezolid in critically ill patients with a high incidence of sub-therapeutic levels. Consequently, therapeutic drug monitoring (TDM) of these antibiotics has to be considered, requiring robust and reliable routine analytical methods. The aim of the present work was to develop and validate a multi-analyte ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous quantification of the above mentioned antibiotics. Sample preparation included a manual protein precipitation step followed by two-dimensional ultra high performance liquid chromatography (2D-UHPLC). Corresponding stable isotope-labeled substances were used as internal standards for all of the analytes, with the exception of tazobactam. The injected sample volume was 7 μL. The run time was 5.0 min. Inaccuracy was ≤8% and imprecision coefficient of variation (CV) was method was found to be robust during the validation period. We were able to develop a reliable 2D-UHPLC-MS/MS method addressing analytes with highly heterogeneous physico-chemical properties. The novel assay may be an efficient tool for an optimized process workflow in clinical laboratories for important antibiotics in regards to TDM.

  15. Solid matrix transformation and tracer addition using molten ammonium bifluoride salt as a sample preparation method for laser ablation inductively coupled plasma mass spectrometry. (United States)

    Grate, Jay W; Gonzalez, Jhanis J; O'Hara, Matthew J; Kellogg, Cynthia M; Morrison, Samuel S; Koppenaal, David W; Chan, George C-Y; Mao, Xianglei; Zorba, Vassilia; Russo, Richard E


    Solid sampling and analysis methods, such as laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), are challenged by matrix effects and calibration difficulties. Matrix-matched standards for external calibration are seldom available and it is difficult to distribute spikes evenly into a solid matrix as internal standards. While isotopic ratios of the same element can be measured to high precision, matrix-dependent effects in the sampling and analysis process frustrate accurate quantification and elemental ratio determinations. Here we introduce a potentially general solid matrix transformation approach entailing chemical reactions in molten ammonium bifluoride (ABF) salt that enables the introduction of spikes as tracers or internal standards. Proof of principle experiments show that the decomposition of uranium ore in sealed PFA fluoropolymer vials at 230 °C yields, after cooling, new solids suitable for direct solid sampling by LA. When spikes are included in the molten salt reaction, subsequent LA-ICP-MS sampling at several spots indicate that the spikes are evenly distributed, and that U-235 tracer dramatically improves reproducibility in U-238 analysis. Precisions improved from 17% relative standard deviation for U-238 signals to 0.1% for the ratio of sample U-238 to spiked U-235, a factor of over two orders of magnitude. These results introduce the concept of solid matrix transformation (SMT) using ABF, and provide proof of principle for a new method of incorporating internal standards into a solid for LA-ICP-MS. This new approach, SMT-LA-ICP-MS, provides opportunities to improve calibration and quantification in solids based analysis. Looking forward, tracer addition to transformed solids opens up LA-based methods to analytical methodologies such as standard addition, isotope dilution, preparation of matrix-matched solid standards, external calibration, and monitoring instrument drift against external calibration standards.

  16. Improved sample preparation of glyphosate and methylphosphonic acid by EPA method 6800A and time-of-flight mass spectrometry using novel solid-phase extraction. (United States)

    Wagner, Rebecca; Wetzel, Stephanie J; Kern, John; Kingston, H M Skip


    The employment of chemical weapons by rogue states and/or terrorist organizations is an ongoing concern in the United States. The quantitative analysis of nerve agents must be rapid and reliable for use in the private and public sectors. Current methods describe a tedious and time-consuming derivatization for gas chromatography-mass spectrometry and liquid chromatography in tandem with mass spectrometry. Two solid-phase extraction (SPE) techniques for the analysis of glyphosate and methylphosphonic acid are described with the utilization of isotopically enriched analytes for quantitation via atmospheric pressure chemical ionization-quadrupole time-of-flight mass spectrometry (APCI-Q-TOF-MS) that does not require derivatization. Solid-phase extraction-isotope dilution mass spectrometry (SPE-IDMS) involves pre-equilibration of a naturally occurring sample with an isotopically enriched standard. The second extraction method, i-Spike, involves loading an isotopically enriched standard onto the SPE column before the naturally occurring sample. The sample and the spike are then co-eluted from the column enabling precise and accurate quantitation via IDMS. The SPE methods in conjunction with IDMS eliminate concerns of incomplete elution, matrix and sorbent effects, and MS drift. For accurate quantitation with IDMS, the isotopic contribution of all atoms in the target molecule must be statistically taken into account. This paper describes two newly developed sample preparation techniques for the analysis of nerve agent surrogates in drinking water as well as statistical probability analysis for proper molecular IDMS. The methods described in this paper demonstrate accurate molecular IDMS using APCI-Q-TOF-MS with limits of quantitation as low as 0.400 mg/kg for glyphosate and 0.031 mg/kg for methylphosphonic acid. Copyright © 2012 John Wiley & Sons, Ltd.

  17. Sampling system and method

    Energy Technology Data Exchange (ETDEWEB)

    Decker, David L.; Lyles, Brad F.; Purcell, Richard G.; Hershey, Ronald Lee


    In one embodiment, the present disclosure provides an apparatus and method for supporting a tubing bundle during installation or removal. The apparatus includes a clamp for securing the tubing bundle to an external wireline. In various examples, the clamp is external to the tubing bundle or integral with the tubing bundle. According to one method, a tubing bundle and wireline are deployed together and the tubing bundle periodically secured to the wireline using a clamp. In another embodiment, the present disclosure provides an apparatus and method for coupling conduit segments together. A first pump obtains a sample and transmits it through a first conduit to a reservoir accessible by a second pump. The second pump further conducts the sample from the reservoir through a second conduit. In a specific example, one or more clamps are used to connect the first and/or second conduits to an external wireline.

  18. Solubilization of proteins in extracted oil bodies by SDS: a simple and efficient protein sample preparation method for Tricine-SDS-PAGE. (United States)

    Ying, Yusang; Zhao, Luping; Kong, Lingzhi; Kong, Xiangzhen; Hua, Yufei; Chen, Yeming


    A simple and efficient method for preparing Tricine-SDS-PAGE protein sample of extracted oil bodies (OBs) was supplied: OB suspension was vortexed with SDS buffer (pH 6.8) for 2 min at room temperature with SDS/protein of 1.52/1(w/w), which could be analyzed by Tricine-SDS-PAGE after simple treatments (dilution and 2-mercaptoethanol). At SDS/protein of 1.52/1, about 95% of proteins in soybean OB suspension were solubilized, whereas residual 5% of proteins were weakly bound to SDS-destroyed OBs; proteins in destroyed OBs might be further solubilized by SDS in the gel and cathode buffer of Tricine-SDS-PAGE, causing about 99% of proteins in soybean OB suspension recover on Tricine-SDS-PAGE gel, which was better than acetone (89%) and diethyl ether (96%) harvested protein samples. Higher or lower SDS/protein was unbeneficial for protein solubilization from OBs. Additionally, the above method was also better than organic solvent method for peanut, sesame, and rapeseed OB suspensions. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Application of a Stir Bar Sorptive Extraction sample preparation method with HPLC for soil fungal biomass determination in soils from a detrital manipulation study. (United States)

    Beni, Áron; Lajtha, Kate; Kozma, János; Fekete, István


    Ergosterol is a sterol found ubiquitously in cell membranes of filamentous fungi. Although concentrations in different fungal species span the range of 2.6 to 42μg/mL of dry mass, many studies have shown a strong correlation between soil ergosterol content and fungal biomass. The analysis of ergosterol in soil therefore could be an effective tool for monitoring changes in fungal biomass under different environmental conditions. Stir Bar Sorptive Extraction (SBSE) is a new sample preparation method to extract and concentrate organic analytes from liquid samples. SBSE was here demonstrated to be a simple, fast, and cost effective method for the quantitative analysis of ergosterol from field-collected soils. Using this method we observed that soil ergosterol as a measure of fungal biomass proved to be a sensitive indicator of soil microbial dynamics that were altered by changes in plant detrital inputs to soils in a long-term field experiment. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Optimising uncertainty in physical sample preparation. (United States)

    Lyn, Jennifer A; Ramsey, Michael H; Damant, Andrew P; Wood, Roger


    Uncertainty associated with the result of a measurement can be dominated by the physical sample preparation stage of the measurement process. In view of this, the Optimised Uncertainty (OU) methodology has been further developed to allow the optimisation of the uncertainty from this source, in addition to that from the primary sampling and the subsequent chemical analysis. This new methodology for the optimisation of physical sample preparation uncertainty (u(prep), estimated as s(prep)) is applied for the first time, to a case study of myclobutanil in retail strawberries. An increase in expenditure (+7865%) on the preparatory process was advised in order to reduce the s(prep) by the 69% recommended. This reduction is desirable given the predicted overall saving, under optimised conditions, of 33,000 pounds Sterling per batch. This new methodology has been shown to provide guidance on the appropriate distribution of resources between the three principle stages of a measurement process, including physical sample preparation.

  1. Radioactive air sampling methods

    CERN Document Server

    Maiello, Mark L


    Although the field of radioactive air sampling has matured and evolved over decades, it has lacked a single resource that assimilates technical and background information on its many facets. Edited by experts and with contributions from top practitioners and researchers, Radioactive Air Sampling Methods provides authoritative guidance on measuring airborne radioactivity from industrial, research, and nuclear power operations, as well as naturally occuring radioactivity in the environment. Designed for industrial hygienists, air quality experts, and heath physicists, the book delves into the applied research advancing and transforming practice with improvements to measurement equipment, human dose modeling of inhaled radioactivity, and radiation safety regulations. To present a wide picture of the field, it covers the international and national standards that guide the quality of air sampling measurements and equipment. It discusses emergency response issues, including radioactive fallout and the assets used ...

  2. Materials of 2nd Poznan analytical meeting: Modern methods of sample preparation and trace elements analysis; Materialy 2. Poznanskiego Kowersatorium Analitycznego: Nowoczesne metody przygotowania probek i oznaczania sladowych zawartosci pierwiastkow

    Energy Technology Data Exchange (ETDEWEB)



    The modern methods for sample preparation and treatment before analysis have been presented. The neutron activation analysis, PIXE, X-ray fluorescence analysis and other methods have been used for trace element analysis in samples of different origin. The new technical solutions and modern equipment for trace amount analysis have been also performed. During the conference 20 lectures have been presented.

  3. 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


    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...

  4. Method validation and dissipation kinetics of four herbicides in maize and soil using QuEChERS sample preparation and liquid chromatography tandem mass spectrometry. (United States)

    Pang, Nannan; Wang, Tielong; Hu, Jiye


    A versatile liquid chromatography tandem mass spectrometry method with modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) sample preparation was developed for the determination of rimsulfuron, mesotrione, fluroxypyr-meptyl, and fluroxypyr. By adjusting the amount of graphitized carbon black, the herbicide analytes could be quantified with satisfactory recoveries in the range of 80-110%. A dissipation kinetics study conducted under open field conditions at two sites during 2014 showed first order equations with half-lives between 0.6d and 3.6d, illustrating an appropriate degree of stability and safety. The dissipation kinetics were different in the different matrices. Although the herbicides had higher initial residues in straw than those in soil, they degraded faster in straw. The terminal residues for the herbicides formulated in two water dispersible granules were all below maximum residue limits. These results not only gave insights about the analytes but also contributed to environmental protection and food safety. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. Preparation methods of alginate nanoparticles

    NARCIS (Netherlands)

    Paques, J.P.; Linden, van der E.; Rijn, van C.J.M.; Sagis, L.M.C.


    This article reviews available methods for the formation of alginate nano-aggregates, nanocapsules and nanospheres. Primarily, alginate nanoparticles are being prepared by two methods. In the “complexation method”, complex formation on the interface of an oil droplet is used to form alginate

  6. Development of a new sample preparation method based on liquid-liquid-liquid extraction combined with dispersive liquid-liquid microextraction and its application on unfiltered samples containing high content of solids. (United States)

    Farajzadeh, Mir Ali; Abbaspour, Maryam


    A new sample preparation method based on liquid-liquid-liquid extraction combined with dispersive liquid-liquid microextraction followed by gas chromatography-flame ionization detection has been reported for the extraction/preconcentration and determination of trace levels of twelve pesticide residues from different samples with high content of solids without filtration. This method consists of a three-phase system including an aqueous phase (sample solution), acetonitrile, and hexane. The extraction mechanism is based on different affinities of the substances from the sample matrices towards each of the involved phase, which provides a high selectivity to the process. In other words, interfering hydrophobic compounds are transferred into hexane and will not be present in the final extract. Furthermore, ionic and polar compounds are retained in the aqueous phase. Therefore, only semi-polar compounds such as the studied pesticides are extracted into acetonitrile. In this method, a homogeneous solution of the aqueous phase and acetonitrile (a water-soluble extraction solvent) forms two clearly separated phases in the presence of sodium sulfate (as a phase separation agent) and simultaneously the analytes are extracted into the fine droplets of the acetonitrile collected on the surface of the aqueous phase. To achieve high enrichment factors, the acetonitrile phase is mixed with 1,2-dibromoethane (as a preconcentration solvent) at µL-level to perform the following dispersive liquid-liquid microextraction procedure. Several parameters that can affect extraction efficiency including kind and volume of extraction solvent, type and concentration of phase separation agent, hexane volume, kind of preconcentration solvent, and ionic strength were studied and optimized. Under the optimal conditions, extraction recoveries were obtained in the range of 53-93% and the calibration curves were linear in wide ranges with correlation coefficients ≥ 0.9983. Intra- (n = 6) and

  7. Sample processing device and method

    DEFF Research Database (Denmark)


    A sample processing device is disclosed, which sample processing device comprises a first substrate and a second substrate, where the first substrate has a first surface comprising two area types, a first area type with a first contact angle with water and a second area type with a second contact...... a sample liquid comprising the sample and the first preparation system is adapted to receive a receiving liquid. In a particular embodiment, a magnetic sample transport component, such as a permanent magnet or an electromagnet, is arranged to move magnetic beads in between the first and second substrates....


    Katz, J.J.; Sheft, I.


    A method is presented for preparing the halides of elements which are relatively non-reactive with halogenating agents. The method involves reacting a mixture of an oxygen containing salt of a difficulty halogenated metal with an oxygen containing salt of an easily halogenated metal with a halogenating agent. Accordingly plutonium tetrafluoride is produced by reacting a mixture of plutonium dioxide and uranium octaoxide with bromine trifluoride. The reaction proceeds smoothly at moderate temperatures and the resulting plutonium trifluoride may be readily separated from many impurities which form volatile fluorides by volatilizing these volatile fluorides from the reaction chamber.

  9. Optimized preparation of urine samples for two-dimensional electrophoresis and initial application to patient samples

    DEFF Research Database (Denmark)

    Lafitte, Daniel; Dussol, Bertrand; Andersen, Søren


    OBJECTIVE: We optimized of the preparation of urinary samples to obtain a comprehensive map of urinary proteins of healthy subjects and then compared this map with the ones obtained with patient samples to show that the pattern was specific of their kidney disease. DESIGN AND METHODS: The urinary...

  10. [Evaluation of Two Methods (Nativ-Lugol Preparation and Enzyme-Linked Immunosorbent Assay) for Detection of Entamoeba histolytica in Stool Samples]. (United States)

    Alver, Oktay; Topaç, Tuncay; Töre, Okan


    This study aims to compare the performance of Native-Lugol examination and EIA Antigen Detection Test using stool samples obtained from patients diagnosed as clinical gastroenteritis and submitted to the Parasitology Laboratory in Uludağ University between January 2010 and February 2011. The stool samples taken from 116 patients and sent to the laboratory of parasitology from various clinics including outpatient services have been investigated using Native-Lugol examination and EIA Antigen Detection Kit (Wampole® E. histolytica II Techlab®, Inc., Blacksburg, Virginia) methods on all the samples. In one of 116 stool samples (%0,86), E. histolytica/E. dispar cysts and/or trophozoites were detected by using direct microscobic (nativ-lugol) method. E. histolytica specific antigen was detected in 34 (29.3%) out of the sample set, and the patients were given adequate treatment. The highest rate of E. histolytica specific antigen positivity were observed in 11-19 age group. On account of the fact that the sensitivity of direct microscopy is quite low, it is concluded that, from the viewpoint of preventing the amebiasis suspected patients from false diagnosis and hence from receiving inadequate treatment, the use of the ELISA method is more appropriate and advantageous, as it is cost effective and does not require highly qualified staff.

  11. 7 CFR 27.21 - Preparation of samples of cotton. (United States)


    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Preparation of samples of cotton. 27.21 Section 27.21... REGULATIONS COTTON CLASSIFICATION UNDER COTTON FUTURES LEGISLATION Regulations Inspection and Samples § 27.21 Preparation of samples of cotton. The samples from each bale shall be prepared as specified in this section...

  12. Rapid Automated Sample Preparation for Biological Assays

    Energy Technology Data Exchange (ETDEWEB)

    Shusteff, M


    Our technology utilizes acoustic, thermal, and electric fields to separate out contaminants such as debris or pollen from environmental samples, lyse open cells, and extract the DNA from the lysate. The objective of the project is to optimize the system described for a forensic sample, and demonstrate its performance for integration with downstream assay platforms (e.g. MIT-LL's ANDE). We intend to increase the quantity of DNA recovered from the sample beyond the current {approx}80% achieved using solid phase extraction methods. Task 1: Develop and test an acoustic filter for cell extraction. Task 2: Develop and test lysis chip. Task 3: Develop and test DNA extraction chip. All chips have been fabricated based on the designs laid out in last month's report.

  13. Determination of chemical oxygen demand in heterogeneous solid or semisolid samples using a novel method combining solid dilutions as a preparation step followed by optimized closed reflux and colorimetric measurement. (United States)

    Noguerol-Arias, Joan; Rodríguez-Abalde, Angela; Romero-Merino, Eva; Flotats, Xavier


    This paper reports the development of an innovative sample preparation method for the determination of the chemical oxygen demand (COD) in heterogeneous solid or semisolid samples, with high suspended solids and COD concentrations, using an optimized closed reflux colorimetric method. The novel method, named solid dilution (SD), is based on a different technique of sample preparation, diluting the sample with magnesium sulfate (MgSO(4)) previous to COD determination. With this, it is possible to obtain a solid homogeneous mixture much more easily analyzable. Besides, a modification of concentration and ratio of reagents was optimized to make the closed reflux colorimetric method suitable for complex substrates with COD levels ranging from 5 to 2500 g O(2) kg(-1) TS. The optimized method has been tested with potassium hydrogen phthalate (KHP) as primary solid standard and using different solid or semiliquid substrates like pig slaughterhouse waste and sewage sludge, among others. Finally, the optimized method (SD/SM-CRC) was intensively tested in comparison to the standard titrimetric method (SM-ORT) using different certified reference materials (CRM). The developed method was found to give higher accuracy, 1.4% relative standard deviation (RSD) vs 10.4%, and bias of 2.8% vs 8.0%, in comparison to the standard open reflux titrimetric method.

  14. Sample preparation for quantitation of tritium by accelerator mass spectrometry. (United States)

    Chiarappa-Zucca, Marina L; Dingley, Karen H; Roberts, Mark L; Velsko, Carol A; Love, Adam H


    The capability to prepare samples accurately and reproducibly for analysis of tritium (3H) content by accelerator mass spectrometry (AMS) greatly facilitates isotopic tracer studies in which attomole levels of 3H can be measured in milligram-sized samples. A method has been developed to convert the hydrogen of organic samples to a solid, titanium hydride, which can be analyzed by AMS. Using a two-step process, the sample is first oxidized to carbon dioxide and water. In the second step, the water is transferred within a heated manifold into a quartz tube, reduced to hydrogen gas using zinc, and reacted with titanium powder. The 3H/1H ratio of the titanium hydride is measured by AMS and normalized to standards whose ratios were determined by decay counting to calculate the amount of 3H in the original sample. Water, organic compounds, and biological samples with 3H activities measured by liquid scintillation counting were utilized to develop and validate the method. The 3H/1H ratios were quantified in samples that spanned 5 orders of magnitude, from 10(-10) to 10(-15), with a detection limit of 3.0 x 10(-15), which is equivalent to 0.02 dpm tritium/mg of material. Samples smaller than 2 mg were analyzed following addition of 2 mg of a tritium-free-hydrogen carrier. Preparation of organic standards containing both 14C and 3H in 2-mg organic samples demonstrated that this sample preparation methodology can also be applied to quantify both of these isotopes from a single sample.

  15. Congener Production in Blood Samples During Preparation and Storage

    DEFF Research Database (Denmark)

    Felby, Søren; Nielsen, Erik


    Retsmedicin, congener production, preparation, head space GC, acetone, isobutanol, storage, blood samples, n-propanol, methanol, methylethylketone......Retsmedicin, congener production, preparation, head space GC, acetone, isobutanol, storage, blood samples, n-propanol, methanol, methylethylketone...

  16. Enhanced sensitivity in PCR detection of Listeria monocytogenes in soft cheese through use of an aqueous two-phase system as a sample preparation method.


    Lantz, P G; Tjerneld, F; Borch, E; Hahn-Hägerdal, B; Rådström, P.


    A sample treatment method based on an aqueous two-phase system containing polyethylene glycol and dextran was developed for enhancing sensitivity in the detection of Listeria monocytogenes in soft cheese with PCR. The results suggest that the improved detection sensitivity following partitioning of the cheese homogenate in an aqueous two-phase system may be due to partitioning of the PCR inhibitors to the polyethylene glycol phase.

  17. Ammonia and ammonium over the southern Baltic Sea. Part 1. Preparation of aerosol and air samples for the determination of ammonia by the indophenol method

    Directory of Open Access Journals (Sweden)

    Lucyna Falkowska


    Full Text Available 1997 saw the start of a study into the spatial and temporal changes in the chemical composition of the atmosphere over the littoral and offshore waters of the Baltic Sea. Collection of samples and their subsequent chemical analysis was preceded by meticulous laboratory experiments concerning the validation and adaptation of the analytical procedures. The colorimetric indophenol blue technique was used to analyse the aerosol samples for the concentration of ammonium ions and the air samples for their gaseous ammonia. The samples were collected with PTFE-filters and Whatman 41 filters coated with 5% orthophosphoric acid. This acid enhances the aerosol sampling efficacy but reduces the pH of the reaction in which indophenol is formed. The pH of the aerosol samples thus had to be raised, and this in turn required an alteration to the original procedure prior to ammonia determination. It was demonstrated that the addition of 0.1 N KOH to the filters coated with H3PO4 increases the pH of the reaction medium to the required level of pH = 8-11.5 and does not substantially influence the precision of the determination; the error of the modified procedure was of the order of 5.2%.      Air samples for the determination of gaseous ammonia were collected with annular denuders. Oxalic acid and citric acid are the usual impregnating agents. In the present experiments oxalic acid was used for denuder impregnation; it turned out to be more effective than citric acid under the conditions of the southern Baltic Sea.      The detection limit of the indophenol blue method in these laboratory experiments was 0.045 mmol dm-3. The respective relative standard deviations (RSD within the range of higher and lower concentrations were 0.64% and 4.53%.

  18. Applications of Liquid-Phase Microextraction in the Sample Preparation of Environmental Solid Samples

    Directory of Open Access Journals (Sweden)

    Helena Prosen


    Full Text Available Solvent extraction remains one of the fundamental sample preparation techniques in the analysis of environmental solid samples, but organic solvents are toxic and environmentally harmful, therefore one of the possible greening directions is its miniaturization. The present review covers the relevant research from the field of application of microextraction to the sample preparation of environmental solid samples (soil, sediments, sewage sludge, dust etc. published in the last decade. Several innovative liquid-phase microextraction (LPME techniques that have emerged recently have also been applied as an aid in sample preparation of these samples: single-drop microextraction (SDME, hollow fiber-liquid phase microextraction (HF-LPME, dispersive liquid-liquid microextraction (DLLME. Besides the common organic solvents, surfactants and ionic liquids are also used. However, these techniques have to be combined with another technique to release the analytes from the solid sample into an aqueous solution. In the present review, the published methods were categorized into three groups: LPME in combination with a conventional solvent extraction; LPME in combination with an environmentally friendly extraction; LPME without previous extraction. The applicability of these approaches to the sample preparation for the determination of pollutants in solid environmental samples is discussed, with emphasis on their strengths, weak points and environmental impact.

  19. Neonatal blood gas sampling methods

    African Journals Online (AJOL)

    Blood gas sampling is part of everyday practice in the care of babies admitted to the neonatal intensive care unit, particularly for those receiving respiratory support. There is little published guidance that systematically evaluates the different methods of neonatal blood gas sampling, where each method has its individual ...

  20. Optimization for Peptide Sample Preparation for Urine Peptidomics

    Energy Technology Data Exchange (ETDEWEB)

    Sigdel, Tara K.; Nicora, Carrie D.; Hsieh, Szu-Chuan; Dai, Hong; Qian, Weijun; Camp, David G.; Sarwal, Minnie M.


    Analysis of native or endogenous peptides in biofluids can provide valuable insights into disease mechanisms. Furthermore, the detected peptides may also have utility as potential biomarkers for non-invasive monitoring of human diseases. The non-invasive nature of urine collection and the abundance of peptides in the urine makes analysis by high-throughput ‘peptidomics’ methods , an attractive approach for investigating the pathogenesis of renal disease. However, urine peptidomics methodologies can be problematic with regards to difficulties associated with sample preparation. The urine matrix can provide significant background interference in making the analytical measurements that it hampers both the identification of peptides and the depth of the peptidomics read when utilizing LC-MS based peptidome analysis. We report on a novel adaptation of the standard solid phase extraction (SPE) method to a modified SPE (mSPE) approach for improved peptide yield and analysis sensitivity with LC-MS based peptidomics in terms of time, cost, clogging of the LC-MS column, peptide yield, peptide quality, and number of peptides identified by each method. Expense and time requirements were comparable for both SPE and mSPE, but more interfering contaminants from the urine matrix were evident in the SPE preparations (e.g., clogging of the LC-MS columns, yellowish background coloration of prepared samples due to retained urobilin, lower peptide yields) when compared to the mSPE method. When we compared data from technical replicates of 4 runs, the mSPE method provided significantly improved efficiencies for the preparation of samples from urine (e.g., mSPE peptide identification 82% versus 18% with SPE; p = 8.92E-05). Additionally, peptide identifications, when applying the mSPE method, highlighted the biology of differential activation of urine peptidases during acute renal transplant rejection with distinct laddering of specific peptides, which was obscured for most proteins

  1. Culture conditions and sample preparation methods affect spectrum quality and reproducibility during profiling of Staphylococcus aureus with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. (United States)

    Goldstein, J E; Zhang, L; Borror, C M; Rago, J V; Sandrin, T R


    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has emerged as a promising tool to rapidly characterize Staphylococcus aureus. Different protocols have been employed, but effects of experimental factors, such as culture condition and sample preparation, on spectrum quality and reproducibility have not been rigorously examined. We applied MALDI-TOF MS to characterize a model system consisting of five methicillin-sensitive (MSSA) and five methicillin-resistant S. aureus isolates (MRSA) under two culture conditions (agar and broth) and using two sample preparation methods [intact cell method and protein extraction method (PEM)]. The effects of these treatments on spectrum quality and reproducibility were quantified. PEM facilitated increases in the number of peaks and mass range width. Broth cultures further improved spectrum quality in terms of increasing the number of peaks. In addition, PEM increased reproducibility in samples prepared using identical culture conditions. MALDI imaging data suggested that the improvement in reproducibility may result from a more homogeneous distribution of sample associated with the broth/PEM treatment. Broth/PEM treatment also yielded the highest rate (96%) of correct classification for MRSA. Taken together, these results suggest that broth/PEM maximizes the performance of MALDI-TOF MS to characterize S. aureus. Two culture conditions (agar or broth) and two sample preparation methods (intact cell or protein extraction) were evaluated for their effects on profiling of Staphylococcus aureus using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Results indicated that MALDI-enabled profiling of S. aureus is most effective when cultures are grown in broth and processed using a protein extraction-based approach. These findings should enhance future efforts to maximize the performance of this approach to characterize strains of S. aureus. © 2013

  2. Development and optimization of a novel sample preparation method cored on functionalized nanofibers mat-solid-phase extraction for the simultaneous efficient extraction of illegal anionic and cationic dyes in foods. (United States)

    Qi, Feifei; Jian, Ningge; Qian, Liangliang; Cao, Weixin; Xu, Qian; Li, Jian


    A simple and efficient three-step sample preparation method was developed and optimized for the simultaneous analysis of illegal anionic and cationic dyes (acid orange 7, metanil yellow, auramine-O, and chrysoidine) in food samples. A novel solid-phase extraction (SPE) procedure based on nanofibers mat (NFsM) was proposed after solvent extraction and freeze-salting out purification. The preferred SPE sorbent was selected from five functionalized NFsMs by orthogonal experimental design, and the optimization of SPE parameters was achieved through response surface methodology (RSM) based on the Box-Behnken design (BBD). Under the optimal conditions, the target analytes could be completely adsorbed by polypyrrole-functionalized polyacrylonitrile NFsM (PPy/PAN NFsM), and the eluent was directly analyzed by high-performance liquid chromatography-diode array detection (HPLC-DAD). The limits of detection (LODs) were between 0.002 and 0.01 mg kg-1, and satisfactory linearity with correlation coefficients (R > 0.99) for each dye in all samples was achieved. Compared with the Chinese standard method and the published methods, the proposed method was simplified greatly with much lower requirement of sorbent (5.0 mg) and organic solvent (2.8 mL) and higher sample preparation speed (10 min/sample), while higher recovery (83.6-116.5%) and precision (RSDs method, we have successfully detected illegal ionic dyes in three common representative foods: yellow croaker, soybean products, and chili seasonings. Graphical abstract Schematic representation of the process of the three-step sample preparation.

  3. Partial Discharge Characteristics of Polymer Nanocomposite Materials in Electrical Insulation: A Review of Sample Preparation Techniques, Analysis Methods, Potential Applications, and Future Trends (United States)

    Izzati, Wan Akmal; Adzis, Zuraimy; Shafanizam, Mohd


    Polymer nanocomposites have recently been attracting attention among researchers in electrical insulating applications from energy storage to power delivery. However, partial discharge has always been a predecessor to major faults and problems in this field. In addition, there is a lot more to explore, as neither the partial discharge characteristic in nanocomposites nor their electrical properties are clearly understood. By adding a small amount of weight percentage (wt%) of nanofillers, the physical, mechanical, and electrical properties of polymers can be greatly enhanced. For instance, nanofillers in nanocomposites such as silica (SiO2), alumina (Al2O3) and titania (TiO2) play a big role in providing a good approach to increasing the dielectric breakdown strength and partial discharge resistance of nanocomposites. Such polymer nanocomposites will be reviewed thoroughly in this paper, with the different experimental and analytical techniques used in previous studies. This paper also provides an academic review about partial discharge in polymer nanocomposites used as electrical insulating material from previous research, covering aspects of preparation, characteristics of the nanocomposite based on experimental works, application in power systems, methods and techniques of experiment and analysis, and future trends. PMID:24558326

  4. A review of sample preparation and its influence on pH determination in concrete samples

    Directory of Open Access Journals (Sweden)

    S. Manso


    Full Text Available If we are to monitor the chemical processes in cementitious materials, then pH assays in the pore solutions of cement pastes, mortars, and concretes are of key importance. However, there is no standard method that regulates the sample-preparation method for pH determination. The state-of-the-art of different methods for pH determination in cementitious materials is presented in this paper and the influence of sample preparation in each case. Moreover, an experimental campaign compares three different techniques for pH determination. Its results contribute to establishing a basic criterion to help researchers select the most suitable method, depending on the purpose of the research. A simple tool is described for selecting the easiest and the most economic pH determination method, depending on the objective; especially for researchers and those with limited experience in this field.


    National Research Council Canada - National Science Library



    .... Our article aims to study audit sampling in audit of financial statements. As an audit technique largely used, in both its statistical and nonstatistical form, the method is very important for auditors...

  6. C-STrap Sample Preparation Method--In-Situ Cysteinyl Peptide Capture for Bottom-Up Proteomics Analysis in the STrap Format.

    Directory of Open Access Journals (Sweden)

    Alexandre Zougman

    Full Text Available Recently we introduced the concept of Suspension Trapping (STrap for bottom-up proteomics sample processing that is based upon SDS-mediated protein extraction, swift detergent removal and rapid reactor-type protein digestion in a quartz depth filter trap. As the depth filter surface is made of silica, it is readily modifiable with various functional groups using the silane coupling chemistries. Thus, during the digest, peptides possessing specific features could be targeted for enrichment by the functionalized depth filter material while non-targeted peptides could be collected as an unbound distinct fraction after the digest. In the example presented here the quartz depth filter surface is functionalized with the pyridyldithiol group therefore enabling reversible in-situ capture of the cysteine-containing peptides generated during the STrap-based digest. The described C-STrap method retains all advantages of the original STrap methodology and provides robust foundation for the conception of the targeted in-situ peptide fractionation in the STrap format for bottom-up proteomics. The presented data support the method's use in qualitative and semi-quantitative proteomics experiments.

  7. Microfluidic Vortex Enhancement for on-Chip Sample Preparation

    Directory of Open Access Journals (Sweden)

    Anna Haller


    Full Text Available In the past decade a large amount of analysis techniques have been scaled down to the microfluidic level. However, in many cases the necessary sample preparation, such as separation, mixing and concentration, remains to be performed off-chip. This represents a major hurdle for the introduction of miniaturized sample-in/answer-out systems, preventing the exploitation of microfluidic’s potential for small, rapid and accurate diagnostic products. New flow engineering methods are required to address this hitherto insufficiently studied aspect. One microfluidic tool that can be used to miniaturize and integrate sample preparation procedures are microvortices. They have been successfully applied as microcentrifuges, mixers, particle separators, to name but a few. In this work, we utilize a novel corner structure at a sudden channel expansion of a microfluidic chip to enhance the formation of a microvortex. For a maximum area of the microvortex, both chip geometry and corner structure were optimized with a computational fluid dynamic (CFD model. Fluorescent particle trace measurements with the optimized design prove that the corner structure increases the size of the vortex. Furthermore, vortices are induced by the corner structure at low flow rates while no recirculation is observed without a corner structure. Finally, successful separation of plasma from human blood was accomplished, demonstrating a potential application for clinical sample preparation. The extracted plasma was characterized by a flow cytometer and compared to plasma obtained from a standard benchtop centrifuge and from chips without a corner structure.


    Directory of Open Access Journals (Sweden)



    Full Text Available Audit sampling involves the application of audit procedures to less than 100% of items within an account balance or class of transactions. Our article aims to study audit sampling in audit of financial statements. As an audit technique largely used, in both its statistical and nonstatistical form, the method is very important for auditors. It should be applied correctly for a fair view of financial statements, to satisfy the needs of all financial users. In order to be applied correctly the method must be understood by all its users and mainly by auditors. Otherwise the risk of not applying it correctly would cause loose of reputation and discredit, litigations and even prison. Since there is not a unitary practice and methodology for applying the technique, the risk of incorrectly applying it is pretty high. The SWOT analysis is a technique used that shows the advantages, disadvantages, threats and opportunities. We applied SWOT analysis in studying the sampling method, from the perspective of three players: the audit company, the audited entity and users of financial statements. The study shows that by applying the sampling method the audit company and the audited entity both save time, effort and money. The disadvantages of the method are difficulty in applying and understanding its insight. Being largely used as an audit method and being a factor of a correct audit opinion, the sampling method’s advantages, disadvantages, threats and opportunities must be understood by auditors.

  9. Distance sampling methods and applications

    CERN Document Server

    Buckland, S T; Marques, T A; Oedekoven, C S


    In this book, the authors cover the basic methods and advances within distance sampling that are most valuable to practitioners and in ecology more broadly. This is the fourth book dedicated to distance sampling. In the decade since the last book published, there have been a number of new developments. The intervening years have also shown which advances are of most use. This self-contained book covers topics from the previous publications, while also including recent developments in method, software and application. Distance sampling refers to a suite of methods, including line and point transect sampling, in which animal density or abundance is estimated from a sample of distances to detected individuals. The book illustrates these methods through case studies; data sets and computer code are supplied to readers through the book’s accompanying website.  Some of the case studies use the software Distance, while others use R code. The book is in three parts.  The first part addresses basic methods, the ...

  10. A Simple Preparation Method for Diphosphoimidazole

    DEFF Research Database (Denmark)

    Rosenberg, T.


    A simple method for the preparation of diphosphoimidazole is presented that involves direct phosphorylation of imidazole by phosphorus oxychloride in alkaline aqueous solution. Details are given on the use of diphosphoimidazole in preparing sodium phosphoramidate and certain phosphorylated amino...

  11. A novel method for preparing microplastic fibers (United States)

    Cole, Matthew


    Microscopic plastic (microplastic, 0.1 µm–5 mm) is a widespread pollutant impacting upon aquatic ecosystems across the globe. Environmental sampling has revealed synthetic fibers are prevalent in seawater, sediments and biota. However, microplastic fibers are rarely used in laboratory studies as they are unavailable for purchase and existing preparation techniques have limited application. To facilitate the incorporation of environmentally relevant microplastic fibers into future studies, new methods are required. Here, a novel cryotome protocol has been developed. Nylon, polyethylene terephthalate and polypropylene fibers (10–28 μm diameter) were aligned, embedded in water-soluble freezing agent, and sectioned (40–100 μm length) using a cryogenic microtome. Microplastic fibers were prepared to specified lengths (P microplastics, with widths similar to those observed in the natural environment, which could ultimately lead to a better understanding of the biological and ecological effects of microplastic debris in the environment. PMID:27694820

  12. A novel method for preparing microplastic fibers (United States)

    Cole, Matthew


    Microscopic plastic (microplastic, 0.1 µm-5 mm) is a widespread pollutant impacting upon aquatic ecosystems across the globe. Environmental sampling has revealed synthetic fibers are prevalent in seawater, sediments and biota. However, microplastic fibers are rarely used in laboratory studies as they are unavailable for purchase and existing preparation techniques have limited application. To facilitate the incorporation of environmentally relevant microplastic fibers into future studies, new methods are required. Here, a novel cryotome protocol has been developed. Nylon, polyethylene terephthalate and polypropylene fibers (10-28 μm diameter) were aligned, embedded in water-soluble freezing agent, and sectioned (40-100 μm length) using a cryogenic microtome. Microplastic fibers were prepared to specified lengths (P effects of microplastic debris in the environment.

  13. An improved extraction method of rapeseed oil sample preparation for the subsequent determination in it of azole class fungicides by gas chromatography

    Directory of Open Access Journals (Sweden)

    Mikhail F. Zayats


    Full Text Available The distribution of 19 azole class pesticides in hexane/aqueous–organic mixtures systems and rapeseed oil (or oil solution in hexane/organic solvents has been studied at 20 ± 1 °C. The distribution constants (P and coefficients (D between hydrocarbon and polar phase are calculated. It is found that all the studied pesticides are hydrophobic, i.e., in hexane–water system logP ≫ 0. Replacement of water by organic solvents results in sharp logP falling, and their values become negative. It is revealed that solutions of strong inorganic acids in anhydrous acetonitrile extract azole class pesticides from hexane and vegetable oils most fully and selectively. In particular, the acidification of acetonitrile causes a drop of D values in 50–2000 times for the majority of the studied pesticides. This phenomenon was used for the development of the improved technique for the quantitative analysis of a widely used azole class pesticides, which can be presented at trace levels in rapeseed oil. The proposed methodology is based on dissociation extraction (DE of azoles using perchloric acid in anhydrous acetonitrile, with following clean-up of acetonitrile extract from organic impurities by hexane and aqueous solution of dipotassium hydrogen orthophosphate, and final GC–ECD (gas chromatography with electron capture detection determination of azole fungicides. The values of obtained recoveries were between 85% and 115% with RSD values below 10%. The obtained limits of quantitation, ranged from 3.0 to 300 μg kg−1, are below the maximum residue levels (MRLs set by the European Union for the majority of pesticides. The developed method was successfully applied to different rapeseed oil samples.

  14. Sample preparation strategies for food and biological samples prior to nanoparticle detection and imaging

    DEFF Research Database (Denmark)

    Larsen, Erik Huusfeldt; Löschner, Katrin


    AFFF-ICP-MS fractograms, which corresponded to the enzymatic digests, showed a major nano-peak (about 80 % recovery of AgNPs spiked to the meat) plus new smaller peaks that eluted close to the void volume of the fractograms. Small, but significant shifts in retention time of AFFF peaks were observed......-ICP-MS analysis of their content of gold nanoparticles (AuNPs) was tested and compared with enzymatic sample preparation [3]. The results showed that the same results, with respect to the obtained number-based size distribution for AuNPs, were obtained for the two preparation methods. In contrast, the alkaline...

  15. Universal Sample Preparation Module for Molecular Analysis in Space Project (United States)

    National Aeronautics and Space Administration — Lynntech proposes to develop and demonstrate the ability of a compact, light-weight, and automated universal sample preparation module (USPM) to process samples from...

  16. Evaluation of Two Methods (Nativ-Lugol Preparation and Enzyme-Linked Immunosorbent Assay) for Detection of Entamoeba histolytica in Stool Samples

    National Research Council Canada - National Science Library

    Oktay Alver; Tuncay Topaç; Okan Töre


      Objective : This study aims to compare the performance of Native-Lugol examination and EIA Antigen Detection Test using stool samples obtained from patients diagnosed as clinical gastroenteritis and submitted...

  17. Detection of Salmonella enterica in meat in less than 5 hours by a low-cost and non-complex sample preparation method

    DEFF Research Database (Denmark)

    Fachmann, Mette Sofie Rousing; Löfström, Charlotta; Hoorfar, Jeffrey


    , and help contribute to improved food safety. While the cost of analysis and hands-on time of the presented rapid method were comparable to reference culture methods, the fast product release by this method can provide the meat industry with a competitive advantage. Not only will the abattoirs save costs...

  18. Sample preparation and EFTEM of Meat Samples for Nanoparticle Analysis in Food (United States)

    Lari, L.; Dudkiewicz, A.


    Nanoparticles are used in industry for personal care products and the preparation of food. In the latter application, their functions include the prevention of microbes' growth, increase of the foods nutritional value and sensory quality. EU regulations require a risk assessment of the nanoparticles used in foods and food contact materials before the products can reach the market. However, availability of validated analytical methodologies for detection and characterisation of the nanoparticles in food hampers appropriate risk assessment. As part of a research on the evaluation of the methods for screening and quantification of Ag nanoparticles in meat we have tested a new TEM sample preparation alternative to resin embedding and cryo-sectioning. Energy filtered TEM analysis was applied to evaluate thickness and the uniformity of thin meat layers acquired at increasing input of the sample demonstrating that the protocols used ensured good stability under the electron beam, reliable sample concentration and reproducibility.

  19. 7 CFR 61.34 - Drawing and preparation of sample. (United States)


    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Drawing and preparation of sample. 61.34 Section 61.34 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards... Cottonseed Samplers § 61.34 Drawing and preparation of sample. Each licensed cottonseed sampler shall draw...

  20. The Recent Developments in Sample Preparation for Mass Spectrometry-Based Metabolomics. (United States)

    Gong, Zhi-Gang; Hu, Jing; Wu, Xi; Xu, Yong-Jiang


    Metabolomics is a critical member in systems biology. Although great progress has been achieved in metabolomics, there are still some problems in sample preparation, data processing and data interpretation. In this review, we intend to explore the roles, challenges and trends in sample preparation for mass spectrometry- (MS-) based metabolomics. The newly emerged sample preparation methods were also critically examined, including laser microdissection, in vivo sampling, dried blood spot, microwave, ultrasound and enzyme-assisted extraction, as well as microextraction techniques. Finally, we provide some conclusions and perspectives for sample preparation in MS-based metabolomics.

  1. Electrokinetics for sample preparation of biological molecules in biological samples using microfluidic systems. (United States)

    Shallan, Aliaa I; Guijt, Rosanne M; Breadmore, Michael C


    Sample preparation is the first part of every analytical method, but is often considered only after the optimization of the method. It is traditionally performed using a range of techniques requiring extensive manual handling, with solid-phase extraction, liquid-liquid extraction, protein precipitation and ultracentrfiguation, among others, being used depending on the targets and the application. In this article, we will focus on alternatives based on electrokinetics for applications including sample clean-up, concentration and derivatization of large biological molecules (DNA, peptides and proteins) of diagnostic importance, as well as small molecules as a tool for therapeutic drug monitoring. This article describes these approaches in terms of mechanisms, applicability and potential to be integrated into a lab-on-a-chip device for directly processing biological samples. Examples dealing with treated or clean samples have been excluded except where they show exceptionally high value.

  2. Rapid sample preparation for detection and identification of avian influenza virus from chicken faecal samples using magnetic bead microsystem

    DEFF Research Database (Denmark)

    Dhumpa, Raghuram; Bu, Minqiang; Handberg, Kurt


    -PCR is a sensitive method for detection of AIV, it requires sample preparation including separation and purification of AIV and concentrate viral RNA. It is laborious and complex process especially for diagnosis using faecal sample. In this study, magnetic beads were used for immunoseparation of AIV in chicken...... faecal sample by a magnetic microsystem. Using this system, all the 16 hemagglutinin (H) and 9 neuraminidase (N) subtypes of AIV were separated and detected in spiked faecal samples using RT-PCR, without an RNA extraction step. This rapid sample preparation method can be integrated with a total analysis...

  3. The beauty of being (label-free: sample preparation methods for SWATH-MS and next-generation targeted proteomics [v2; ref status: indexed,

    Directory of Open Access Journals (Sweden)

    Jakob Vowinckel


    Full Text Available The combination of qualitative analysis with label-free quantification has greatly facilitated the throughput and flexibility of novel proteomic techniques. However, such methods rely heavily on robust and reproducible sample preparation procedures. Here, we benchmark a selection of in gel, on filter, and in solution digestion workflows for their application in label-free proteomics. Each procedure was associated with differing advantages and disadvantages. The in gel methods interrogated were cost effective, but were limited in throughput and digest efficiency. Filter-aided sample preparations facilitated reasonable processing times and yielded a balanced representation of membrane proteins, but led to a high signal variation in quantification experiments. Two in solution digest protocols, however, gave optimal performance for label-free proteomics. A protocol based on the detergent RapiGest led to the highest number of detected proteins at second-best signal stability, while a protocol based on acetonitrile-digestion, RapidACN, scored best in throughput and signal stability but came second in protein identification. In addition, we compared label-free data dependent (DDA and data independent (SWATH acquisition on a TripleTOF 5600 instrument. While largely similar in protein detection, SWATH outperformed DDA in quantification, reducing signal variation and markedly increasing the number of precisely quantified peptides.

  4. The beauty of being (label-free: sample preparation methods for SWATH-MS and next-generation targeted proteomics [v1; ref status: indexed,

    Directory of Open Access Journals (Sweden)

    Jakob Vowinckel


    Full Text Available The combination of qualitative analysis with label-free quantification has greatly facilitated the throughput and flexibility of novel proteomic techniques. However, such methods rely heavily on robust and reproducible sample preparation procedures. Here, we benchmark a selection of in gel, on filter, and in solution digestion workflows for their application in label-free proteomics. Each procedure was associated with differing advantages and disadvantages. The in gel methods interrogated were cost effective, but were limited in throughput and digest efficiency. Filter-aided sample preparations facilitated reasonable processing times and yielded a balanced representation of membrane proteins, but led to a high signal variation in quantification experiments. Two in solution digest protocols, however, gave optimal performance for label-free proteomics. A protocol based on the detergent RapiGest led to the highest number of detected proteins at second-best signal stability, while a protocol based on acetonitrile-digestion, RapidACN, scored best in throughput and signal stability but came second in protein identification. In addition, we compared label-free data dependent (DDA and data independent (SWATH acquisition. While largely similar in protein detection, SWATH outperformed DDA in quantification, reducing signal variation and markedly increasing the number of precisely quantified peptides.

  5. Preparation and method of study of fossil diatoms

    Digital Repository Service at National Institute of Oceanography (India)

    Setty, M.G.A.P.

    to be exercised during the preparation of the samples A method for the preparation of strewn slides is then explained A detailed procedure for the study of diatom-strewn slides under the microscope, using the "England finder", is described, and a note on diatom...

  6. A review of analytical methods for the determination of four new phosphodiesterase type 5 inhibitors in biological samples and pharmaceutical preparations

    Directory of Open Access Journals (Sweden)

    Cristiane Franco Codevilla


    Full Text Available The introduction of oral phosphodiesterase type 5 inhibitor therapy in 1998 revolutionized the treatment of erectile dysfunction. Erectile dysfunction is the most common sexual problem in men. It often has a profound effect on intimate relationships and quality of life. The analysis of pharmaceuticals is an important part of the drug development process as well as for routine analysis and quality control of commercial formulations. Whereas the determination of sildenafil citrate, vardenafil and tadalafil are well documented by a variety of methods, there are few publications about the determination of udenafil, lodenafil carbonate, mirodenafil and avanafil. The paper presents a brief review of the action mechanism, adverse effects, pharmacokinetics and the most recent analytical methods that can determine drug concentration in biological matrices and pharmaceutical formulations of these four drugs.A introdução da terapia oral com inibidores da fosfodiesterase tipo 5, em 1998, revolucionou o tratamento da disfunção erétil. A disfunção erétil é o problema sexual mais comum em homens. Muitas vezes tem um efeito profundo nas relações íntimas e na qualidade de vida. A análise de produtos farmacêuticos é uma parte importante do processo de desenvolvimento de fármacos, bem como para a análise de rotina e controle de qualidade das formulações comerciais. Enquanto a determinação do citrato de sildenafila, vardenafila e tadalafila está bem documentada por uma variedade de métodos, existem poucas publicações sobre a determinação de udenafila, carbonato de lodenafila, mirodenafila e avanafila. O artigo apresenta uma breve revisão do mecanismo de ação, efeitos adversos, farmacocinética e os mais recentes métodos analíticos, que podem determinar a concentração do fármaco em matrizes biológicas e formulações farmacêuticas destes quatro fármacos.

  7. Preparation of Cytology Samples: Tricks of the Trade. (United States)

    Moore, A Russell


    General principles and techniques for collection, preparation, and staining of cytologic samples in the general practice setting are reviewed. Tips for collection of digital images are also discussed. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. FISHprep: A Novel Integrated Device for Metaphase FISH Sample Preparation

    DEFF Research Database (Denmark)

    Shah, Pranjul Jaykumar; Vedarethinam, Indumathi; Kwasny, Dorota


    We present a novel integrated device for preparing metaphase chromosomes spread slides (FISHprep). The quality of cytogenetic analysis from patient samples greatly relies on the efficiency of sample pre-treatment and/or slide preparation. In cytogenetic slide preparation, cell cultures...... are routinely used to process samples (for culture, arrest and fixation of cells) and/or to expand limited amount of samples (in case of prenatal diagnostics). Arguably, this expansion and other sample pretreatments form the longest part of the entire diagnostic protocols spanning over 3–4 days. We present here...... a novel device with an integrated expansion chamber to culture, arrest and fix metaphase cells followed by a subsequent splashing protocol leading to ample metaphase chromosome spreads on a glass slide for metaphase FISH analysis. The device provides an easy, disposable, low cost, integrated solution...

  9. Sample Preparation for Electron Probe Microanalysis-Pushing the Limits. (United States)

    Geller, Joseph D; Engle, Paul D


    There are two fundamental considerations in preparing samples for electron probe microanalysis (EPMA). The first one may seem obvious, but we often find it is overlooked. That is, the sample analyzed should be representative of the population from which it comes. The second is a direct result of the assumptions in the calculations used to convert x-ray intensity ratios, between the sample and standard, to concentrations. Samples originate from a wide range of sources. During their journey to being excited under the electron beam for the production of x rays there are many possibilities for sample alteration. Handling can contaminate samples by adding extraneous matter. In preparation, the various abrasives used in sizing the sample by sawing, grinding and polishing can embed themselves. The most accurate composition of a contaminated sample is, at best, not representative of the original sample; it is misleading. Our laboratory performs EPMA analysis on customer submitted samples and prepares over 250 different calibration standards including pure elements, compounds, alloys, glasses and minerals. This large variety of samples does not lend itself to mass production techniques, including automatic polishing. Our manual preparation techniques are designed individually for each sample. The use of automated preparation equipment does not lend itself to this environment, and is not included in this manuscript. The final step in quantitative electron probe microanalysis is the conversion of x-ray intensities ratios, known as the "k-ratios," to composition (in mass fraction or atomic percent) and/or film thickness. Of the many assumptions made in the ZAF (where these letters stand for atomic number, absorption and fluorescence) corrections the localized geometry between the sample and electron beam, or takeoff angle, must be accurately known. Small angular errors can lead to significant errors in the final results. The sample preparation technique then becomes very

  10. Sample Preparation for Electron Probe Microanalysis—Pushing the Limits (United States)

    Geller, Joseph D.; Engle, Paul D.


    There are two fundamental considerations in preparing samples for electron probe microanalysis (EPMA). The first one may seem obvious, but we often find it is overlooked. That is, the sample analyzed should be representative of the population from which it comes. The second is a direct result of the assumptions in the calculations used to convert x-ray intensity ratios, between the sample and standard, to concentrations. Samples originate from a wide range of sources. During their journey to being excited under the electron beam for the production of x rays there are many possibilities for sample alteration. Handling can contaminate samples by adding extraneous matter. In preparation, the various abrasives used in sizing the sample by sawing, grinding and polishing can embed themselves. The most accurate composition of a contaminated sample is, at best, not representative of the original sample; it is misleading. Our laboratory performs EPMA analysis on customer submitted samples and prepares over 250 different calibration standards including pure elements, compounds, alloys, glasses and minerals. This large variety of samples does not lend itself to mass production techniques, including automatic polishing. Our manual preparation techniques are designed individually for each sample. The use of automated preparation equipment does not lend itself to this environment, and is not included in this manuscript. The final step in quantitative electron probe microanalysis is the conversion of x-ray intensities ratios, known as the “k-ratios,” to composition (in mass fraction or atomic percent) and/or film thickness. Of the many assumptions made in the ZAF (where these letters stand for atomic number, absorption and fluorescence) corrections the localized geometry between the sample and electron beam, or takeoff angle, must be accurately known. Small angular errors can lead to significant errors in the final results. The sample preparation technique then becomes very

  11. Novel Sample-handling Approach for XRD Analysis with Minimal Sample Preparation (United States)

    Sarrazin, P.; Chipera, S.; Bish, D.; Blake, D.; Feldman, S.; Vaniman, D.; Bryson, C.


    Sample preparation and sample handling are among the most critical operations associated with X-ray diffraction (XRD) analysis. These operations require attention in a laboratory environment, but they become a major constraint in the deployment of XRD instruments for robotic planetary exploration. We are developing a novel sample handling system that dramatically relaxes the constraints on sample preparation by allowing characterization of coarse-grained material that would normally be impossible to analyze with conventional powder-XRD techniques.

  12. Selecting Sample Preparation Workflows for Mass Spectrometry-Based Proteomic and Phosphoproteomic Analysis of Patient Samples with Acute Myeloid Leukemia

    Directory of Open Access Journals (Sweden)

    Maria Hernandez-Valladares


    Full Text Available Global mass spectrometry (MS-based proteomic and phosphoproteomic studies of acute myeloid leukemia (AML biomarkers represent a powerful strategy to identify and confirm proteins and their phosphorylated modifications that could be applied in diagnosis and prognosis, as a support for individual treatment regimens and selection of patients for bone marrow transplant. MS-based studies require optimal and reproducible workflows that allow a satisfactory coverage of the proteome and its modifications. Preparation of samples for global MS analysis is a crucial step and it usually requires method testing, tuning and optimization. Different proteomic workflows that have been used to prepare AML patient samples for global MS analysis usually include a standard protein in-solution digestion procedure with a urea-based lysis buffer. The enrichment of phosphopeptides from AML patient samples has previously been carried out either with immobilized metal affinity chromatography (IMAC or metal oxide affinity chromatography (MOAC. We have recently tested several methods of sample preparation for MS analysis of the AML proteome and phosphoproteome and introduced filter-aided sample preparation (FASP as a superior methodology for the sensitive and reproducible generation of peptides from patient samples. FASP-prepared peptides can be further fractionated or IMAC-enriched for proteome or phosphoproteome analyses. Herein, we will review both in-solution and FASP-based sample preparation workflows and encourage the use of the latter for the highest protein and phosphorylation coverage and reproducibility.

  13. Sampling of illicit drugs for quantitative analysis--part III: sampling plans and sample preparations. (United States)

    Csesztregi, T; Bovens, M; Dujourdy, L; Franc, A; Nagy, J


    The findings in this paper are based on the results of our drug homogeneity studies and particle size investigations. Using that information, a general sampling plan (depicted in the form of a flow-chart) was devised that could be applied to the quantitative instrumental analysis of the most common illicit drugs: namely heroin, cocaine, amphetamine, cannabis resin, MDMA tablets and herbal cannabis in 'bud' form (type I). Other more heterogeneous forms of cannabis (type II) were found to require alternative, more traditional sampling methods. A table was constructed which shows the sampling uncertainty expected when a particular number of random increments are taken and combined to form a single primary sample. It also includes a recommended increment size; which is 1 g for powdered drugs and cannabis resin, 1 tablet for MDMA and 1 bud for herbal cannabis in bud form (type I). By referring to that table, individual laboratories can ensure that the sampling uncertainty for a particular drug seizure can be minimised, such that it lies in the same region as their analytical uncertainty for that drug. The table shows that assuming a laboratory wishes to quantitatively analyse a seizure of powdered drug or cannabis resin with a 'typical' heterogeneity, a primary sample of 15×1 g increments is generally appropriate. The appropriate primary sample for MDMA tablets is 20 tablets, while for herbal cannabis (in bud form) 50 buds were found to be appropriate. Our study also showed that, for a suitably homogenised primary sample of the most common powdered drugs, an analytical sample size of between 20 and 35 mg was appropriate and for herbal cannabis the appropriate amount was 200 mg. The need to ensure that the results from duplicate or multiple incremental sampling were compared, to demonstrate whether or not a particular seized material has a 'typical' heterogeneity and that the sampling procedure applied has resulted in a 'correct sample', was highlighted and the setting

  14. Sample Preparation of Corn Seed Tissue to Prevent Analyte Relocations for Mass Spectrometry Imaging (United States)

    Kim, Shin Hye; Kim, Jeongkwon; Lee, Young Jin; Lee, Tae Geol; Yoon, Sohee


    Corn seed tissue sections were prepared by the tape support method using an adhesive tape, and mass spectrometry imaging (MSI) was performed. The effect of heat generated during sample preparation was investigated by time-of-flight secondary mass spectrometry (TOF-SIMS) imaging of corn seed tissue prepared by the tape support and the thaw-mounted methods. Unlike thaw-mounted sample preparation, the tape support method does not cause imaging distortion because of the absence of heat, which can cause migration of the analytes on the sample. By applying the tape-support method, the corn seed tissue was prepared without structural damage and MSI with accurate spatial information of analytes was successfully performed.

  15. Towards Cost-efficient Sampling Methods

    CERN Document Server

    Peng, Luo; Chong, Wu


    The sampling method has been paid much attention in the field of complex network in general and statistical physics in particular. This paper presents two new sampling methods based on the perspective that a small part of vertices with high node degree can possess the most structure information of a network. The two proposed sampling methods are efficient in sampling the nodes with high degree. The first new sampling method is improved on the basis of the stratified random sampling method and selects the high degree nodes with higher probability by classifying the nodes according to their degree distribution. The second sampling method improves the existing snowball sampling method so that it enables to sample the targeted nodes selectively in every sampling step. Besides, the two proposed sampling methods not only sample the nodes but also pick the edges directly connected to these nodes. In order to demonstrate the two methods' availability and accuracy, we compare them with the existing sampling methods in...

  16. Sample Preparation (SS): SE51_SS01 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available e Master NEO, BMS, Tokyo, Japan), and the seed powder was extracted with 1 mL of extraction buffer (0.1% HCO...trifugation (4 ℃, 10,000 rpm, 5 min), the sample tubes were subjected to sample preparation (buffer transfer

  17. Optimal sampling and sample preparation for NIR-based prediction of field scale soil properties (United States)

    Knadel, Maria; Peng, Yi; Schelde, Kirsten; Thomsen, Anton; Deng, Fan; Humlekrog Greve, Mogens


    The representation of local soil variability with acceptable accuracy and precision is dependent on the spatial sampling strategy and can vary with a soil property. Therefore, soil mapping can be expensive when conventional soil analyses are involved. Visible near infrared spectroscopy (vis-NIR) is considered a cost-effective method due to labour savings and relative accuracy. However, savings may be offset by the costs associated with number of samples and sample preparation. The objective of this study was to find the most optimal way to predict field scale total organic carbon (TOC) and texture. To optimize the vis-NIR calibrations the effects of sample preparation and number of samples on the predictive ability of models with regard to the spatial distribution of TOC and texture were investigated. Conditioned Latin hypercube sampling (cLHs) method was used to select 125 sampling locations from an agricultural field in Denmark, using electromagnetic induction (EMI) and digital elevation model (DEM) data. The soil samples were scanned in three states (field moist, air dried and sieved to 2 mm) with a vis-NIR spectrophotometer (LabSpec 5100, ASD Inc., USA). The Kennard-Stone algorithm was applied to select 50 representative soil spectra for the laboratory analysis of TOC and texture. In order to investigate how to minimize the costs of reference analysis, additional smaller subsets (15, 30 and 40) of samples were selected for calibration. The performance of field calibrations using spectra of soils at the three states as well as using different numbers of calibration samples was compared. Final models were then used to predict the remaining 75 samples. Maps of predicted soil properties where generated with Empirical Bayesian Kriging. The results demonstrated that regardless the state of the scanned soil, the regression models and the final prediction maps were similar for most of the soil properties. Nevertheless, as expected, models based on spectra from field

  18. Development and evaluation of microwave-assisted and ultrasound-assisted methods based on a quick, easy, cheap, effective, rugged, and safe sample preparation approach for the determination of bisphenol analogues in serum and sediments. (United States)

    Song, Shanjun; Shao, Mingwu; Wang, Weihua; He, Yajuan; Dai, Xinhua; Wang, Huiyu; Liu, Liliang; Guo, Feng


    Microwave- and ultrasound-assisted methods based on a quick, easy, cheap, effective, rugged, and safe sample preparation approach followed by high-performance liquid chromatography with tandem mass spectrometry were developed for the simultaneous determination of eight bisphenol analogues in serum and sediment. The developed methods provided satisfactory extraction efficiency for the energy provided by microwaves and ultrasound. Compositions of commercial sorbents (primary secondary amine, MgSO4 , octadecyl-modified silica, and graphitized carbon black) were evaluated. The ultrasound-assisted method was suited for serum using primary secondary amine, MgSO4 , and octadecyl-modified silica as sorbents and a mixture of hexane and ethyl acetate as extraction solvent. The microwave-assisted method worked better for sediment with tetrahydrofuran and methanol as solvents and primary secondary amine, MgSO4 , octadecyl-modified silica, and graphitized carbon black as sorbents. Other experimental parameters, such as extraction temperature and time, were also optimized. The inter- and intraday relative standard deviations ranged from 2.7 to 5.5%. The limits of detection were between 0.1 and 1.0 ng/mL for serum and between 0.1 and 0.5 ng/g dry weight for sediment. The proposed methods were successfully applied to seven sediment and 20 human serum samples. The results showed that the developed methods were practical for the analysis and biomonitoring of bisphenols in sera and sediment. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Optimised sample preparation of synovial fluid for detection of Chlamydia trachomatis DNA by polymerase chain reaction (United States)

    Kuipers, J.; Nietfeld, L.; Dreses-Werringloe..., U.; Koehler, L.; Wollenhaupt, J.; Zeidler, H.; Hammer, M.


    OBJECTIVE—To optimise sample preparation of synovial fluid for Chlamydia trachomatis (CT) specific polymerase chain reaction (PCR).
METHODS—Serial dilutions of purified CT elementary bodies in synovial fluid were prepared. The synovial fluid pellet was processed by eight different methods of sample preparation. Then samples were analysed by CT specific PCR. The sensitivity of PCR was the basis of ranking of the eight different methods.
RESULTS—Highest sensitivity was achieved by methods including an additional step of DNA isolation. Additional extraction of protein and polysaccharides by cetyltrimethylammonium bromide (CTAB) increased sensitivity. Addition of hyaluronidase did not increase sensitivity of QIAEX-DNA extraction but was necessary, however, before phenol-chloroform-DNA extraction.
CONCLUSIONS—The method of synovial fluid sample preparation significantly influences the sensitivity of subsequent PCR. Additional DNA isolation and extraction of PCR inhibitors by CTAB led to higher sensitivity.

 Keywords: Chlamydia trachomatis; polymerase chain reaction; synovial fluid PMID:10343525

  20. Identification of chemicals relevant to the Chemical Weapons Convention using the novel sample-preparation methods and strategies of the Mobile Laboratory of the Organization for the Prohibition of Chemical Weapons

    NARCIS (Netherlands)

    Terzic, O.; Gregg, H.; de Voogt, P.


    The standard approach to on-site sample preparation for gas chromatography-mass spectrometry analysis of chemicals relevant to the Chemical Weapons Convention provides relatively good coverage of the target analytes, but it suffers from a number of drawbacks, such as low sample throughput, use of

  1. Advanced Curation Preparation for Mars Sample Return and Cold Curation (United States)

    Fries, M. D.; Harrington, A. D.; McCubbin, F. M.; Mitchell, J.; Regberg, A. B.; Snead, C.


    NASA Curation is tasked with the care and distribution of NASA's sample collections, such as the Apollo lunar samples and cometary material collected by the Stardust spacecraft. Curation is also mandated to perform Advanced Curation research and development, which includes improving the curation of existing collections as well as preparing for future sample return missions. Advanced Curation has identified a suite of technologies and techniques that will require attention ahead of Mars sample return (MSR) and missions with cold curation (CCur) requirements, perhaps including comet sample return missions.

  2. An improved calcium chloride method preparation and ...

    African Journals Online (AJOL)

    In this paper, we have reported a modified method for preparation and transformation of competent cells. This modified method, improved from a classical protocol, has made some modifications on the concentration of calcium chloride and competent bacteria solution, rotation speed in centrifugation and centrifugation time.

  3. Low platinum catalyst and method of preparation

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Di-Jia; Chong, Lina


    A low platinum catalyst and method for making same. The catalyst comprises platinum-transition metal bimetallic alloy microcrystallites over a transition metal-nitrogen-carbon composite. A method of making a catalyst comprises preparation of transition metal organic frameworks, infusion of platinum, thermal treatment, and reduction to form the microcrystallites and composite.

  4. An improved calcium chloride method preparation and ...

    African Journals Online (AJOL)



    Dec 13, 2010 ... Transformation is one of the fundamental and essential molecular cloning techniques. In this paper, we have reported a modified method for preparation and transformation of competent cells. This modified method, improved from a classical protocol, has made some modifications on the concentration of.

  5. Sample preparation with solid phase microextraction and exhaustive extraction approaches: Comparison for challenging cases. (United States)

    Boyacı, Ezel; Rodríguez-Lafuente, Ángel; Gorynski, Krzysztof; Mirnaghi, Fatemeh; Souza-Silva, Érica A; Hein, Dietmar; Pawliszyn, Janusz


    In chemical analysis, sample preparation is frequently considered the bottleneck of the entire analytical method. The success of the final method strongly depends on understanding the entire process of analysis of a particular type of analyte in a sample, namely: the physicochemical properties of the analytes (solubility, volatility, polarity etc.), the environmental conditions, and the matrix components of the sample. Various sample preparation strategies have been developed based on exhaustive or non-exhaustive extraction of analytes from matrices. Undoubtedly, amongst all sample preparation approaches, liquid extraction, including liquid-liquid (LLE) and solid phase extraction (SPE), are the most well-known, widely used, and commonly accepted methods by many international organizations and accredited laboratories. Both methods are well documented and there are many well defined procedures, which make them, at first sight, the methods of choice. However, many challenging tasks, such as complex matrix applications, on-site and in vivo applications, and determination of matrix-bound and free concentrations of analytes, are not easily attainable with these classical approaches for sample preparation. In the last two decades, the introduction of solid phase microextraction (SPME) has brought significant progress in the sample preparation area by facilitating on-site and in vivo applications, time weighted average (TWA) and instantaneous concentration determinations. Recently introduced matrix compatible coatings for SPME facilitate direct extraction from complex matrices and fill the gap in direct sampling from challenging matrices. Following introduction of SPME, numerous other microextraction approaches evolved to address limitations of the above mentioned techniques. There is not a single method that can be considered as a universal solution for sample preparation. This review aims to show the main advantages and limitations of the above mentioned sample

  6. Sample preparation for mass spectrometry imaging of plant tissues: a review

    Directory of Open Access Journals (Sweden)

    Yonghui eDong


    Full Text Available Mass spectrometry imaging (MSI is a mass spectrometry based molecular ion imaging technique. It provides the means for ascertaining the spatial distribution of a large variety of analytes directly on tissue sample surfaces without any labeling or staining steps. These advantages make it an attractive molecular histology tool in medical, pharmaceutical and biological research. Likewise, MSI has started gaining popularity in plant sciences; yet, information regarding sample preparation methods specific for plant tissues is still limited. Sample preparation is a crucial step that is directly associated with the quality and authenticity of the imaging results, it therefore demands in-depth studies based on the characteristics of plant samples. In this review, a sample preparation pipeline is discussed in detail and illustrated through selected practical examples. In particular, special concerns regarding sample preparation for plant imaging are critically evaluated. Finally, the applications of various MSI techniques in plants are reviewed according to different classes of plant metabolites.

  7. Sample preparation for thermo-gravimetric determination and thermo-gravimetric characterization of refuse derived fuel. (United States)

    Robinson, T; Bronson, B; Gogolek, P; Mehrani, P


    Thermo-gravimetric analysis (TGA) is a useful method for characterizing fuels. In the past it has been applied to the study of refuse derived fuel (RDF) and related materials. However, the heterogeneity of RDF makes the preparation of small representative samples very difficult and this difficulty has limited the effectiveness of TGA for characterization of RDF. A TGA method was applied to a variety of materials prepared from a commercially available RDF using a variety of procedures. Applicability of TGA method to the determination of the renewable content of RDF was considered. Cryogenic ball milling was found to be an effective means of preparing RDF samples for TGA. When combined with an effective sample preparation, TGA could be used as an alternative method for assessing the renewable content of RDF. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

  8. Sample preparation issues in NMR-based plant metabolomics: optimisation for Vitis wood samples. (United States)

    Halabalaki, Maria; Bertrand, Samuel; Stefanou, Anna; Gindro, Katia; Kostidis, Sarantos; Mikros, Emmanuel; Skaltsounis, Leandros A; Wolfender, Jean-Luc


    Nuclear magnetic resonance (NMR) is one of the most commonly used analytical techniques in plant metabolomics. Although this technique is very reproducible and simple to implement, sample preparation procedures have a great impact on the quality of the metabolomics data. Investigation of different sample preparation methods and establishment of an optimised protocol for untargeted NMR-based metabolomics of Vitis vinifera L. wood samples. Wood samples from two different cultivars of V. vinifera with well-defined phenotypes (Gamaret and 2091) were selected as reference materials. Different extraction solvents (successively, dichloromethane, methanol and water, as well as ethyl acetate and 7:3 methanol-water (v/v)) and deuterated solvents (methanol-d4, 7:3 chloroform-d-methanol-d4 (v/v), dimethylsulphoxide-d6 and 9:1 dimethylsulphoxide-d6-water-d2 (v/v)) were evaluated for NMR acquisition, and the spectral quality was compared. The optimal extract concentration, chemical shift stability and peak area repeatability were also investigated. Ethyl acetate was found to be the most satisfactory solvent for the extraction of all representative chemical classes of secondary metabolites in V. vinifera wood. The optimal concentration of dried extract was 10 mg/mL and 7:3 chloroform-d-methanol-d4 (v/v) was the most suitable solvent system for NMR analysis. Multivariate data analysis was used to estimate the biological variation and clustering between different cultivars. Close attention should be paid to all required procedures before NMR analysis, especially to the selection of an extraction solvent and a deuterated solvent system to perform an extensive metabolomic survey of the specific matrix. Copyright © 2014 John Wiley & Sons, Ltd.

  9. Dissipation behavior of phorate and its toxic metabolites in the sandy clay loam soil of a tropical sugarcane ecosystem using a single-step sample preparation method and GC-MS. (United States)

    Ramasubramanian, Thirumalaiandi; Paramasivam, Mariappan


    The dissipation of phorate in the sandy clay loam soil of tropical sugarcane ecosystem was studied by employing a single-step sample preparation method and gas chromatography with mass spectrometry. The limit of quantification of the method was 0.01 μg/g. The recoveries of phorate, phorate sulfoxide, phorate sulfone, and phorate oxon were in the range 94.00-98.46% with relative standard deviations of 1.51-3.56% at three levels of fortification between 0.01 and 0.1 μg/g. The Half-life of phorate and the total residues, which include phorate, phorate sulfoxide and phorate sulfone, was 5.5 and 19.8 days, respectively at the recommended dose of insecticide. Phorate rapidly oxidized into its sulfoxide metabolite in the sandy clay loam soil. Phorate sulfoxide alone accounted for more than 20% of the total residues within 2 h post-application and it was more than 50% on the fifth day after treatment irrespective of the doses applied. Phorate sulfoxide and phorate sulfone reached below the detectable level on 105 and 135 days after treatment, respectively as against 45 days after treatment for phorate residues at the recommended dose. Thus, the reasonably prolonged efficacy of phorate against soil pests may be attributed to longer persistence of its more toxic sulfoxide and sulfone metabolites. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Sample Preparation Report of the Fourth OPCW Confidence Building Exercise on Biomedical Sample Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Udey, R. N. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Corzett, T. H. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Alcaraz, A. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)


    Following the successful completion of the 3rd biomedical confidence building exercise (February 2013 – March 2013), which included the analysis of plasma and urine samples spiked at low ppb levels as part of the exercise scenario, another confidence building exercise was targeted to be conducted in 2014. In this 4th exercise, it was desired to focus specifically on the analysis of plasma samples. The scenario was designed as an investigation of an alleged use of chemical weapons where plasma samples were collected, as plasma has been reported to contain CWA adducts which remain present in the human body for several weeks (Solano et al. 2008). In the 3rd exercise most participants used the fluoride regeneration method to analyze for the presence of nerve agents in plasma samples. For the 4th biomedical exercise it was decided to evaluate the analysis of human plasma samples for the presence/absence of the VX adducts and aged adducts to blood proteins (e.g., VX-butyrylcholinesterase (BuChE) and aged BuChE adducts using a pepsin digest technique to yield nonapeptides; or equivalent). As the aging of VX-BuChE adducts is relatively slow (t1/2 = 77 hr at 37 °C [Aurbek et al. 2009]), soman (GD), which ages much more quickly (t1/2 = 9 min at 37 °C [Masson et al. 2010]), was used to simulate an aged VX sample. Additional objectives of this exercise included having laboratories assess novel OP-adducted plasma sample preparation techniques and analytical instrumentation methodologies, as well as refining/designating the reporting formats for these new techniques.

  11. Photocatalytic methods for preparation of electrocatalyst materials (United States)

    Li, Wen; Kawamura, Tetsuo; Nagami, Tetsuo; Takahashi, Hiroaki; Muldoon, John; Shelnutt, John A; Song, Yujiang; Miller, James E; Hickner, Michael A; Medforth, Craig


    The invention relates to methods of preparing metal particles on a support material, including platinum-containing nanoparticles on a carbon support. Such materials can be used as electrocatalysts, for example as improved electrocatalysts in polymer electrolyte membrane fuel cells (PEM-FCs).


    NARCIS (Netherlands)

    De Graaf, W.; de Leeuw, J.W.


    Abstract of WO 9412450 (A1) The invention relates to a method of preparing sulfur-containing compounds, comprising reacting a sulfur compound with a compound containing unsaturated carbon-carbon bonds, wherein in a solvent one or more compounds containing non-activated unsaturated carbon-carbon

  13. A robust high-throughput sample preparation and liquid chromatography/tandem mass spectrometry method for the quantitation of β-lyase metabolites of sulfur mustard as 1,1'-sulfonylbis-[2-(methylthio)ethane] in human urine. (United States)

    Reddy, Muntha K; Nixon, Christopher; Wyatt, Shane A; Croley, Timothy R


    Sulfur mustard (HD) is a major chemical warfare agent threat to humans. Since World War I, several incidents of human exposure to sulfur mustard have been reported. In order to assist health professionals during an exposure event and support biological monitoring, a rapid analytical method is required to measure the exposure of humans to HD. The β-lyase metabolites of HD, 1-methylsulfinyl-2-[2-(methylthio)ethylsulfonyl]ethane (MSMTESE) and 1,1'-sulfonylbis[2-(methylsulfinyl)ethane] (SBMSE) were reduced to the single biomarker, 1,1'-sulfonylbis-[2-(methylthio)ethane] (SBMTE), using titanium(III) chloride. High-throughput sample preparation was performed on a Tecan Freedom EVO liquid handler and analysis was performed by electrospray ionization liquid chromatography and tandem mass spectrometry (LC/MS/MS) in the multiple-reaction monitoring mode. Each analytical run consisted of a matrix blank, calibration standards (0.1-100 ng/mL), low quality controls (QCs), 2.5 ng/mL, and high QCs, 25.0 ng/mL, of SBMTE in human urine. The method was validated with 20 analytical runs performed by four analysts. The mean calculated concentrations of the low and high QCs were 2.52 and 25.5 ng/mL with relative standard deviations of 3.6% and 2.3%, respectively. This semi-automated method has few manual transfer steps, thus minimizing common manual errors and saving time. Therefore, this method would be very helpful to responding laboratories in a large-scale exposure event related to HD. Copyright © 2013 John Wiley & Sons, Ltd.

  14. Applications of reversible covalent chemistry in analytical sample preparation. (United States)

    Siegel, David


    Reversible covalent chemistry (RCC) adds another dimension to commonly used sample preparation techniques like solid-phase extraction (SPE), solid-phase microextraction (SPME), molecular imprinted polymers (MIPs) or immuno-affinity cleanup (IAC): chemical selectivity. By selecting analytes according to their covalent reactivity, sample complexity can be reduced significantly, resulting in enhanced analytical performance for low-abundance target analytes. This review gives a comprehensive overview of the applications of RCC in analytical sample preparation. The major reactions covered include reversible boronic ester formation, thiol-disulfide exchange and reversible hydrazone formation, targeting analyte groups like diols (sugars, glycoproteins and glycopeptides, catechols), thiols (cysteinyl-proteins and cysteinyl-peptides) and carbonyls (carbonylated proteins, mycotoxins). Their applications range from low abundance proteomics to reversible protein/peptide labelling to antibody chromatography to quantitative and qualitative food analysis. In discussing the potential of RCC, a special focus is on the conditions and restrictions of the utilized reaction chemistry.

  15. A novel method for preparing microplastic fibers


    Matthew Cole


    Microscopic plastic (microplastic, 0.1??m?5?mm) is a widespread pollutant impacting upon aquatic ecosystems across the globe. Environmental sampling has revealed synthetic fibers are prevalent in seawater, sediments and biota. However, microplastic fibers are rarely used in laboratory studies as they are unavailable for purchase and existing preparation techniques have limited application. To facilitate the incorporation of environmentally relevant microplastic fibers into future studies, new...

  16. A method of language sampling

    DEFF Research Database (Denmark)

    Rijkhoff, Jan; Bakker, Dik; Hengeveld, Kees


    created with this method will reflect optimally the diversity of the languages of the world. On the basis of the internal structure of each genetic language tree a measure is computed that reflects the linguistic diversity in the language families represented by these trees. This measure is used...

  17. Cr(VI) generation during sample preparation of solid samples – A ...

    African Journals Online (AJOL)

    Cr(VI) generation during sample preparation of solid samples – A chromite ore case study. R.I Glastonbury, W van der Merwe, J.P Beukes, P.G van Zyl, G Lachmann, C.J.H Steenkamp, N.F Dawson, M.H Stewart ...

  18. Alpha-spectrometric analysis of uranium and thorium using solid-phase extraction for sample preparation

    Energy Technology Data Exchange (ETDEWEB)

    Weber, R.; Esterlund, R.A.; Patzelt, P


    A method is presented here for the preparation of thin uniform samples of naturally occurring uranium and thorium which are highly suitable for {alpha}-spectrometric analysis. The solid-phase extraction procedure simultaneously achieves a complete separation of the analytes from the sample matrix and a high enrichment factor for uranium and thorium, so that the ensuing eluate is ideally suited for either electrodeposition or ICP-MS, without the need for complicated and painstaking sample preparation. In contrast to conventional liquid-liquid phase-extraction methods, no organic waste solutions are produced, and the process can be easily automated.

  19. Expanding the application of the tablet processing workstation to support the sample preparation of oral suspensions. (United States)

    Opio, Alex Manuel; Nickerson, Beverly; Xue, Gang; Warzeka, John; Norris, Ken


    Sample preparation is the most time-consuming part of the analytical method for powder for oral suspension (POS) assay, purity, and preservative analysis, as this involves multiple dilution and filtration steps. The Tablet Processing Workstation (TPW) was used to automate the sample preparation of a POS formulation. Although the TPW is typically used to automate the preparation of solid oral dosage forms and powders, it contains all of the necessary components to perform POS sample preparation. The TPW exhibited acceptable repeatability in testing 3 lots using 10 replicate preparations per lot. Acceptable linearity of the drug and preservative in the presence of excipients was demonstrated over the range corresponding to 50-150% of intent. Accuracy showed suitable recoveries for all points evaluated. TPW results were shown to correlate to results obtained with the manual method. The TPW method was used to prepare samples in support of manufacturing scale-up efforts. With the efficiencies gained using the TPW, it was possible to analyze a large number of samples generated during process development activities for the POS formulation with minimal human intervention. The extensive data enabled trending of the manufacturing development runs and helped to identify optimization strategies for the process. Copyright © 2011 Society for Laboratory Automation and Screening. Published by Elsevier Inc. All rights reserved.

  20. Preparation Methods: past and Potential Methods of Food Preparation for Space (United States)

    Huber, C. S.


    The logical progression of development of space food systems during the Mercury, Gemini, Apollo, Skylab and Shuttle programs is outlined. The preparation methods which include no preparation to heating, cooling and freezing are reviewed. The introduction of some new and exciting technological advances is proposed, which should result in a system providing crew members with appetizing, safe, nutritious and convenient food.

  1. Highly simplified lateral flow-based nucleic acid sample preparation and passive fluid flow control (United States)

    Cary, Robert E.


    Highly simplified lateral flow chromatographic nucleic acid sample preparation methods, devices, and integrated systems are provided for the efficient concentration of trace samples and the removal of nucleic acid amplification inhibitors. Methods for capturing and reducing inhibitors of nucleic acid amplification reactions, such as humic acid, using polyvinylpyrrolidone treated elements of the lateral flow device are also provided. Further provided are passive fluid control methods and systems for use in lateral flow assays.

  2. New prior sampling methods for nested sampling - Development and testing (United States)

    Stokes, Barrie; Tuyl, Frank; Hudson, Irene


    Nested Sampling is a powerful algorithm for fitting models to data in the Bayesian setting, introduced by Skilling [1]. The nested sampling algorithm proceeds by carrying out a series of compressive steps, involving successively nested iso-likelihood boundaries, starting with the full prior distribution of the problem parameters. The "central problem" of nested sampling is to draw at each step a sample from the prior distribution whose likelihood is greater than the current likelihood threshold, i.e., a sample falling inside the current likelihood-restricted region. For both flat and informative priors this ultimately requires uniform sampling restricted to the likelihood-restricted region. We present two new methods of carrying out this sampling step, and illustrate their use with the lighthouse problem [2], a bivariate likelihood used by Gregory [3] and a trivariate Gaussian mixture likelihood. All the algorithm development and testing reported here has been done with Mathematica® [4].

  3. Combining Electrochemical Sensors with Miniaturized Sample Preparation for Rapid Detection in Clinical Samples

    Directory of Open Access Journals (Sweden)

    Natinan Bunyakul


    Full Text Available Clinical analyses benefit world-wide from rapid and reliable diagnostics tests. New tests are sought with greatest demand not only for new analytes, but also to reduce costs, complexity and lengthy analysis times of current techniques. Among the myriad of possibilities available today to develop new test systems, amperometric biosensors are prominent players—best represented by the ubiquitous amperometric-based glucose sensors. Electrochemical approaches in general require little and often enough only simple hardware components, are rugged and yet provide low limits of detection. They thus offer many of the desirable attributes for point-of-care/point-of-need tests. This review focuses on investigating the important integration of sample preparation with (primarily electrochemical biosensors. Sample clean up requirements, miniaturized sample preparation strategies, and their potential integration with sensors will be discussed, focusing on clinical sample analyses.

  4. GraFix: sample preparation for single-particle electron cryomicroscopy. (United States)

    Kastner, Berthold; Fischer, Niels; Golas, Monika Mariola; Sander, Bjoern; Dube, Prakash; Boehringer, Daniel; Hartmuth, Klaus; Deckert, Jochen; Hauer, Florian; Wolf, Elmar; Uchtenhagen, Hannes; Urlaub, Henning; Herzog, Franz; Peters, Jan Michael; Poerschke, Dietmar; Lührmann, Reinhard; Stark, Holger


    We developed a method, named GraFix, that considerably improves sample quality for structure determination by single-particle electron cryomicroscopy (cryo-EM). GraFix uses a glycerol gradient centrifugation step in which the complexes are centrifuged into an increasing concentration of a chemical fixation reagent to prevent aggregation and to stabilize individual macromolecules. The method can be used to prepare samples for negative-stain, cryo-negative-stain and, particularly, unstained cryo-EM.

  5. Compositions, antibodies, asthma diagnosis methods, and methods for preparing antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Hongjun; Zangar, Richard C.


    Methods for preparing an antibody are provided with the method including incorporating 3-bromo-4-hydroxy-benzoic acid into a protein to form an antigen, immunizing a mammalian host with the antigen, and recovering an antibody having an affinity for the antigen from the host. Antibodies having a binding affinity for a monohalotyrosine are provided as well as composition comprising an antibody bound with monohalotyrosine. Compositions comprising a protein having a 3-bromo-4-hydroxy-benzoic acid moiety are also provided. Methods for evaluating the severity of asthma are provide with the methods including analyzing sputum of a patient using an antibody having a binding affinity for monohalotyrosine, and measuring the amount of antibody bound to protein. Methods for determining eosinophil activity in bodily fluid are also provided with the methods including exposing bodily fluid to an antibody having a binding affinity for monohalotyrosine, and measuring the amount of bound antibody to determine the eosinophil activity.

  6. Protocols for the analytical characterization of therapeutic monoclonal antibodies. II - Enzymatic and chemical sample preparation. (United States)

    Bobaly, Balazs; D'Atri, Valentina; Goyon, Alexandre; Colas, Olivier; Beck, Alain; Fekete, Szabolcs; Guillarme, Davy


    The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. These approaches make the sample more suitable for chromatographic and mass spectrometric analysis. Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. In this paper, general sample preparation approaches used to attain peptide, subunit and glycan level analysis are overviewed. Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). The described protocols may help analysts to develop sample preparation methods in the field of therapeutic protein analysis. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Sample normalization methods in quantitative metabolomics. (United States)

    Wu, Yiman; Li, Liang


    To reveal metabolomic changes caused by a biological event in quantitative metabolomics, it is critical to use an analytical tool that can perform accurate and precise quantification to examine the true concentration differences of individual metabolites found in different samples. A number of steps are involved in metabolomic analysis including pre-analytical work (e.g., sample collection and storage), analytical work (e.g., sample analysis) and data analysis (e.g., feature extraction and quantification). Each one of them can influence the quantitative results significantly and thus should be performed with great care. Among them, the total sample amount or concentration of metabolites can be significantly different from one sample to another. Thus, it is critical to reduce or eliminate the effect of total sample amount variation on quantification of individual metabolites. In this review, we describe the importance of sample normalization in the analytical workflow with a focus on mass spectrometry (MS)-based platforms, discuss a number of methods recently reported in the literature and comment on their applicability in real world metabolomics applications. Sample normalization has been sometimes ignored in metabolomics, partially due to the lack of a convenient means of performing sample normalization. We show that several methods are now available and sample normalization should be performed in quantitative metabolomics where the analyzed samples have significant variations in total sample amounts. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Development of a preparation method to produce a single sample that can be applied to both LC-MS/MS and GC-MS for the screening of postmortem specimens. (United States)

    Hara, Kenji; Waters, Brian; Ikematsu, Natsuki; Tokuyasu, Tomoko; Fujii, Hiroshi; Takayama, Mio; Matsusue, Aya; Kashiwagi, Masayuki; Kubo, Shin-Ichi


    Simple and efficient extraction methods have been developed for the screening of a wide array of drugs in postmortem autopsy specimens. Acidic and basic compounds were targeted with two extraction methods that can be applied to both GC-MS and LC-MS/MS instrumentation. The extractions were achieved by utilizing lipid-removal and solid-phase extraction cartridges while carefully monitoring the pH of the samples to ensure the adequate removal of interfering substances like lipids and amino acid derivatives. These methods were applied to actual autopsy cases, with 94 and 124 compounds detected by GC-MS and LC-MS/MS, respectively. The developed methods could easily be incorporated into a forensic laboratory's daily routine for screening many different compounds from postmortem samples. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  9. A new way of solid dosage form samples preparation for SEM and FTIR using microtome. (United States)

    Šimek, Michal; Grünwaldová, Veronika; Kratochvíl, Bohumil


    Rapid and correct production of generic solid dosage forms requires a large amount of analytical data and conclusions. Modern analytical techniques have a good resolution and accuracy and allow obtaining a lot of information about the original product. Scanning electron microscopy (SEM) is used for observation and assessing individual layers, core and surface of solid dosage forms. Fourier transform infrared (FTIR) spectroscopy mapping allows determining the distribution and characterization of individual components in a solid dosage form. However, the samples prepared by common way, using scalpel or tablet splitter, are not good enough. It was the reason for development of a new and better method of sample preparation, which uses microtome. Well-prepared samples analyzed by SEM and FTIR mapping allow to determine a solid dosage form formulation, excipient content and distribution of excipient and active pharmaceutical ingredient.

  10. The role of sample preparation in interpretation of trace element concentration variability in moss bioindication studies (United States)

    Migaszewski, Z.M.; Lamothe, P.J.; Crock, J.G.; Galuszka, A.; Dolegowska, S.


    Trace element concentrations in plant bioindicators are often determined to assess the quality of the environment. Instrumental methods used for trace element determination require digestion of samples. There are different methods of sample preparation for trace element analysis, and the selection of the best method should be fitted for the purpose of a study. Our hypothesis is that the method of sample preparation is important for interpretation of the results. Here we compare the results of 36 element determinations performed by ICP-MS on ashed and on acid-digested (HNO3, H2O2) samples of two moss species (Hylocomium splendens and Pleurozium schreberi) collected in Alaska and in south-central Poland. We found that dry ashing of the moss samples prior to analysis resulted in considerably lower detection limits of all the elements examined. We also show that this sample preparation technique facilitated the determination of interregional and interspecies differences in the chemistry of trace elements. Compared to the Polish mosses, the Alaskan mosses displayed more positive correlations of the major rock-forming elements with ash content, reflecting those elements' geogenic origin. Of the two moss species, P. schreberi from both Alaska and Poland was also highlighted by a larger number of positive element pair correlations. The cluster analysis suggests that the more uniform element distribution pattern of the Polish mosses primarily reflects regional air pollution sources. Our study has shown that the method of sample preparation is an important factor in statistical interpretation of the results of trace element determinations. ?? 2010 Springer-Verlag.

  11. Dynamic Method for Identifying Collected Sample Mass (United States)

    Carson, John


    G-Sample is designed for sample collection missions to identify the presence and quantity of sample material gathered by spacecraft equipped with end effectors. The software method uses a maximum-likelihood estimator to identify the collected sample's mass based on onboard force-sensor measurements, thruster firings, and a dynamics model of the spacecraft. This makes sample mass identification a computation rather than a process requiring additional hardware. Simulation examples of G-Sample are provided for spacecraft model configurations with a sample collection device mounted on the end of an extended boom. In the absence of thrust knowledge errors, the results indicate that G-Sample can identify the amount of collected sample mass to within 10 grams (with 95-percent confidence) by using a force sensor with a noise and quantization floor of 50 micrometers. These results hold even in the presence of realistic parametric uncertainty in actual spacecraft inertia, center-of-mass offset, and first flexibility modes. Thrust profile knowledge is shown to be a dominant sensitivity for G-Sample, entering in a nearly one-to-one relationship with the final mass estimation error. This means thrust profiles should be well characterized with onboard accelerometers prior to sample collection. An overall sample-mass estimation error budget has been developed to approximate the effect of model uncertainty, sensor noise, data rate, and thrust profile error on the expected estimate of collected sample mass.


    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, S; Brian Culligan, B


    The Savannah River Site Environmental Bioassay Lab participated in the 2007 NRIP Emergency Response program administered by the National Institute for Standards and Technology (NIST) in May, 2007. A new rapid column separation method was applied directly to the NRIP 2007 emergency urine samples, with only minimal sample preparation to reduce preparation time. Calcium phosphate precipitation, previously used to pre-concentrate actinides and Sr-90 in NRIP 2006 urine and water samples, was not used for the NRIP 2007 urine samples. Instead, the raw urine was acidified and passed directly through the stacked resin columns (TEVA+TRU+SR Resins) to separate the actinides and strontium from the NRIP urine samples more quickly. This improvement reduced sample preparation time for the NRIP 2007 emergency urine analyses significantly. This approach works well for small volume urine samples expected during an emergency response event. Based on initial feedback from NIST, the SRS Environmental Bioassay Lab had the most rapid analysis times for actinides and strontium-90 analyses for NRIP 2007 urine samples.

  13. Comparison of sample preparation techniques for large-scale proteomics. (United States)

    Kuljanin, Miljan; Dieters-Castator, Dylan Z; Hess, David A; Postovit, Lynne-Marie; Lajoie, Gilles A


    Numerous workflows exist for large-scale bottom-up proteomics, many of which achieve exceptional proteome depth. Herein, we evaluated the performance of several commonly used sample preparation techniques for proteomic characterization of HeLa lysates [unfractionated in-solution digests, SDS-PAGE coupled with in-gel digestion, gel-eluted liquid fraction entrapment electrophoresis (GELFrEE) technology, SCX StageTips and high-/low-pH reversed phase fractionation (HpH)]. HpH fractionation was found to be superior in terms of proteome depth (>8400 proteins detected) and fractionation efficiency compared to other techniques. SCX StageTip fractionation required minimal sample handling and was also a substantial improvement over SDS-PAGE separation and GELFrEE technology. Sequence coverage of the HeLa proteome increased to 38% when combining all workflows, however, total proteins detected improved only slightly to 8710. In summary, HpH fractionation and SCX StageTips are robust techniques and highly suited for complex proteome analysis. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Automated digital microfluidic sample preparation for next-generation DNA sequencing. (United States)

    Kim, Hanyoup; Bartsch, Michael S; Renzi, Ronald F; He, Jim; Van de Vreugde, James L; Claudnic, Mark R; Patel, Kamlesh D


    Next-generation sequencing (NGS) technology is a promising tool for identifying and characterizing unknown pathogens, but its usefulness in time-critical biodefense and public health applications is currently limited by the lack of fast, efficient, and reliable automated DNA sample preparation methods. To address this limitation, we are developing a digital microfluidic (DMF) platform to function as a fluid distribution hub, enabling the integration of multiple subsystem modules into an automated NGS library sample preparation system. A novel capillary interface enables highly repeatable transfer of liquid between the DMF device and the external fluidic modules, allowing both continuous-flow and droplet-based sample manipulations to be performed in one integrated system. Here, we highlight the utility of the DMF hub platform and capillary interface for automating two key operations in the NGS sample preparation workflow. Using an in-line contactless conductivity detector in conjunction with the capillary interface, we demonstrate closed-loop automated fraction collection of target analytes from a continuous-flow sample stream into droplets on the DMF device. Buffer exchange and sample cleanup, the most repeated steps in NGS library preparation, are also demonstrated on the DMF platform using a magnetic bead assay and achieving an average DNA recovery efficiency of 80%±4.8%. Copyright © 2011 Society for Laboratory Automation and Screening. Published by Elsevier Inc. All rights reserved.

  15. Experimental improvements in sample preparation for the track registration technique from dry and solution media

    Energy Technology Data Exchange (ETDEWEB)

    Suarez-Navarro, M.J. [Universidad Politecnica de Madrid (UPM), E.T.S.I de Caminos, Canales y Puertos, Profesor Aranguren s/n, 28040 Madrid (Spain)]. E-mail:; Pujol, Ll. [Centro de Estudios y Experimentacion de Obras Publicas (CEDEX), Alfonso XII, 3, 28014 Madrid (Spain); Gonzalez-Gonzalez, J.A. [Universidad Politecnica de Madrid (UPM), E.T.S.I de Caminos, Canales y Puertos, Profesor Aranguren s/n, 28040 Madrid (Spain)


    This paper describes the sample preparation studies carried out to determine gross alpha activities in waste materials by means of alpha-particle track counting using CR-39 detector. Sample preparation for the track registration technique using evaporation or electroplating methods (also known as conventional 'dry methods') has a number of drawbacks. The distribution of tracks in different areas of the detector surface is non-uniform, so accurate quantitative determinations depend on tedious and time-consuming counting of tracks under an optical microscope. In this paper, we propose the use of tensioactives in sample preparation to achieve uniform track distribution over the entire detector surface, which enables track density to be evaluated by scanning a small representative area. Under our counting conditions, uniform distribution was achieved with 0.2 ml of Teg from a planchetted source. Furthermore, track registration techniques using solution media (also known as the 'wet methods') and conventional 'dry methods' were analysed and compared with the proposed method. The reproducibility of the procedure described in the study was tested by analysing gross alpha activity in two low-level nuclear waste samples at two different laboratories.

  16. Electromembrane extraction as a rapid and selective miniaturized sample preparation technique for biological fluids

    DEFF Research Database (Denmark)

    Gjelstad, Astrid; Pedersen-Bjergaard, Stig; Seip, Knut Fredrik


    of organic solvent, and into an aqueous receiver solution. The extraction is promoted by application of an electrical field, causing electrokinetic migration of the charged analytes. The method has shown to perform excellent clean-up and selectivity from complicated aqueous matrices like biological fluids......This special report discusses the sample preparation method electromembrane extraction, which was introduced in 2006 as a rapid and selective miniaturized extraction method. The extraction principle is based on isolation of charged analytes extracted from an aqueous sample, across a thin film...

  17. Ultrasonic-based membrane aided sample preparation of urine proteomes. (United States)

    Jesus, Jemmyson Romário; Santos, Hugo M; López-Fernández, H; Lodeiro, Carlos; Arruda, Marco Aurélio Zezzi; Capelo, J L


    A new ultrafast ultrasonic-based method for shotgun proteomics as well as label-free protein quantification in urine samples is developed. The method first separates the urine proteins using nitrocellulose-based membranes and then proteins are in-membrane digested using trypsin. The enzymatic digestion process is accelerated from overnight to four minutes using a sonoreactor ultrasonic device. Overall, the sample treatment pipeline comprising protein separation, digestion and identification is done in just 3h. The process is assessed using urine of healthy volunteers. The method shows that male can be differentiated from female using the protein content of urine in a fast, easy and straightforward way. 232 and 226 proteins are identified in urine of male and female, respectively. From this, 162 are common to both genders, whilst 70 are unique to male and 64 to female. From the 162 common proteins, 13 are present at levels statistically different (p minimalism concept as outlined by Halls, as each stage of this analysis is evaluated to minimize the time, cost, sample requirement, reagent consumption, energy requirements and production of waste products. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Protocol: a fast, comprehensive and reproducible one-step extraction method for the rapid preparation of polar and semi-polar metabolites, lipids, proteins, starch and cell wall polymers from a single sample. (United States)

    Salem, Mohamed A; Jüppner, Jessica; Bajdzienko, Krzysztof; Giavalisco, Patrick


    The elucidation of complex biological systems requires integration of multiple molecular parameters. Accordingly, high throughput methods like transcriptomics, proteomics, metabolomics and lipidomics have emerged to provide the tools for successful system-wide investigations. Unfortunately, optimized analysis of different compounds requires specific extraction procedures in combination with specific analytical instrumentation. However, the most efficient extraction protocols often only cover a restricted number of compounds due to the different physico-chemical properties of these biological compounds. Consequently, comprehensive analysis of several molecular components like polar primary metabolites next to lipids or proteins require multiple aliquots to enable the specific extraction procedures required to cover these diverse compound classes. This multi-parallel sample handling of different sample aliquots is therefore not only more sample intensive, it also requires more time and effort to obtain the required extracts. To circumvent large sample amounts, distributed into several aliquots for the comprehensive extraction of most relevant biological compounds, we developed a simple, robust and reproducible two-phase liquid-liquid extraction protocol. This one-step extraction protocol allows for the analysis of polar-, semi-polar and hydrophobic metabolites, next to insoluble or precipitated compounds, including proteins, starch and plant cell wall components, from a single sample. The method is scalable regarding the used sample amounts but also the employed volumes and can be performed in microcentrifuge tubes, enabling high throughput analysis. The obtained fractions are fully compatible with common analytical methods, including spectroscopic, chromatographic and mass spectrometry-based techniques. To document the utility of the described protocol, we used 25 mg of Arabidopsis thaliana rosette leaves for the generation of multi-omics data sets, covering

  19. Method and apparatus for sampling atmospheric mercury (United States)

    Trujillo, Patricio E.; Campbell, Evan E.; Eutsler, Bernard C.


    A method of simultaneously sampling particulate mercury, organic mercurial vapors, and metallic mercury vapor in the working and occupational environment and determining the amount of mercury derived from each such source in the sampled air. A known volume of air is passed through a sampling tube containing a filter for particulate mercury collection, a first adsorber for the selective adsorption of organic mercurial vapors, and a second adsorber for the adsorption of metallic mercury vapor. Carbon black molecular sieves are particularly useful as the selective adsorber for organic mercurial vapors. The amount of mercury adsorbed or collected in each section of the sampling tube is readily quantitatively determined by flameless atomic absorption spectrophotometry.

  20. On-line sample preparation for the determination of riboflavin and flavin mononucleotides in foodstuffs. (United States)

    Greenway, G M; Kometa, N


    An on-line sample preparation method is described for the determination of riboflavin and flavin mononucleotide (FMN) in milk and cereal samples by high-performance liquid chromatography (HPLC) with fluorescence detection. The on-line system consists of microwave extraction followed by dialysis and trace enrichment with a C18 mini-column. Sample preparation was minimal, with milk samples being directly introduced into the system and cereal only needing to be ground prior to analysis. Results were obtained for a range of samples and these were found to be in agreement with the Association of Official Analytical Chemists (AOAC) method and a previously reported HPLC method. Recoveries were between 94 and 106% for a range of different samples and the relative standard deviation for ten samples was in the range 1.2-2.0%. During the microwave extraction all the flavin adenine dinucleotide (FAD) was converted into FMN and 15% of FMN was converted into riboflavin. The full analysis time on the ground samples was about 20 min.

  1. Membrane biofouling characterization: effects of sample preparation procedures on biofilm structure and the microbial community

    KAUST Repository

    Xue, Zheng


    Ensuring the quality and reproducibility of results from biofilm structure and microbial community analysis is essential to membrane biofouling studies. This study evaluated the impacts of three sample preparation factors (ie number of buffer rinses, storage time at 4°C, and DNA extraction method) on the downstream analysis of nitrifying biofilms grown on ultrafiltration membranes. Both rinse and storage affected biofilm structure, as suggested by their strong correlation with total biovolume, biofilm thickness, roughness and the spatial distribution of EPS. Significant variations in DNA yields and microbial community diversity were also observed among samples treated by different rinses, storage and DNA extraction methods. For the tested biofilms, two rinses, no storage and DNA extraction with both mechanical and chemical cell lysis from attached biofilm were the optimal sample preparation procedures for obtaining accurate information about biofilm structure, EPS distribution and the microbial community. © 2014 © 2014 Taylor & Francis.

  2. Integration of Gas Chromatography Mass Spectrometry Methods for Differentiating Ricin Preparation Methods

    Energy Technology Data Exchange (ETDEWEB)

    Wunschel, David S.; Melville, Angela M.; Ehrhardt, Christopher J.; Colburn, Heather A.; Victry, Kristin D.; Antolick, Kathryn C.; Wahl, Jon H.; Wahl, Karen L.


    The investigation of crimes involving chemical or biological agents is infrequent, but presents unique analytical challenges. The protein toxin ricin is encountered more frequently than other agents and is found in the seeds of the castor plant Ricinus communis. Typically, the toxin is extracted from castor seeds utilizing a variety of different recipes that result in varying purity of the toxin. Moreover, these various purification steps can also leave or differentially remove a variety of exogenous and endogenous residual components with the toxin that may indicate the type and number of purification steps involved. We have applied three gas chromatographic - mass spectrometric (GC-MS) based analytical methods to measure the variation in seed carbohydrates and castor oil ricinoleic acid as well as the presence of solvents used for purification. These methods were applied to the same samples prepared using four previously identified toxin preparation methods starting from four varieties of castor seeds. The individual data sets for seed carbohydrate profiles, ricinoleic acid or acetone amount each provided information capable of differentiating different types of toxin preparations across seed types. However, the integration of the data sets using multivariate factor analysis provided a clear distinction of all samples based on the preparation method and independent of the seed source. In particular the abundance of mannose, arabinose, fucose, ricinoleic acid and acetone were shown to be important differentiating factors. These complementary tools provide a more confident determination of the method of toxin preparation.

  3. Sampling of temporal networks: Methods and biases (United States)

    Rocha, Luis E. C.; Masuda, Naoki; Holme, Petter


    Temporal networks have been increasingly used to model a diversity of systems that evolve in time; for example, human contact structures over which dynamic processes such as epidemics take place. A fundamental aspect of real-life networks is that they are sampled within temporal and spatial frames. Furthermore, one might wish to subsample networks to reduce their size for better visualization or to perform computationally intensive simulations. The sampling method may affect the network structure and thus caution is necessary to generalize results based on samples. In this paper, we study four sampling strategies applied to a variety of real-life temporal networks. We quantify the biases generated by each sampling strategy on a number of relevant statistics such as link activity, temporal paths and epidemic spread. We find that some biases are common in a variety of networks and statistics, but one strategy, uniform sampling of nodes, shows improved performance in most scenarios. Given the particularities of temporal network data and the variety of network structures, we recommend that the choice of sampling methods be problem oriented to minimize the potential biases for the specific research questions on hand. Our results help researchers to better design network data collection protocols and to understand the limitations of sampled temporal network data.

  4. Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography. (United States)

    Trépout, Sylvain; Bastin, Philippe; Marco, Sergio


    This report describes a protocol for preparing thick biological specimens for further observation using a scanning transmission electron microscope. It also describes an imaging method for studying the 3D structure of thick biological specimens by scanning transmission electron tomography. The sample preparation protocol is based on conventional methods in which the sample is fixed using chemical agents, treated with a heavy atom salt contrasting agent, dehydrated in a series of ethanol baths, and embedded in resin. The specific imaging conditions for observing thick samples by scanning transmission electron microscopy are then described. Sections of the sample are observed using a through-focus method involving the collection of several images at various focal planes. This enables the recovery of in-focus information at various heights throughout the sample. This particular collection pattern is performed at each tilt angle during tomography data collection. A single image is then generated, merging the in-focus information from all the different focal planes. A classic tilt-series dataset is then generated. The advantage of the method is that the tilt-series alignment and reconstruction can be performed using standard tools. The collection of through-focal images allows the reconstruction of a 3D volume that contains all of the structural details of the sample in focus.

  5. Microsystem strategies for sample preparation in biological detection.

    Energy Technology Data Exchange (ETDEWEB)

    James, Conrad D.; Galambos, Paul C.; Bennett, Dawn Jonita (University of Maryland Baltimore County, Baltimore, MD); Manginell, Monica; Okandan, Murat; Acrivos, Andreas (The City College of New York, NY); Brozik, Susan Marie; Khusid, Boris (New Jersey Institute of Technology, Newark, NJ)


    The objective of this LDRD was to develop microdevice strategies for dealing with samples to be examined in biological detection systems. This includes three sub-components: namely, microdevice fabrication, sample delivery to the microdevice, and sample processing within the microdevice. The first component of this work focused on utilizing Sandia's surface micromachining technology to fabricate small volume (nanoliter) fluidic systems for processing small quantities of biological samples. The next component was to develop interfaces for the surface-micromachined silicon devices. We partnered with Micronics, a commercial company, to produce fluidic manifolds for sample delivery to our silicon devices. Pressure testing was completed to examine the strength of the bond between the pressure-sensitive adhesive layer and the silicon chip. We are also pursuing several other methods, both in house and external, to develop polymer-based fluidic manifolds for packaging silicon-based microfluidic devices. The second component, sample processing, is divided into two sub-tasks: cell collection and cell lysis. Cell collection was achieved using dielectrophoresis, which employs AC fields to collect cells at energized microelectrodes, while rejecting non-cellular particles. Both live and dead Staph. aureus bacteria have been collected using RF frequency dielectrophoresis. Bacteria have been separated from polystyrene microspheres using frequency-shifting dielectrophoresis. Computational modeling was performed to optimize device separation performance, and to predict particle response to the dielectrophoretic traps. Cell lysis is continuing to be pursued using microactuators to mechanically disrupt cell membranes. Novel thermal actuators, which can generate larger forces than previously tested electrostatic actuators, have been incorporated with and tested with cell lysis devices. Significant cell membrane distortion has been observed, but more experiments need to be

  6. A random spatial sampling method in a rural developing nation. (United States)

    Kondo, Michelle C; Bream, Kent D W; Barg, Frances K; Branas, Charles C


    Nonrandom sampling of populations in developing nations has limitations and can inaccurately estimate health phenomena, especially among hard-to-reach populations such as rural residents. However, random sampling of rural populations in developing nations can be challenged by incomplete enumeration of the base population. We describe a stratified random sampling method using geographical information system (GIS) software and global positioning system (GPS) technology for application in a health survey in a rural region of Guatemala, as well as a qualitative study of the enumeration process. This method offers an alternative sampling technique that could reduce opportunities for bias in household selection compared to cluster methods. However, its use is subject to issues surrounding survey preparation, technological limitations and in-the-field household selection. Application of this method in remote areas will raise challenges surrounding the boundary delineation process, use and translation of satellite imagery between GIS and GPS, and household selection at each survey point in varying field conditions. This method favors household selection in denser urban areas and in new residential developments. Random spatial sampling methodology can be used to survey a random sample of population in a remote region of a developing nation. Although this method should be further validated and compared with more established methods to determine its utility in social survey applications, it shows promise for use in developing nations with resource-challenged environments where detailed geographic and human census data are less available.

  7. Platinum adlayered ruthenium nanoparticles, method for preparing, and uses thereof (United States)

    Tong, YuYe; Du, Bingchen


    A superior, industrially scalable one-pot ethylene glycol-based wet chemistry method to prepare platinum-adlayered ruthenium nanoparticles has been developed that offers an exquisite control of the platinum packing density of the adlayers and effectively prevents sintering of the nanoparticles during the deposition process. The wet chemistry based method for the controlled deposition of submonolayer platinum is advantageous in terms of processing and maximizing the use of platinum and can, in principle, be scaled up straightforwardly to an industrial level. The reactivity of the Pt(31)-Ru sample was about 150% higher than that of the industrial benchmark PtRu (1:1) alloy sample but with 3.5 times less platinum loading. Using the Pt(31)-Ru nanoparticles would lower the electrode material cost compared to using the industrial benchmark alloy nanoparticles for direct methanol fuel cell applications.

  8. Sampling methods for terrestrial amphibians and reptiles. (United States)

    Paul Stephen Corn; R. Bruce. Bury


    Methods described for sampling amphibians and reptiles in Douglas-fir forests in the Pacific Northwest include pitfall trapping, time-constrained collecting, and surveys of coarse woody debris. The herpetofauna of this region differ in breeding and nonbreeding habitats and vagility, so that no single technique is sufficient for a community study. A combination of...

  9. Automated sample preparation for ICP analysis of active pharmaceutical ingredients and intermediates. (United States)

    Sims, Jonathan; Smith, Andrew; Patel, Dharmista; Batchelor, Richard; Carreira, Judith


    Routine testing of active pharmaceutical ingredients (APIs) for metal residues is an expectation of regulatory bodies such as the FDA (U.S. Food and Drug Administration). Sample preparation techniques are the rate-limiting step in the testing process and can be variable depending on the specific characteristics of the API under test. Simplification and standardization of the routine preparation of inductively coupled plasma spectroscopy sample solutions of organic compounds has been developed using a commercially available robotic workstation. Contamination from the metal components of the instrument and from sample tubes used in the methodology has been studied using a Design of Experiments approach. The optimized method described can be used for the measurement of trace metals in Pharmaceuticals at levels compliant with European and U.S. regulatory requirements. Copyright © 2011 Society for Laboratory Automation and Screening. Published by Elsevier Inc. All rights reserved.

  10. Yeast metabolomics: sample preparation for a GC/MS-based analysis. (United States)

    Carneiro, Sónia; Pereira, Rui; Rocha, Isabel


    Metabolome sample preparation is one of the key factors in metabolomics analyses. The quality of the metabolome data will depend on the suitability of the experimental procedures to the cellular system (e.g., yeast cells) and the analytical performance. Here, we summarize a protocol for metabolome analysis of yeast cells using gas chromatography-mass spectrometry (GC-MS). First, the main phases of a metabolomics analysis are identified: sample preparation, metabolite extraction, and analysis. We also provide an overview on different methods used to quench samples and extract intracellular metabolites from yeast cells. This protocol provides a detailed description of a GC-MS-based analysis of yeast metabolome, in particular for metabolites containing amino and/or carboxyl groups, which represent most of the compounds participating in the central carbon metabolism.

  11. An On-Target Desalting and Concentration Sample Preparation Protocol for MALDI-MS and MS/MS Analysis

    DEFF Research Database (Denmark)

    Zhang, Xumin; Wang, Quanhui; Lou, Xiaomin


    2DE coupled with MALDI-MS is one of the most widely used and powerful analytic technologies in proteomics study. The MALDI sample preparation method has been developed and optimized towards the combination of simplicity, sample-cleaning, and sample concentration since its introduction. Here we pr...

  12. Bio-sample preparation and gas chromatographic determination of benzodiazepines--a review. (United States)

    Uddin, Mohammad Nasir; Samanidou, Victoria F; Papadoyannis, Ioannis N


    Benzodiazepines have become commonly prescribed medicines worldwide in the therapy of anxiety, sleep disorders and convulsive attacks because they are relatively safe, with mild side effects. The availability of rapid, sensitive and selective analytical methods is essential for the determination of these drugs in clinical and forensic cases. Benzodiazepines are usually present at trace levels (μg/mL or ng/mL) in a complex biological matrix, and the potentially interfering compounds need to be removed before analysis. Therefore, a sample preparation technique is often mandatory, both to extract the drugs of interest from the matrices and to increase their concentration. An extended and comprehensive review is presented herein, focusing on bio-sample preparation (pretreatment, extraction and derivatization) and gas chromatographic methods applied for the quantification of 1,4-benzodiazepines.

  13. [Research on the preparative method of Arctigenin]. (United States)

    Zhang, Li-Ying; Yang, Yi-Shun; Zhang, Tong; Ding, Yue; Cai, Zhen-Zhen; Tao, Jian-Sheng


    To research on the preparation of Arctigenin in vitro. Took enzyme concentration, time course and substrate concentration as investigation factors, used Box-Behnken design-response surface methodology to optimize the enzyme hydrolysis path of Arctigenin. The best operational path for Arctigenin was as follows: the temperature was 50 degrees C, pH was 4.8, enzyme concentration was 0.44 U/mL, time course was 46.81 min, substrate concentration was 0.29 mg/mL, the conversion rate was 90.94%. This research can be regarded as a referencein preparing Arctigenin in vitro.

  14. Standardized Sample Preparation Using a Drop-on-Demand Printing Platform (United States)


    ring” effect , this effect is statistically negligible as compared to drop-and-dry methods. This is of particular concern with spatially sensitive ...Gillen, J.G. Fabrication of polymer microsphere particle standards containing trace explosives using an oil/water emulsion solvent extraction...technique and sample preparation protocol for explosive materials testing based on drop-on-demand technology. Droplet mass and reproducibility were measured

  15. Review of sample preparation techniques for the analysis of pesticide residues in soil. (United States)

    Tadeo, José L; Pérez, Rosa Ana; Albero, Beatriz; García-Valcárcel, Ana I; Sánchez-Brunete, Consuelo


    This paper reviews the sample preparation techniques used for the analysis of pesticides in soil. The present status and recent advances made during the last 5 years in these methods are discussed. The analysis of pesticide residues in soil requires the extraction of analytes from this matrix, followed by a cleanup procedure, when necessary, prior to their instrumental determination. The optimization of sample preparation is a very important part of the method development that can reduce the analysis time, the amount of solvent, and the size of samples. This review considers all aspects of sample preparation, including extraction and cleanup. Classical extraction techniques, such as shaking, Soxhlet, and ultrasonic-assisted extraction, and modern techniques like pressurized liquid extraction, microwave-assisted extraction, solid-phase microextraction and QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) are reviewed. The different cleanup strategies applied for the purification of soil extracts are also discussed. In addition, the application of these techniques to environmental studies is considered.

  16. Preparation and characterisation of magnetic nanostructured samples for inelastic neutron scattering experiments

    Energy Technology Data Exchange (ETDEWEB)

    Kreuzpaintner, Wolfgang


    Recent advances in thin-film structuring techniques have generated significant interest in the dynamics of spin waves in magnetic nanostructures and the possible use of inelastic neutron scattering (INS) for their investigation. This thesis describes the design and implementation, at GKSS Research Centre, of equipment for preparation of large and laterally submicron and nanometre structured magnetic samples for such future INS experiments. After a brief resume on spin waves in nanostructures, the development work on new purpose-designed equipment, including high vacuum (HV) argon ion beam milling and ultra high vacuum (UHV) e-beam evaporation setups, is described. Ni nanodot as well as Ni and novel Gd nanowire samples were prepared using combinations of sputter deposition, laser interference lithography, argon ion beam milling, e-beam evaporation and self organisation techniques. With reference to sample preparation, epitaxial growth studies for Ni on Si(100) substrate were performed, resulting in the development of a new deposition process, which by thermal tuning allows for the direct epitaxial growth of Ni on Si with unprecedented crystalline quality. The results of various characterisation experiments on the prepared nanostructured samples, including Scanning Electron Microscopy (SEM), microprobe analysis, Atomic and Magnetic Force Microscopy (AFM/MFM), Vibrating Sample Magnetometry (VSM), X-ray Diffraction (XRD) and Reflectivity (XRR), unpolarised and Polarised Neutron Scattering (PNR) and off-specular scattering by X-rays and neutrons using rocking scans and Time-Of-Flight Grazing Incidence Small Angle Neutron Scattering (TOF-GISANS), together with various analysis procedures such as Distorted-Wave Born Approximation (DWBA), are reported. The analysis of a Gd nanowire sample by TOF-GISANS led to a novel evaluation technique which in comparison with single wavelength methods allows portions of reciprocal space to be scanned without changing the angle of

  17. Gas-Assisted Annular Microsprayer for Sample Preparation for Time-Resolved Cryo-Electron Microscopy. (United States)

    Lu, Zonghuan; Barnard, David; Shaikh, Tanvir R; Meng, Xing; Mannella, Carmen A; Yassin, Aymen; Agrawal, Rajendra; Wagenknecht, Terence; Lu, Toh-Ming


    Time-resolved cryo electron microscopy (TRCEM) has emerged as a powerful technique for transient structural characterization of isolated biomacromolecular complexes in their native state within the time scale of seconds to milliseconds. For TRCEM sample preparation, microfluidic device [9] has been demonstrated to be a promising approach to facilitate TRCEM biological sample preparation. It is capable of achieving rapidly aqueous sample mixing, controlled reaction incubation, and sample deposition on electron microscopy (EM) grids for rapid freezing. One of the critical challenges is to transfer samples to cryo-EM grids from the microfluidic device. By using microspraying method, the generated droplet size needs to be controlled to facilitate the thin ice film formation on the grid surface for efficient data collection, while not too thin to be dried out before freezing, i.e., optimized mean droplet size needs to be achieved. In this work, we developed a novel monolithic three dimensional (3D) annular gas-assisted microfluidic sprayer using 3D MEMS (MicroElectroMechanical System) fabrication techniques. The microsprayer demonstrated dense and consistent microsprays with average droplet size between 6-9 μm, which fulfilled the above droplet size requirement for TRCEM sample preparation. With droplet density of around 12-18 per grid window (window size is 58×58 μm), and the data collectible thin ice region of >50% total wetted area, we collected ~800-1000 high quality CCD micrographs in a 6-8 hour period of continuous effort. This level of output is comparable to what were routinely achieved using cryo-grids prepared by conventional blotting and manual data collection. In this case, weeks of data collection process with the previous device [9] has shortened to a day or two. And hundreds of microliter of valuable sample consumption can be reduced to only a small fraction.

  18. Preparation of Plant 41Ca Tracer Samples for Accelerator Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    ZHAO Qing-zhang1;JANG Ping-ping3;LIN De-yu4;YANG Xian-lin1;DOU Liang1;PANG Yi-jun1;WANG Xiao-ming1;ZHANG Hui1,5;YANG Xu-ran1;WU Shao-yong1;GAO Dong-sheng2;LI Ling2;WANG Lei2;SUN Ke-peng2;ZHOU Jun2;DONG Ke-jun1;HE Ming1


    Full Text Available Calcium plays an important role in the metabolism of plants and animals. In this paper, the preparation method of plant 41Ca for accelerator mass spectrometry (AMS measurement was developed for the first time in China. AMS, with its advantages of high sensitivity, small dose of radioactivity, high accuracy, large measuring range, and long tracer cycle, can be used to measure cosmogenic nuclide 41Ca , which has long half-life. The intensity of the beam in ion source is an important parameter for the sensitivity of AMS measurement. The high beam current can improve the sensitivity of AMS. The preparation methods of plant samples of 41Ca tracer were systematically studied to obtain high beam current using wet, dry and a combining method with wet and dry re-fluoride. A reliable preparation procedure of plant samples for 41Ca tracer and its optimization parameters were determined by testing beam currents of various samples and lay a foundation for the 41Ca-AMS technology at plant tracer applications.

  19. Evaluation of Sampling and Sample Preparation Modifications for Soil Containing Metallic Residues (United States)


    representation of the overall experimental design. Soil samples were transported to CRREL and air-dried on alumi - num trays. Once air-dried, each...60 s and then spread out onto an alumi - num cookie sheet. Digestion generally followed USEPA Method 3050B with the following exceptions. Two grams of

  20. HPLC/DAD determination of rosmarinic acid in Salvia officinalis: sample preparation optimization by factorial design

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Karina B. de [Universidade Federal do Parana (UFPR), Curitiba, PR (Brazil). Dept. de Farmacia; Oliveira, Bras H. de, E-mail: [Universidade Federal do Parana (UFPR), Curitiba, PR (Brazil). Dept. de Quimica


    Sage (Salvia officinalis) contains high amounts of the biologically active rosmarinic acid (RA) and other polyphenolic compounds. RA is easily oxidized, and may undergo degradation during sample preparation for analysis. The objective of this work was to develop and validate an analytical procedure for determination of RA in sage, using factorial design of experiments for optimizing sample preparation. The statistically significant variables for improving RA extraction yield were determined initially and then used in the optimization step, using central composite design (CCD). The analytical method was then fully validated, and used for the analysis of commercial samples of sage. The optimized procedure involved extraction with aqueous methanol (40%) containing an antioxidant mixture (ascorbic acid and ethylenediaminetetraacetic acid (EDTA)), with sonication at 45 deg C for 20 min. The samples were then injected in a system containing a C{sub 18} column, using methanol (A) and 0.1% phosphoric acid in water (B) in step gradient mode (45A:55B, 0-5 min; 80A:20B, 5-10 min) with flow rate of 1.0 mL min-1 and detection at 330 nm. Using this conditions, RA concentrations were 50% higher when compared to extractions without antioxidants (98.94 {+-} 1.07% recovery). Auto-oxidation of RA during sample extraction was prevented by the use of antioxidants resulting in more reliable analytical results. The method was then used for the analysis of commercial samples of sage. (author)

  1. Fast detection of Noroviruses using a real-time PCR assay and automated sample preparation

    Directory of Open Access Journals (Sweden)

    Schmid Michael


    Full Text Available Abstract Background Noroviruses (NoV have become one of the most commonly reported causative agents of large outbreaks of non-bacterial acute gastroenteritis worldwide as well as sporadic gastroenteritis in the community. Currently, reverse transcriptase polymerase chain reaction (RT-PCR assays have been implemented in NoV diagnosis, but improvements that simplify and standardize sample preparation, amplification, and detection will be further needed. The combination of automated sample preparation and real-time PCR offers such refinements. Methods We have designed a new real-time RT-PCR assay on the LightCycler (LC with SYBR Green detection and melting curve analysis (Tm to detect NoV RNA in patient stool samples. The performance of the real-time PCR assay was compared with that obtained in parallel with a commercially available enzyme immunoassay (ELISA for antigen detection by testing a panel of 52 stool samples. Additionally, in a collaborative study with the Baden-Wuerttemberg State Health office, Stuttgart (Germany the real-time PCR results were blindly assessed using a previously well-established nested PCR (nPCR as the reference method, since PCR-based techniques are now considered as the "gold standard" for NoV detection in stool specimens. Results Analysis of 52 clinical stool samples by real-time PCR yielded results that were consistent with reference nPCR results, while marked differences between the two PCR-based methods and antigen ELISA were observed. Our results indicate that PCR-based procedures are more sensitive and specific than antigen ELISA for detecting NoV in stool specimens. Conclusions The combination of automated sample preparation and real-time PCR provided reliable diagnostic results in less time than conventional RT-PCR assays. These benefits make it a valuable tool for routine laboratory practice especially in terms of rapid and appropriate outbreak-control measures in health-care facilities and other settings.

  2. Challenges of biological sample preparation for SIMS imaging of elements and molecules at subcellular resolution (United States)

    Chandra, Subhash


    Secondary ion mass spectrometry (SIMS) based imaging techniques capable of subcellular resolution characterization of elements and molecules are becoming valuable tools in many areas of biology and medicine. Due to high vacuum requirements of SIMS, the live cells cannot be analyzed directly in the instrument. The sample preparation, therefore, plays a critical role in preserving the native chemical composition for SIMS analysis. This work focuses on the evaluation of frozen-hydrated and frozen freeze-dried sample preparations for SIMS studies of cultured cells with a CAMECA IMS-3f dynamic SIMS ion microscope instrument capable of producing SIMS images with a spatial resolution of 500 nm. The sandwich freeze-fracture method was used for fracturing the cells. The complimentary fracture planes in the plasma membrane were characterized by field-emission secondary electron microscopy (FESEM) in the frozen-hydrated state. The cells fractured at the dorsal surface were used for SIMS analysis. The frozen-hydrated SIMS analysis of individual cells under dynamic primary ion beam (O 2+) revealed local secondary ion signal enhancements correlated with the water image signals of 19(H 3O) +. A preferential removal of water from the frozen cell matrix in the Z-axis was also observed. These complications render the frozen-hydrated sample type less desirable for subcellular dynamic SIMS studies. The freeze-drying of frozen-hydrated cells, either inside the instrument or externally in a freeze-drier, allowed SIMS imaging of subcellular chemical composition. Morphological evaluations of fractured freeze-dried cells with SEM and confocal laser scanning microscopy (CLSM) revealed well-preserved mitochondria, Golgi apparatus, and stress fibers. SIMS analysis of fractured freeze-dried cells revealed well-preserved chemical composition of even the most highly diffusible ions like K + and Na + in physiologically relevant concentrations. The high K-low Na signature in individual cells

  3. Challenges of biological sample preparation for SIMS imaging of elements and molecules at subcellular resolution

    Energy Technology Data Exchange (ETDEWEB)

    Chandra, Subhash [Cornell SIMS Laboratory, Department of Earth and Atmospheric Sciences, Snee Hall, Cornell University, Ithaca, NY 14853 (United States)], E-mail:


    Secondary ion mass spectrometry (SIMS) based imaging techniques capable of subcellular resolution characterization of elements and molecules are becoming valuable tools in many areas of biology and medicine. Due to high vacuum requirements of SIMS, the live cells cannot be analyzed directly in the instrument. The sample preparation, therefore, plays a critical role in preserving the native chemical composition for SIMS analysis. This work focuses on the evaluation of frozen-hydrated and frozen freeze-dried sample preparations for SIMS studies of cultured cells with a CAMECA IMS-3f dynamic SIMS ion microscope instrument capable of producing SIMS images with a spatial resolution of 500 nm. The sandwich freeze-fracture method was used for fracturing the cells. The complimentary fracture planes in the plasma membrane were characterized by field-emission secondary electron microscopy (FESEM) in the frozen-hydrated state. The cells fractured at the dorsal surface were used for SIMS analysis. The frozen-hydrated SIMS analysis of individual cells under dynamic primary ion beam (O{sub 2}{sup +}) revealed local secondary ion signal enhancements correlated with the water image signals of {sup 19}(H{sub 3}O){sup +}. A preferential removal of water from the frozen cell matrix in the Z-axis was also observed. These complications render the frozen-hydrated sample type less desirable for subcellular dynamic SIMS studies. The freeze-drying of frozen-hydrated cells, either inside the instrument or externally in a freeze-drier, allowed SIMS imaging of subcellular chemical composition. Morphological evaluations of fractured freeze-dried cells with SEM and confocal laser scanning microscopy (CLSM) revealed well-preserved mitochondria, Golgi apparatus, and stress fibers. SIMS analysis of fractured freeze-dried cells revealed well-preserved chemical composition of even the most highly diffusible ions like K{sup +} and Na{sup +} in physiologically relevant concentrations. The high K

  4. Comparison of sampling methods for radiocarbon dating of carbonyls in air samples via accelerator mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Schindler, Matthias, E-mail:; Kretschmer, Wolfgang; Scharf, Andreas; Tschekalinskij, Alexander


    Three new methods to sample and prepare various carbonyl compounds for radiocarbon measurements were developed and tested. Two of these procedures utilized the Strecker synthetic method to form amino acids from carbonyl compounds with either sodium cyanide or trimethylsilyl cyanide. The third procedure used semicarbazide to form crystalline carbazones with the carbonyl compounds. The resulting amino acids and semicarbazones were then separated and purified using thin layer chromatography. The separated compounds were then combusted to CO{sub 2} and reduced to graphite to determine {sup 14}C content by accelerator mass spectrometry (AMS). All of these methods were also compared with the standard carbonyl compound sampling method wherein a compound is derivatized with 2,4-dinitrophenylhydrazine and then separated by high-performance liquid chromatography (HPLC).

  5. Filter-Aided Sample Preparation (FASP) for Improved Proteome Analysis of Recombinant Chinese Hamster Ovary Cells. (United States)

    Coleman, Orla; Henry, Michael; Clynes, Martin; Meleady, Paula


    Chinese hamster ovary (CHO) cells are the most commonly used mammalian host cell line for biopharmaceutical production because of their ability to correctly fold and posttranslationally modify recombinant proteins that are compatible with human use. Proteomics, along with other 'omic platforms, are being used to understand the biology of CHO cells with the ultimate aim of enhancing CHO cell factories for more efficient production of biopharmaceuticals. In this chapter, we will describe an efficient protocol called Filter Aided Sample Preparation (FASP) for the extraction of proteins from CHO cells for proteomic studies. FASP uses a common ultrafiltration device whereby the membrane pores are small enough to allow contaminating detergents to pass through, while proteins are too large and are retained and concentrated in the filter unit. This method of sample preparation and protein digestion is universally applicable and can be easily employed in any proteomics facilities as standard everyday laboratory reagents and equipment are used.

  6. Molecularly imprinted polymers for sample preparation and biosensing in food analysis: Progress and perspectives

    DEFF Research Database (Denmark)

    Ashley, Jon; Shahbazi, Mohammad-Ali; Kant, Krishna


    Molecularly imprinted polymers (MIPs) are biomimetics which can selectively bind to analytes of interest. One of the most interesting areas where MIPs have shown the biggest potential is food analysis. MIPs have found use as sorbents in sample preparation attributed to the high selectivity and high....... MIPs have been utilized as receptors in biosensing platforms such as electrochemical, optical and mass biosensors to detect various analytes in food. In this review, we will discuss the current state-of-the-art of MIP synthesis and applications in the context of food analysis. We will highlight...... the imprinting methods which are applicable for imprinting food templates, summarize the recent progress in using MIPs for preparing and analysing food samples, and discuss the current limitations in the commercialisation of MIPs technology. Finally, future perspectives will be given....

  7. FISHprep: A Novel Integrated Device for Metaphase FISH Sample Preparation

    DEFF Research Database (Denmark)

    Shah, Pranjul Jaykumar; Vedarethinam, Indumathi; Kwasny, Dorota


    a novel device with an integrated expansion chamber to culture, arrest and fix metaphase cells followed by a subsequent splashing protocol leading to ample metaphase chromosome spreads on a glass slide for metaphase FISH analysis. The device provides an easy, disposable, low cost, integrated solution...... with minimal handling for metaphase FISH slide preparation....

  8. Minimizing technical variation during sample preparation prior to label-free quantitative mass spectrometry. (United States)

    Scheerlinck, E; Dhaenens, M; Van Soom, A; Peelman, L; De Sutter, P; Van Steendam, K; Deforce, D


    Sample preparation is the crucial starting point to obtain high-quality mass spectrometry data and can be divided into two main steps in a bottom-up proteomics approach: cell/tissue lysis with or without detergents and a(n) (in-solution) digest comprising denaturation, reduction, alkylation, and digesting of the proteins. Here, some important considerations, among others, are that the reagents used for sample preparation can inhibit the digestion enzyme (e.g., 0.1% sodium dodecyl sulfate [SDS] and 0.5 M guanidine HCl), give rise to ion suppression (e.g., polyethylene glycol [PEG]), be incompatible with liquid chromatography-tandem mass spectrometry (LC-MS/MS) (e.g., SDS), and can induce additional modifications (e.g., urea). Taken together, all of these irreproducible effects are gradually becoming a problem when label-free quantitation of the samples is envisioned such as during the increasingly popular high-definition mass spectrometry (HDMS(E)) and sequential window acquisition of all theoretical fragment ion spectra (SWATH) data-independent acquisition strategies. Here, we describe the detailed validation of a reproducible method with sufficient protein yield for sample preparation without any known LC-MS/MS interfering substances by using 1% sodium deoxycholate (SDC) during both cell lysis and in-solution digest. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  9. Molecularly imprinted polymers for sample preparation and biosensing in food analysis: Progress and perspectives. (United States)

    Ashley, Jon; Shahbazi, Mohammad-Ali; Kant, Krishna; Chidambara, Vinayaka Aaydha; Wolff, Anders; Bang, Dang Duong; Sun, Yi


    Molecularly imprinted polymers (MIPs) are biomimetics which can selectively bind to analytes of interest. One of the most interesting areas where MIPs have shown the biggest potential is food analysis. MIPs have found use as sorbents in sample preparation attributed to the high selectivity and high loading capacity. MIPs have been intensively employed in classical solid-phase extraction and solid-phase microextraction. More recently, MIPs have been combined with magnetic bead extraction, which greatly simplifies sample handling procedures. Studies have consistently shown that MIPs can effectively minimize complex food matrix effects, and improve recoveries and detection limits. In addition to sample preparation, MIPs have also been viewed as promising alternatives to bio-receptors due to the inherent molecular recognition abilities and the high stability in harsh chemical and physical conditions. MIPs have been utilized as receptors in biosensing platforms such as electrochemical, optical and mass biosensors to detect various analytes in food. In this review, we will discuss the current state-of-the-art of MIP synthesis and applications in the context of food analysis. We will highlight the imprinting methods which are applicable for imprinting food templates, summarize the recent progress in using MIPs for preparing and analysing food samples, and discuss the current limitations in the commercialisation of MIPs technology. Finally, future perspectives will be given. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Sample preparation for liquid chromatographic analysis of phytochemicals in biological fluids. (United States)

    Oh, Ju-Hee; Lee, Young-Joo


    Natural products have been used traditionally for the treatment and prevention of diseases for thousands of years and are nowadays consumed as dietary supplements and herbal medicine. To ensure the safe and effective use of these herbal products, information about bioavailability of active compounds in plasma or target tissues should be provided via validated analytical methods combined with appropriate sampling methods. To provide comprehensive and abridged information about sample preparation methods for the quantification of phytochemicals in biological samples using liquid chromatography analysis. Sample pre-treatment procedures used in analytical methods for in vivo pharmacokinetic studies of natural compounds or herbal medicines were reviewed. These were categorised according to the biological matrices (plasma, bile, urine, faeces and tissues) and sample clean-up processes (protein precipitation, liquid-liquid extraction and solid-phase extraction). Although various kinds of sample pre-treatment methods have been developed, liquid-liquid extraction is still widely used and solid-phase extraction is becoming increasingly popular because of its efficiency for extensive clean up of complex matrix samples. However, protein precipitation is still favoured due to its simplicity. Sample treatment for phytochemical analysis in biological fluids is an indispensable and critical step to obtain high quality results. This step could dominate the overall analytical process because both the duration of the process as well as the reliability of the data depend in large part on its efficiency. Thus, special attention should be given to the choice of a proper sample treatment method that targets analytes and their biomatrix. Copyright © 2013 John Wiley & Sons, Ltd.

  11. Resin catalysts and method of preparation (United States)

    Smith, L.A. Jr.


    Heat stabilized catalyst compositions are prepared from nuclear sulfonic acid, for example, macroporous crosslinked polyvinyl aromatic compounds containing sulfonic acid groups are neutralized with a metal of Al, Fe, Zn, Cu, Ni, ions or mixtures and alkali, alkaline earth metals or ammonium ions by contacting the resin containing the sulfonic acid with aqueous solutions of the metals salts and alkali, alkaline earth metal or ammonium salts. The catalysts have at least 50% of the sulfonic acid groups neutralized with metal ions and the balance of the sulfonic acid groups neutralized with alkali, alkaline earth ions or ammonium ions.

  12. Conductive ceramic composition and method of preparation (United States)

    Smith, J.L.; Kucera, E.H.


    A ceramic anode composition is formed of a multivalent metal oxide or oxygenate such as an alkali metal, transition metal oxygenate. The anode is prepared as a non-stoichiometric crystalline structure by reaction and conditioning in a hydrogen gas cover containing minor proportions of carbon dioxide and water vapor. The structure exhibits a single phase and substantially enhanced electrical conductivity over that of the corresponding stoichiometric structure. Unexpectedly, such oxides and oxygenates are found to be stable in the reducing anode fuel gas of a molten carbonate fuel cell. 4 figures.

  13. High-Throughput Parallel Proteomic Sample Preparation Using 96-Well Polyvinylidene Fluoride (PVDF) Membranes and C18 Purification Plates. (United States)

    Bennike, Tue Bjerg; Steen, Hanno


    Meaningful proteomic-based biomarker discovery projects using primary human-derived specimens require the analysis of hundreds of samples in order to address the issue of interpersonal variability. Thus, robust high-throughput methods for the digestion of plasma samples are a prerequisite for such large clinical proteomic studies with hundreds of samples. Commonly used sample preparation methods are often difficult to parallelize and/or automate. Herein we describe a method for parallel 96-well plate-based sample preparation. Protein digestion is performed in 96-well polyvinylidene fluoride (PVDF) membrane plates and the subsequent purification in 96-well reversed phase C18 purification plates, enabling the usage of multichannel pipettes in all steps. The protocol can be applied using neat or depleted plasma/serum samples, but has also proven effective with other sample types.

  14. Methods of preparation of Swazi traditional fermented foods


    Simatende, Protus; Gadaga, Tendekayi Henry; Jabulani Nkambule, Stanley; SIWELA,Muthulisi


    Background: Fermentation is an age old technique of preserving food in many communities. A wide range of fermented products are prepared by varying the types of raw materials, utensils, and fermentation times. Several fermented foods are consumed in Swaziland. A survey of the types of fermented foods, preparation methods, and utensils used was done in the Hhohho region of Swaziland. The current study aimed at documenting the preparation methods of emahewu, emasi, umcombotsi, and buganu at hou...

  15. Novel Method Of Preparing Vaccines | NCI Technology Transfer Center | TTC (United States)

    This invention from the NCI Cancer and Inflammation Program describes methods to prepare vaccines for the treatment of human immunodeficiency virus (HIV) infections. The National Cancer Institute's Cancer and Inflammation Program seeks parties interested in licensing or collaborative research to further develop, evaluate, or commercialize novel methods of preparing vaccines.

  16. Method for preparing small volume reaction containers (United States)

    Retterer, Scott T.; Doktycz, Mitchel J.


    Engineered reaction containers that can be physically and chemically defined to control the flux of molecules of different sizes and charge are disclosed. Methods for constructing small volume reaction containers through a combination of etching and deposition are also disclosed. The methods allow for the fabrication of multiple devices that possess features on multiple length scales, specifically small volume containers with controlled porosity on the nanoscale.

  17. Methods to Prepare Aluminum Salt-Adjuvanted Vaccines. (United States)

    Thakkar, Sachin G; Cui, Zhengrong


    Many human vaccines contain certain insoluble aluminum salts such as aluminum oxyhydroxide and aluminum hydroxyphosphate as vaccine adjuvants to boost the immunogenicity of the vaccines. Aluminum salts have been used as vaccine adjuvants for decades and have an established, favorable safety profile. However, preparing aluminum salts and aluminum salt-adjuvanted vaccines in a consistent manner remains challenging. This chapter discusses methods to prepare aluminum salts and aluminum salt-adjuvanted vaccines, factors to consider during preparation, and methods to characterize the vaccines after preparation.


    Energy Technology Data Exchange (ETDEWEB)

    Edwards, T.; Hera, K.; Coleman, C.


    Evaluation of Defense Waste Processing Facility (DWPF) Chemical Process Cell (CPC) cycle time identified several opportunities to improve the CPC processing time. The Mechanical Systems & Custom Equipment Development (MS&CED) Section of the Savannah River National Laboratory (SRNL) recently completed the evaluation of one of these opportunities - the possibility of using an Isolok sampling valve as an alternative to the Hydragard valve for taking DWPF process samples at the Slurry Mix Evaporator (SME). The use of an Isolok for SME sampling has the potential to improve operability, reduce maintenance time, and decrease CPC cycle time. The SME acceptability testing for the Isolok was requested in Task Technical Request (TTR) HLW-DWPF-TTR-2010-0036 and was conducted as outlined in Task Technical and Quality Assurance Plan (TTQAP) SRNLRP-2011-00145. RW-0333P QA requirements applied to the task, and the results from the investigation were documented in SRNL-STI-2011-00693. Measurement of the chemical composition of study samples was a critical component of the SME acceptability testing of the Isolok. A sampling and analytical plan supported the investigation with the analytical plan directing that the study samples be prepared by a cesium carbonate (Cs{sub 2}CO{sub 3}) fusion dissolution method and analyzed by Inductively Coupled Plasma - Optical Emission Spectroscopy (ICP-OES). The use of the cesium carbonate preparation method for the Isolok testing provided an opportunity for an additional assessment of this dissolution method, which is being investigated as a potential replacement for the two methods (i.e., sodium peroxide fusion and mixed acid dissolution) that have been used at the DWPF for the analysis of SME samples. Earlier testing of the Cs{sub 2}CO{sub 3} method yielded promising results which led to a TTR from Savannah River Remediation, LLC (SRR) to SRNL for additional support and an associated TTQAP to direct the SRNL efforts. A technical report resulting

  19. Insect chromosomes preparing methods for genetic researches

    African Journals Online (AJOL)



    Jan 5, 2009 ... Importantly, each chromosome displays a unique banding pattern. Different staining methods showed the banding pattern (g band), ... with 100 t/mn in a clinical centrifugal machine, with sodium citrate for 1 - 10 min, fixed and dried as described above. Measurement of length are taken on photographs.

  20. Comparison between Properties of Polyurethane Nanocomposites Prepared by Two Different Methods

    Directory of Open Access Journals (Sweden)

    M. Barmar


    Full Text Available In this work, a thermoplastic polyurethane elastomer (TPUE model based on polytetramethylene glycol, toluene diisocyanate and 1,4-butanediol was selected and synthesized. According to this model two types of polyurethane nanocomposites were prepared by in situ polymerization and melt intercalation procedures. The organo-modified nanoclay was used in nanocomposites samples in 0.4 weight percent level. The prepared nanocomposites were studied by WAXD, tensile and thermal analysis. Thermal properties of the nanocomposites were higher than those of pure polyurethane elastomers. Nanocomposites prepared via melt intercalation method showed a lower tensile strength and hardness than those prepared through in situ polymerization method..

  1. System and Method for Isolation of Samples (United States)

    Zhang, Ye (Inventor); Wu, Honglu (Inventor)


    Systems and methods for isolating samples are provided. The system comprises a first membrane and a second membrane disposed within an enclosure. First and second reservoirs can also be disposed within the enclosure and adapted to contain one or more reagents therein. A first valve can be disposed within the enclosure and in fluid communication with the first reservoir, the second reservoir, or both. The first valve can also be in fluid communication with the first or second membranes or both. The first valve can be adapted to selectively regulate the flow of the reagents from the first reservoir, through at least one of the first and second membranes, and into the second reservoir.

  2. High-resolution X-ray diffraction with no sample preparation. (United States)

    Hansford, G M; Turner, S M R; Degryse, P; Shortland, A J


    It is shown that energy-dispersive X-ray diffraction (EDXRD) implemented in a back-reflection geometry is extremely insensitive to sample morphology and positioning even in a high-resolution configuration. This technique allows high-quality X-ray diffraction analysis of samples that have not been prepared and is therefore completely non-destructive. The experimental technique was implemented on beamline B18 at the Diamond Light Source synchrotron in Oxfordshire, UK. The majority of the experiments in this study were performed with pre-characterized geological materials in order to elucidate the characteristics of this novel technique and to develop the analysis methods. Results are presented that demonstrate phase identification, the derivation of precise unit-cell parameters and extraction of microstructural information on unprepared rock samples and other sample types. A particular highlight was the identification of a specific polytype of a muscovite in an unprepared mica schist sample, avoiding the time-consuming and difficult preparation steps normally required to make this type of identification. The technique was also demonstrated in application to a small number of fossil and archaeological samples. Back-reflection EDXRD implemented in a high-resolution configuration shows great potential in the crystallographic analysis of cultural heritage artefacts for the purposes of scientific research such as provenancing, as well as contributing to the formulation of conservation strategies. Possibilities for moving the technique from the synchrotron into museums are discussed. The avoidance of the need to extract samples from high-value and rare objects is a highly significant advantage, applicable also in other potential research areas such as palaeontology, and the study of meteorites and planetary materials brought to Earth by sample-return missions.

  3. Firstborn microcrystallization method to prepare nanocapsules containing artesunate


    Xin-Cai Xiao; Zong-Guo Hong


    Xin-Cai Xiao, Zong-Guo HongSchool of Pharmacy, South-Central University for Nationalities, Wuhan, 430074, ChinaAbstract: We developed a novel method to prepare nanocapsules. A solute often crystallizes when its solubility alters from one solvent to another, and its firstborn crystals are used as templates to prepare nanocapsules for the first time, which is called firstborn microcrystallization method. By using this method, the maximum diameter of the nanocapsules including artesunate is abou...

  4. Microfluidic desalination : capacitive deionization on chip for microfluidic sample preparation

    NARCIS (Netherlands)

    Roelofs, Susan Helena


    The main aim of the work described in this thesis is to implement the desalination technique capacitive deionization (CDI) on a microfluidic chip to improve the reproducibility in the analysis of biological samples for drug development. Secondly, microfluidic CDI allows for the in situ study of ion

  5. Sample Preparation (SS): SE60_SS01 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available and a zirconia bead for 6 min at 20 Hz. Samples were centrifuged at 15 000 g for 10 min. ..., using a mixer mill (MM 300, Retsch, Haan, Germany, http://www.rets

  6. Sample Preparation (SS): SE55_SS01 [Metabolonote[Archive

    Lifescience Database Archive (English)

    Full Text Available ,000g for 10 min and filtration (Ultrafree-MC filter, 0.2 mm, Millipore), the sample extracts were applied t...o an HLB mElution plate (Waters) equilibrated with 80% aqueous methanol containing 0.1% acetic acid. ...

  7. Sample preparation of Medicago sativa L. hay for chemical analysis ...

    African Journals Online (AJOL)

    The objective of this study was to quantify the effect of the grinding procedure on the moisture and crude protein concentration of a ground Medicago sativa L. hay sample for quality grading. An additional aim was to investigate the accuracy of electronic moisture testers (EMT). Variance of analyses revealed significant ...

  8. Nanoemulsions: the properties, methods of preparation and promising applications (United States)

    Koroleva, M. Yu; Yurtov, Evgenii V.


    The properties of nanoemulsions and various methods for their preparation including the high-energy and low-energy emulsification methods and the combined methods are reviewed. Among the high-energy methods, the emphasis is placed on high-energy stirring, ultrasonic emulsification, high-pressure homogenization including microfluidics and membrane emulsification. Among the low-energy emulsification methods, the attention is focused on the phase inversion temperature method, the emulsion inversion point method and the spontaneous emulsification. Using a combined method, which includes the high-energy and low-energy emulsification, it is possible to prepare reverse nanoemulsions in highly viscous systems. Main advantages and limitations of different methods of nanoemulsion preparation are discussed and the potential fields of nanoemulsion applications are considered. The bibliography includes 255 references.

  9. Sample Preparation for Headspace GC Analysis of Residual Solvents in Hyaluronic Acid Derivative Fiber

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hoon Joo; Kim, Dong Min; Yang, Jeong Soo [LG life Sciences, Ltd./R and D Park, Daejeon (Korea, Republic of); Kim, Chan Wha [Korea University, Seoul (Korea, Republic of)


    The aim of this study is to develop efficient sample preparation method for HS-GC analysis of residual solvents in HA derivative fiber. Compared to direct extraction of residual solvents from HA derivative fiber, the extraction through the hydrolysis of HA derivative fiber by HAse gave more complete and higher reproducible quantification of residual solvent. To validate HS-GC analysis method of residual solvents, specificity, limits of detection and quantification, linearity, accuracy and precision are investigated in the study. HA derivative fiber was hydrolyzed using HAse for headspace gas chromatographic analysis of residual solvents of ethanol, acetone and isopropanol in HA derivative fiber. This study showed that the developed method had specificity, linearity, accuracy and precision. In addition, it demonstrated that HS-GC coupled with matrix-breaking method such as hydrolysis was available for the determination of residual solvents in a matrix like HA derivative fiber.

  10. Transuranic waste characterization sampling and analysis methods manual. Revision 1

    Energy Technology Data Exchange (ETDEWEB)

    Suermann, J.F.


    This Methods Manual provides a unified source of information on the sampling and analytical techniques that enable Department of Energy (DOE) facilities to comply with the requirements established in the current revision of the Transuranic Waste Characterization Quality Assurance Program Plan (QAPP) for the Waste Isolation Pilot Plant (WIPP) Transuranic (TRU) Waste Characterization Program (the Program) and the WIPP Waste Analysis Plan. This Methods Manual includes all of the testing, sampling, and analytical methodologies accepted by DOE for use in implementing the Program requirements specified in the QAPP and the WIPP Waste Analysis Plan. The procedures in this Methods Manual are comprehensive and detailed and are designed to provide the necessary guidance for the preparation of site-specific procedures. With some analytical methods, such as Gas Chromatography/Mass Spectrometry, the Methods Manual procedures may be used directly. With other methods, such as nondestructive characterization, the Methods Manual provides guidance rather than a step-by-step procedure. Sites must meet all of the specified quality control requirements of the applicable procedure. Each DOE site must document the details of the procedures it will use and demonstrate the efficacy of such procedures to the Manager, National TRU Program Waste Characterization, during Waste Characterization and Certification audits.

  11. Rapid microbial sample preparation from blood using a novel concentration device.

    Directory of Open Access Journals (Sweden)

    Anna K Boardman

    Full Text Available Appropriate care for bacteremic patients is dictated by the amount of time needed for an accurate diagnosis. However, the concentration of microbes in the blood is extremely low in these patients (1-100 CFU/mL, traditionally requiring growth (blood culture or amplification (e.g., PCR for detection. Current culture-based methods can take a minimum of two days, while faster methods like PCR require a sample free of inhibitors (i.e., blood components. Though commercial kits exist for the removal of blood from these samples, they typically capture only DNA, thereby necessitating the use of blood culture for antimicrobial testing. Here, we report a novel, scaled-up sample preparation protocol carried out in a new microbial concentration device. The process can efficiently lyse 10 mL of bacteremic blood while maintaining the microorganisms' viability, giving a 30-μL final output volume. A suite of six microorganisms (Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, Pseudomonas aeruginosa, and Candida albicans at a range of clinically relevant concentrations was tested. All of the microorganisms had recoveries greater than 55% at the highest tested concentration of 100 CFU/mL, with three of them having over 70% recovery. At the lowest tested concentration of 3 CFU/mL, two microorganisms had recoveries of ca. 40-50% while the other four gave recoveries greater than 70%. Using a Taqman assay for methicillin-sensitive S. aureus (MSSAto prove the feasibility of downstream analysis, we show that our microbial pellets are clean enough for PCR amplification. PCR testing of 56 spiked-positive and negative samples gave a specificity of 0.97 and a sensitivity of 0.96, showing that our sample preparation protocol holds great promise for the rapid diagnosis of bacteremia directly from a primary sample.

  12. Automated dried blood spots standard and QC sample preparation using a robotic liquid handler. (United States)

    Yuan, Long; Zhang, Duxi; Aubry, Anne-Francoise; Arnold, Mark E


    A dried blood spot (DBS) bioanalysis assay involves many steps, such as the preparation of standard (STD) and QC samples in blood, the spotting onto DBS cards, and the cutting-out of the spots. These steps are labor intensive and time consuming if done manually, which, therefore, makes automation very desirable in DBS bioanalysis. A robotic liquid handler was successfully applied to the preparation of STD and QC samples in blood and to spot the blood samples onto DBS cards using buspirone as the model compound. This automated preparation was demonstrated to be accurate and consistent. However the accuracy and precision of automated preparation were similar to those from manual preparation. The effect of spotting volume on accuracy was evaluated and a trend of increasing concentrations of buspirone with increasing spotting volumes was observed. The automated STD and QC sample preparation process significantly improved the efficiency, robustness and safety of DBS bioanalysis.

  13. Enzymatic tissue digestion as an alternative sample preparation approach for quantitative analysis using liquid chromatography-tandem mass spectrometry. (United States)

    Yu, Chongwoo; Penn, Lara D; Hollembaek, John; Li, Wenlin; Cohen, Lucinda H


    Compound extraction from biological tissue often presents a challenge for the bioanalytical chemist. Labor-intensive homogenization or sonication of whole or powdered tissue is performed before compounds can be extracted and analyzed. Enzymatic digestion is commonly used for tissue dissociation and cell harvesting and offers the advantages of unattended sample preparation, potential automation, and low cost. The feasibility of enzymatic digestion as an alternate tissue preparation technique was evaluated for bioanalysis of drugs in conjunction with LC/MS/MS. Two different enzymes (collagenase and proteinase K) that are known to degrade connective tissues to allow tissue dissolution were chosen for evaluation, employing well-known antidepressants desipramine and fluoxetine as test compounds in dog and rat brain tissue. Comparison between enzymatic digestion and conventional homogenization tissue preparation was performed, including investigation of matrix ionization suppression of both methods using a postcolumn infusion system. Results showed that enzymatic digestion has extraction efficiency comparable to homogenization. Matrix ionization suppression was not observed for either the test compounds evaluated or the sample extraction method. Test compound levels of incurred tissue samples prepared by enzymatic digestion were in good agreement with the values obtained by the conventional homogenization tissue preparation, indicating that enzymatic digestion is an appropriate tissue sample preparation method.

  14. Recent advances in metal-organic frameworks and covalent organic frameworks for sample preparation and chromatographic analysis. (United States)

    Wang, Xuan; Ye, Nengsheng


    In the field of analytical chemistry, sample preparation and chromatographic separation are two core procedures. The means by which to improve the sensitivity, selectivity and detection limit of a method have become a topic of great interest. Recently, porous organic frameworks, such as metal-organic frameworks (MOFs) and covalent organic frameworks (COFs), have been widely used in this research area because of their special features, and different methods have been developed. This review summarizes the applications of MOFs and COFs in sample preparation and chromatographic stationary phases. The MOF- or COF-based solid-phase extraction (SPE), solid-phase microextraction (SPME), gas chromatography (GC), high-performance liquid chromatography (HPLC) and capillary electrochromatography (CEC) methods are described. The excellent properties of MOFs and COFs have resulted in intense interest in exploring their performance and mechanisms for sample preparation and chromatographic separation. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Sample preparation for precise and quantitative electron holographic analysis of semiconductor devices. (United States)

    Han, Myung-Geun; Li, Jing; Xie, Qianghua; Fejes, Peter; Conner, James; Taylor, Bill; McCartney, Martha R


    Wedge polishing was used to prepare one-dimensional Si n-p junction and Si p-channel metal-oxide-silicon field effect transistor (pMOSFET) samples for precise and quantitative electrostatic potential analysis using off-axis electron holography. To avoid artifacts associated with ion milling, cloth polishing with 0.02-microm colloidal silica suspension was used for final thinning. Uniform thickness and no significant charging were observed by electron holography analysis for samples prepared entirely by this method. The effect of sample thickness was investigated and the minimum thickness for reliable results was found to be approximately 160 nm. Below this thickness, measured phase changes were smaller than expected. For the pMOSFET sample, quantitative analysis of two-dimensional electrostatic potential distribution showed that the metallurgical gate length (separation between two extension junctions) was approximately 54 nm, whereas the actual gate length was measured to be approximately 70 nm by conventional transmission electron microscopy. Thus, source and drain junction encroachment under the gate was 16 nm.

  16. Effect of method of preparation on hydrodesulphurization activity of ...

    Indian Academy of Sciences (India)

    molybdenum sulphide catalysts through precipitation from homogeneous solution (PFHS) technique with the. Mo content varying from 2–12 wt%. The prepared catalysts were evaluated for thiophene hydrodesulphuriza- tion catalytic activities at 400. ◦. C. Catalysts prepared through PFHS method resulted in highly dispersed ...

  17. New methods for preparing mercury-based ferrofluids

    DEFF Research Database (Denmark)

    Linderoth, Søren; Rasmussen, L.H.; Mørup, Steen


    Metallic ferrofluids based on magnetic particles in mercury have been produced by two new methods. Alloy particles of Fe-B, Fe-Co-B, Fe-Ni-B, and Co-B were prepared by reduction of the transition metal ions in aqueous solutions by NaBH4 and subsequently suspended in mercury. In one preparation...

  18. Formulation and method for preparing gels comprising hydrous hafnium oxide (United States)

    Collins, Jack L; Hunt, Rodney D; Montgomery, Frederick C


    Formulations useful for preparing hydrous hafnium oxide gels contain a metal salt including hafnium, an acid, an organic base, and a complexing agent. Methods for preparing gels containing hydrous hafnium oxide include heating a formulation to a temperature sufficient to induce gel formation, where the formulation contains a metal salt including hafnium, an acid, an organic base, and a complexing agent.

  19. Formulation and method for preparing gels comprising hydrous cerium oxide (United States)

    Collins, Jack L; Chi, Anthony


    Formulations useful for preparing hydrous cerium oxide gels contain a metal salt including cerium, an organic base, and a complexing agent. Methods for preparing gels containing hydrous cerium oxide include heating a formulation to a temperature sufficient to induce gel formation, where the formulation contains a metal salt including cerium, an organic base, and a complexing agent.

  20. Formulation and method for preparing gels comprising hydrous aluminum oxide (United States)

    Collins, Jack L.


    Formulations useful for preparing hydrous aluminum oxide gels contain a metal salt including aluminum, an organic base, and a complexing agent. Methods for preparing gels containing hydrous aluminum oxide include heating a formulation to a temperature sufficient to induce gel formation, where the formulation contains a metal salt including aluminum, an organic base, and a complexing agent.

  1. Antithrombogenic and antibiotic composition and methods of preparation thereof (United States)

    Hermes, Robert E.


    Antithrombogenic and antibiotic composition of matter and method of preparation thereof. A random copolymer of a component of garlic and a biocompatible polymer has been prepared and found to exhibit antithrombogenic and antibiotic properties. Polymerization occurs selectively at the vinyl moiety in 2-vinyl-4H-1,3-dithiin when copolymerized with N-vinyl pyrrolidone.

  2. How to prepare cytological samples for molecular testing. (United States)

    Bellevicine, Claudio; Malapelle, Umberto; Vigliar, Elena; Pisapia, Pasquale; Vita, Giulia; Troncone, Giancarlo


    This review is focused on the challenges in standardising and optimising molecular testing workflow in cytopathology. Although cytological samples yield optimal quality DNA, whose minimal amounts in most cases suffice even for multigene mutational profiling, the success of molecular testing is strongly dependent on standardised preanalytical protocols for maximising DNA yield and quality. Sample cytopreparation influences, even more, the quality of RNA and consequently the potential success of reverse transcription-PCR. Here, the educational and technical involvement of the cytopathologist as a relevant component of a multidisciplinary team, in the issues related to test request, specimen collection, fixation, processing, staining, tumour fraction enrichment, DNA quality/quantity assessment and storage conditions is discussed. In addition, the specific sample requirements related to more recent technological developments are examined, underlining the modern role of the cytopathologist, whose continuous education is crucial to meet the opportunities of molecular medicine. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  3. The effect of sample preparation and testing procedure on the ...

    African Journals Online (AJOL)

    It is concluded that the plasticity chart should be regarded as a guidance tool only for evaluating the likely properties of laterite soils. It should not be used to classify them based on conventional methods without due deference to their genesis. Unlike soils for which the Casagrande Plastcity Chart, as adopted in the United ...


    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, S.; Culligan, B.; Noyes, G.


    A new rapid method for the determination of actinides in soil and sediment samples has been developed at the Savannah River Site Environmental Lab (Aiken, SC, USA) that can be used for samples up to 2 grams in emergency response situations. The actinides in soil method utilizes a rapid sodium hydroxide fusion method, a lanthanum fluoride soil matrix removal step, and a streamlined column separation process with stacked TEVA, TRU and DGA Resin cartridges. Lanthanum was separated rapidly and effectively from Am and Cm on DGA Resin. Vacuum box technology and rapid flow rates are used to reduce analytical time. Alpha sources are prepared using cerium fluoride microprecipitation for counting by alpha spectrometry. The method showed high chemical recoveries and effective removal of interferences. This new procedure was applied to emergency soil samples received in the NRIP Emergency Response exercise administered by the National Institute for Standards and Technology (NIST) in April, 2009. The actinides in soil results were reported within 4-5 hours with excellent quality.

  5. Can matrix solid phase dispersion (MSPD) be more simplified? Application of solventless MSPD sample preparation method for GC-MS and GC-FID analysis of plant essential oil components. (United States)

    Wianowska, Dorota; Dawidowicz, Andrzej L


    This paper proposes and shows the analytical capabilities of a new variant of matrix solid phase dispersion (MSPD) with the solventless blending step in the chromatographic analysis of plant volatiles. The obtained results prove that the use of a solvent is redundant as the sorption ability of the octadecyl brush is sufficient for quantitative retention of volatiles from 9 plants differing in their essential oil composition. The extraction efficiency of the proposed simplified MSPD method is equivalent to the efficiency of the commonly applied variant of MSPD with the organic dispersing liquid and pressurized liquid extraction, which is a much more complex, technically advanced and highly efficient technique of plant extraction. The equivalency of these methods is confirmed by the variance analysis. The proposed solventless MSPD method is precise, accurate, and reproducible. The recovery of essential oil components estimated by the MSPD method exceeds 98%, which is satisfactory for analytical purposes. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. RNA sample preparation applied to gene expression profiling for the horse biological passport. (United States)

    Bailly-Chouriberry, Ludovic; Baudoin, Florent; Cormant, Florence; Glavieux, Yohan; Loup, Benoit; Garcia, Patrice; Popot, Marie-Agnès; Bonnaire, Yves


    The improvement of doping control is an ongoing race. Techniques to fight doping are usually based on the direct detection of drugs or their metabolites by analytical methods such as chromatography hyphenated to mass spectrometry after ad hoc sample preparation. Nowadays, omic methods constitute an attractive development and advances have been achieved particularly by application of molecular biology tools for detection of anabolic androgenic steroids (AAS), erythropoiesis-stimulating agent (ESA), or to control human growth hormone misuses. These interesting results across different animal species have suggested that modification of gene expression offers promising new methods of improving the window of detection of banned substances by targeting their effects on blood cell gene expression. In this context, the present study describes the possibility of using a modified version of the dedicated Human IVD (in vitro Diagnostics) PAXgene® Blood RNA Kit for horse gene expression analysis in blood collected on PAXgene® tubes applied to the horse biological passport. The commercial kit was only approved for human blood samples and has required an optimization of specific technical requirements for equine blood samples. Improvements and recommendations were achieved for sample collection, storage and RNA extraction procedure. Following these developments, RNA yield and quality were demonstrated to be suitable for downstream gene expression analysis by qPCR techniques. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

  7. Preparation of Magnetic Hollow Molecularly Imprinted Polymers for Detection of Triazines in Food Samples. (United States)

    Wang, Aixiang; Lu, Hongzhi; Xu, Shoufang


    Novel magnetic hollow molecularly imprinted polymers (M-H-MIPs) were proposed for highly selective recognition and fast enrichment of triazines in food samples. M-H-MIPs were prepared on the basis of multi-step swelling polymerization, followed by in situ growth of magnetic Fe3O4 nanoparticles on the surface of hollow molecularly imprinted polymers (H-MIPs). Transmission electron microscopy and scanning electron microscopy confirmed the successful immobilization of Fe3O4 nanoparticles on the surface of H-MIPs. M-H-MIPs could be separated simply using an external magnet. The binding adsorption results indicated that M-H-MIPs displayed high binding capacity and fast mass transfer property and class selective property for triazines. Langmuir isotherm and pseudo-second-order kinetic models fitted the best adsorption models for M-H-MIPs. M-H-MIPs were used to analyze atrazine, simazine, propazine, and terbuthylazine in corn, wheat, and soybean samples. Satisfactory recoveries were in the range of 80.62-101.69%, and relative standard deviation was lower than 5.2%. Limits of detection from 0.16 to 0.39 μg L(-1) were obtained. When the method was applied to test positive samples that were contaminated with triazines, the results agree well with those obtained from an accredited method. Thus, the M-H-MIP-based dispersive solid-phase extraction method proved to be a convenient and practical platform for detection of triazines in food samples.

  8. Methods and apparatuses for preparing upgraded pyrolysis oil (United States)

    Brandvold, Timothy A; Baird, Lance Awender; Frey, Stanley Joseph


    Methods and apparatuses for preparing upgraded pyrolysis oil are provided herein. In an embodiment, a method of preparing upgraded pyrolysis oil includes providing a biomass-derived pyrolysis oil stream having an original oxygen content. The biomass-derived pyrolysis oil stream is hydrodeoxygenated under catalysis in the presence of hydrogen to form a hydrodeoxygenated pyrolysis oil stream comprising a cyclic paraffin component. At least a portion of the hydrodeoxygenated pyrolysis oil stream is dehydrogenated under catalysis to form the upgraded pyrolysis oil.

  9. A new method of preparing single-walled carbon nanotubes

    Indian Academy of Sciences (India)

    A novel method of purification for single-walled carbon nanotubes, prepared by an arc-discharge method, is described. The method involves a combination of acid washing followed by high temperature hydrogen treatment to remove the metal nanoparticles and amorphous carbon present in the as-synthesized singlewalled ...

  10. The preparation method of terahertz monolithic integrated device (United States)

    Zhang, Cong; Su, Bo; He, Jingsuo; Zhang, Hongfei; Wu, Yaxiong; Zhang, Shengbo; Zhang, Cunlin


    The terahertz monolithic integrated device is to integrate the pumping area of the terahertz generation, the detection area of the terahertz receiving and the metal waveguide of terahertz transmission on the same substrate. The terahertz generation and detection device use a photoconductive antenna structure the metal waveguide use a microstrip line structure. The evanescent terahertz-bandwidth electric field extending above the terahertz transmission line interacts with, and is modified by, overlaid dielectric samples, thus enabling the characteristic vibrational absorption resonances in the sample to be probed. In this device structure, since the semiconductor substrate of the photoconductive antenna is located between the strip conductor and the dielectric layer of the microstrip line, and the semiconductor substrate cannot grow on the dielectric layer directly. So how to prepare the semiconductor substrate of the photoconductive antenna and how to bond the semiconductor substrate to the dielectric layer of the microstrip line is a key step in the terahertz monolithic integrated device. In order to solve this critical problem, the epitaxial wafer structure of the two semiconductor substrates is given and transferred to the desired substrate by two methods, respectively.

  11. /dopamine films prepared by sol-gel method (United States)

    Valverde-Aguilar, G.; Prado-Prone, G.; Vergara-Aragón, P.; Garcia-Macedo, J.; Santiago, Patricia; Rendón, Luis


    Dopamine was encapsulated into nanoporous amorphous TiO2 matrix by sol-gel method under atmospheric conditions. A second sample was obtained by the addition of the crown-ether 15C5 in this previous sample. Thin films were spin-coated on glass wafers. No heat treatment was employed in both films. All films were characterized using infrared spectroscopy, high resolution transmission electronic microscopy, X-ray diffraction, optical absorption and scanning electronic microscopy. Despite the films prepared with 15C5 were no calcined, a partial crystallization was identified. Anatase and rutile nanoparticles with sizes of 4-5 nm were obtained. Photoconductivity technique was used to determine the charge transport mechanism on these films. Experimental data were fitted with straight lines at darkness and under illumination wavelengths at 320, 400, and 515 nm. It indicates an ohmic behavior. Photovoltaic and photoconductivity parameters were determined from the current density vs. the applied-electrical-field results. Amorphous film has bigger photovoltaic and photoconductive parameters than the partially crystalline film. Results observed in the present investigation prove that the nanoporous TiO2 matrix can protect the dopamine inhibiting its chemical instability. This fact modifies the optical, physical and electrical properties of the film, and is intensified when 15C5 is added.

  12. Optical Methods for Identifying Hard Clay Core Samples During Petrophysical Studies (United States)

    Morev, A. V.; Solovyeva, A. V.; Morev, V. A.


    X-ray phase analysis of the general mineralogical composition of core samples from one of the West Siberian fields was performed. Electronic absorption spectra of the clay core samples with an added indicator were studied. The speed and availability of applying the two methods in petrophysical laboratories during sample preparation for standard and special studies were estimated.

  13. Quality Assessment of Platelet-Rich Fibrin-Like Matrix Prepared from Whole Blood Samples after Extended Storage. (United States)

    Kawabata, Hideo; Isobe, Kazushige; Watanabe, Taisuke; Okudera, Toshimitsu; Nakamura, Masayuki; Suzuki, Masashi; Ryu, Jietsu; Kitamura, Yutaka; Okudera, Hajime; Okuda, Kazuhiro; Nakata, Koh; Kawase, Tomoyuki


    The platelet-rich fibrin-like matrix (PRFM) is usually prepared onsite and immediately used for regenerative therapy. Nonetheless, to meet the clinical necessity of preserving the PRFM without quality deterioration, we developed a method for preparation of PRFMs from short-term-stored whole blood (WB) samples. In this study, to evaluate the practical expiration date of storage, we extended the storage time of WB samples from 2 to 7 days and assessed the quality of the resulting PRFMs. WB samples collected with acid-citrate-dextrose were stored with gentle agitation at ambient temperature. To prepare PRFMs, the stored WB samples were mixed with CaCl₂ in glass tubes and centrifuged. Fibrin fiber networks, CD41 and CD62P expression, and Platelet Derived Growth Factor-BB (PDGF-BB) levels were examined by scanning electron microscopy (SEM), flow cytometry, and an Enzyme-Linked ImmunoSorbent Assay (ELISA), respectively. Long-term storage had no significant effect on either blood cell counts or platelet functions tested. The resulting PRFMs were visually identical to freshly prepared ones. PDGF-BB levels did not markedly decrease in a time-dependent manner. However, fibrin fibers gradually became thinner after storage. Although the coagulation activity may diminish, we propose that PRFMs can be prepared-without evident loss of quality-from WB samples stored for up to 7 days by our previously developed method.

  14. Study of interaction between croscarmellose and escitalopram during sample preparation. (United States)

    Larsen, Jesper; Melander, Claes


    During routine analysis of an escitalopram tablet formulation, it was seen that there was a systematic deviation between content uniformity (CU - one tablet analysis) and assay analysis (ten pooled tablets). In the presence of the excipients from the tablet, it was found that the extraction of the active pharmaceutical ingredient (API) was incomplete. It was shown that the commonly used tablet disintegrant croscarmellose sodium (crosslinked carboxy-methyl cellulose) had a significant interaction with escitalopram. This was later found to be the explanation for the lower extraction during assay testing. Under normal conditions, the extraction took place in acidic medium which caused protonation of the amine and thereby the interaction of charged species in solution. The interaction of API was studied further with pure croscarmellose and the entire tablet matrix. A range of conditions was considered, including altering extraction volumes, organic solvents, pH of the extraction solvent, and addition of competitive binder in various concentrations. It was seen that arginine was the most effective cationic competitive binder of those tested and that adding it at a suitable concentration level could significantly improve the analytical methods. In the present case, an improvement in recovery was from 98.5% to almost 100% was achieved.

  15. Technical Evaluation of Sample-Processing, Collection, and Preservation Methods (United States)


    enhanced situational awareness of biological threats to the environment, human health, agriculture , and food supplies. Specifically mentioned is the...preparing for the possibility of biologically based attacks on military, civilian, or agricultural targets. To be fully prepared for this...from the various collected samples was extracted using an identical process—the Blood and Tissue Midi Preparation Kit (Qiagen, Inc.; Valencia , CA)—and

  16. Intelligent front-end sample preparation tool using acoustic streaming.

    Energy Technology Data Exchange (ETDEWEB)

    Cooley, Erika J.; McClain, Jaime L.; Murton, Jaclyn K.; Edwards, Thayne L.; Achyuthan, Komandoor E.; Branch, Darren W.; Clem, Paul Gilbert; Anderson, John Mueller; James, Conrad D.; Smith, Gennifer; Kotulski, Joseph Daniel


    We have successfully developed a nucleic acid extraction system based on a microacoustic lysis array coupled to an integrated nucleic acid extraction system all on a single cartridge. The microacoustic lysing array is based on 36{sup o} Y cut lithium niobate, which couples bulk acoustic waves (BAW) into the microchannels. The microchannels were fabricated using Mylar laminates and fused silica to form acoustic-fluidic interface cartridges. The transducer array consists of four active elements directed for cell lysis and one optional BAW element for mixing on the cartridge. The lysis system was modeled using one dimensional (1D) transmission line and two dimensional (2D) FEM models. For input powers required to lyse cells, the flow rate dictated the temperature change across the lysing region. From the computational models, a flow rate of 10 {micro}L/min produced a temperature rise of 23.2 C and only 6.7 C when flowing at 60 {micro}L/min. The measured temperature changes were 5 C less than the model. The computational models also permitted optimization of the acoustic coupling to the microchannel region and revealed the potential impact of thermal effects if not controlled. Using E. coli, we achieved a lysing efficacy of 49.9 {+-} 29.92 % based on a cell viability assay with a 757.2 % increase in ATP release within 20 seconds of acoustic exposure. A bench-top lysing system required 15-20 minutes operating up to 58 Watts to achieve the same level of cell lysis. We demonstrate that active mixing on the cartridge was critical to maximize binding and release of nucleic acid to the magnetic beads. Using a sol-gel silica bead matrix filled microchannel the extraction efficacy was 40%. The cartridge based magnetic bead system had an extraction efficiency of 19.2%. For an electric field based method that used Nafion films, a nucleic acid extraction efficiency of 66.3 % was achieved at 6 volts DC. For the flow rates we tested (10-50 {micro}L/min), the nucleic acid extraction

  17. Review of sample preparation strategies for MS-based metabolomic studies in industrial biotechnology. (United States)

    Causon, Tim J; Hann, Stephan


    Fermentation and cell culture biotechnology in the form of so-called "cell factories" now play an increasingly significant role in production of both large (e.g. proteins, biopharmaceuticals) and small organic molecules for a wide variety of applications. However, associated metabolic engineering optimisation processes relying on genetic modification of organisms used in cell factories, or alteration of production conditions remain a challenging undertaking for improving the final yield and quality of cell factory products. In addition to genomic, transcriptomic and proteomic workflows, analytical metabolomics continues to play a critical role in studying detailed aspects of critical pathways (e.g. via targeted quantification of metabolites), identification of biosynthetic intermediates, and also for phenotype differentiation and the elucidation of previously unknown pathways (e.g. via non-targeted strategies). However, the diversity of primary and secondary metabolites and the broad concentration ranges encompassed during typical biotechnological processes means that simultaneous extraction and robust analytical determination of all parts of interest of the metabolome is effectively impossible. As the integration of metabolome data with transcriptome and proteome data is an essential goal of both targeted and non-targeted methods addressing production optimisation goals, additional sample preparation steps beyond necessary sampling, quenching and extraction protocols including clean-up, analyte enrichment, and derivatisation are important considerations for some classes of metabolites, especially those present in low concentrations or exhibiting poor stability. This contribution critically assesses the potential of current sample preparation strategies applied in metabolomic studies of industrially-relevant cell factory organisms using mass spectrometry-based platforms primarily coupled to liquid-phase sample introduction (i.e. flow injection, liquid

  18. A novel method for the preparation of electrophoretic display microcapsules

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Xiao-Meng; He, Jing; Liu, Sheng-Yun [State Key Laboratory of Organic-Inorganic Composites, Beijing University of Chemical Technology, Beijing 100029 (China); Chen, Jian-Feng [State Key Laboratory of Organic-Inorganic Composites, Beijing University of Chemical Technology, Beijing 100029 (China); Research Center of the Ministry of Education for High Gravity Engineering and Technology, Beijing University of Chemical Technology, Beijing 100029 (China); Le, Yuan, E-mail: [State Key Laboratory of Organic-Inorganic Composites, Beijing University of Chemical Technology, Beijing 100029 (China)


    Highlights: • The electrophoretic display microcapsules were prepared by coaxial jet method aided by gas spray. • The positions of inner tube, liquid and gas flow rate of the process were investigated. • The size and shell thickness of the prepared microcapsules were controllable. • The prepared microcapsules had high coating ratio and exhibit reversible response to DC field. - Abstract: The narrow distributed electrophoretic display microcapsules containing electrophoretic ink were prepared using coaxial jet method aided by gas spray. Experimental results showed the size and shell thickness of the microcapsules could be controlled by adjusting flow rates of core and shell fluids as well as gas. The as-prepared white and red microcapsules, with average size of 100 and 200 μm respectively, had high coating ratio (above 90%) and exhibited reversible response to DC electric field. Compared with the approach of other microencapsulation methods, the new technique not only has a simple procedure but also provides a more effective way of size control. This novel method is expected to prepare microcapsules with potential application in the fields of electronic paper and other material science.

  19. Development of an automated data processing method for sample to sample comparison of seized methamphetamines. (United States)

    Choe, Sanggil; Lee, Jaesin; Choi, Hyeyoung; Park, Yujin; Lee, Heesang; Pyo, Jaesung; Jo, Jiyeong; Park, Yonghoon; Choi, Hwakyung; Kim, Suncheun


    The information about the sources of supply, trafficking routes, distribution patterns and conspiracy links can be obtained from methamphetamine profiling. The precursor and synthetic method for the clandestine manufacture can be estimated from the analysis of minor impurities contained in methamphetamine. Also, the similarity between samples can be evaluated using the peaks that appear in chromatograms. In South Korea, methamphetamine was the most popular drug but the total seized amount of methamphetamine whole through the country was very small. Therefore, it would be more important to find the links between samples than the other uses of methamphetamine profiling. Many Asian countries including Japan and South Korea have been using the method developed by National Research Institute of Police Science of Japan. The method used gas chromatography-flame ionization detector (GC-FID), DB-5 column and four internal standards. It was developed to increase the amount of impurities and minimize the amount of methamphetamine. After GC-FID analysis, the raw data have to be processed. The data processing steps are very complex and require a lot of time and effort. In this study, Microsoft Visual Basic Application (VBA) modules were developed to handle these data processing steps. This module collected the results from the data into an Excel file and then corrected the retention time shift and response deviation generated from the sample preparation and instruments analysis. The developed modules were tested for their performance using 10 samples from 5 different cases. The processed results were analyzed with Pearson correlation coefficient for similarity assessment and the correlation coefficient of the two samples from the same case was more than 0.99. When the modules were applied to 131 seized methamphetamine samples, four samples from two different cases were found to have the common origin and the chromatograms of the four samples were appeared visually identical

  20. Urine sample preparation in 96-well filter plates for quantitative clinical proteomics. (United States)

    Yu, Yanbao; Suh, Moo-Jin; Sikorski, Patricia; Kwon, Keehwan; Nelson, Karen E; Pieper, Rembert


    Urine is an important, noninvasively collected body fluid source for the diagnosis and prognosis of human diseases. Liquid chromatography mass spectrometry (LC-MS) based shotgun proteomics has evolved as a sensitive and informative technique to discover candidate disease biomarkers from urine specimens. Filter-aided sample preparation (FASP) generates peptide samples from protein mixtures of cell lysate or body fluid origin. Here, we describe a FASP method adapted to 96-well filter plates, named 96FASP. Soluble urine concentrates containing ~10 μg of total protein were processed by 96FASP and LC-MS resulting in 700-900 protein identifications at a 1% false discovery rate (FDR). The experimental repeatability, as assessed by label-free quantification and Pearson correlation analysis for shared proteins among replicates, was high (R ≥ 0.97). Application to urinary pellet lysates which is of particular interest in the context of urinary tract infection analysis was also demonstrated. On average, 1700 proteins (±398) were identified in five experiments. In a pilot study using 96FASP for analysis of eight soluble urine samples, we demonstrated that protein profiles of technical replicates invariably clustered; the protein profiles for distinct urine donors were very different from each other. Robust, highly parallel methods to generate peptide mixtures from urine and other body fluids are critical to increase cost-effectiveness in clinical proteomics projects. This 96FASP method has potential to become a gold standard for high-throughput quantitative clinical proteomics.

  1. Using Exclusion-Based Sample Preparation (ESP to Reduce Viral Load Assay Cost.

    Directory of Open Access Journals (Sweden)

    Scott M Berry

    Full Text Available Viral load (VL measurements are critical to the proper management of HIV in developing countries. However, access to VL assays is limited by the high cost and complexity of existing assays. While there is a need for low cost VL assays, performance must not be compromised. Thus, new assays must be validated on metrics of limit of detection (LOD, accuracy, and dynamic range. Patient plasma samples from the Joint Clinical Research Centre in Uganda were de-identified and measured using both an existing VL assay (Abbott RealTime HIV-1 and our assay, which combines low cost reagents with a simplified method of RNA isolation termed Exclusion-Based Sample Preparation (ESP.71 patient samples with VLs ranging from 3,000,000 copies/mL were used to compare the two methods. We demonstrated equivalent LOD (~50 copies/mL and high accuracy (average difference between methods of 0.08 log, R2 = 0.97. Using expenditures from this trial, we estimate that the cost of the reagents and consumables for this assay to be approximately $5 USD. As cost is a significant barrier to implementation of VL testing, we anticipate that our assay will enhance access to this critical monitoring test in developing countries.

  2. Using Exclusion-Based Sample Preparation (ESP) to Reduce Viral Load Assay Cost. (United States)

    Berry, Scott M; Pezzi, Hannah M; Williams, Eram D; Loeb, Jennifer M; Guckenberger, David J; Lavanway, Alex J; Puchalski, Alice A; Kityo, Cissy M; Mugyenyi, Peter N; Graziano, Franklin M; Beebe, David J


    Viral load (VL) measurements are critical to the proper management of HIV in developing countries. However, access to VL assays is limited by the high cost and complexity of existing assays. While there is a need for low cost VL assays, performance must not be compromised. Thus, new assays must be validated on metrics of limit of detection (LOD), accuracy, and dynamic range. Patient plasma samples from the Joint Clinical Research Centre in Uganda were de-identified and measured using both an existing VL assay (Abbott RealTime HIV-1) and our assay, which combines low cost reagents with a simplified method of RNA isolation termed Exclusion-Based Sample Preparation (ESP).71 patient samples with VLs ranging from 3,000,000 copies/mL were used to compare the two methods. We demonstrated equivalent LOD (~50 copies/mL) and high accuracy (average difference between methods of 0.08 log, R2 = 0.97). Using expenditures from this trial, we estimate that the cost of the reagents and consumables for this assay to be approximately $5 USD. As cost is a significant barrier to implementation of VL testing, we anticipate that our assay will enhance access to this critical monitoring test in developing countries.

  3. Magnetic hyperthermia heating of cobalt ferrite nanoparticles prepared by low temperature ferrous sulfate based method

    Directory of Open Access Journals (Sweden)

    Tejabhiram Yadavalli


    Full Text Available A facile low temperature co-precipitation method for the synthesis of crystalline cobalt ferrite nanostructures using ferrous sulfate salt as the precursor has been discussed. The prepared samples were compared with nanoparticles prepared by conventional co-precipitation and hydrothermal methods using ferric nitrate as the precursor. X-ray diffraction studies confirmed the formation of cubic spinel cobalt ferrites when dried at 110 °C as opposed to conventional methods which required higher temperatures/pressure for the formation of the same. Field emission scanning electron microscope studies of these powders revealed the formation of nearly spherical nanostructures in the size range of 20-30 nm which were comparable to those prepared by conventional methods. Magnetic measurements confirmed the ferromagnetic nature of the cobalt ferrites with low magnetic remanance. Further magnetic hyperthermia studies of nanostructures prepared by low temperature method showed a rise in temperature to 50 °C in 600 s.

  4. Sample processing and cDNA preparation for microbial metatranscriptomics in complex soil communities. (United States)

    Carvalhais, Lilia C; Schenk, Peer M


    Soil presents one of the most complex environments for microbial communities as it provides many microhabitats that allow coexistence of thousands of species with important ecosystem functions. These include biomass and nutrient cycling, mineralization, and detoxification. Culture-independent DNA-based methods, such as metagenomics, have revealed operational taxonomic units that suggest a high diversity of microbial species and associated functions in soil. An emerging but technically challenging area to profile the functions of microorganisms and their activities is mRNA-based metatranscriptomics. Here, we describe issues and important considerations of soil sample processing and cDNA preparation for metatranscriptomics from bacteria and archaea and provide a set of methods that can be used in the required experimental steps. © 2013 Elsevier Inc. All rights reserved.

  5. Summary Report for Evaluation of Compost Sample Drying Methods

    National Research Council Canada - National Science Library

    Frye, Russell


    .... Previous work in Support of these efforts developed a compost sample preparation scheme, consisting of air drying followed by milling, to reduce analytical variability in the heterogeneous compost matrix...

  6. Statistical sampling method, used in the audit

    Directory of Open Access Journals (Sweden)

    Gabriela-Felicia UNGUREANU


    Full Text Available The rapid increase in the size of U.S. companies from the early twentieth century created the need for audit procedures based on the selection of a part of the total population audited to obtain reliable audit evidence, to characterize the entire population consists of account balances or classes of transactions. Sampling is not used only in audit – is used in sampling surveys, market analysis and medical research in which someone wants to reach a conclusion about a large number of data by examining only a part of these data. The difference is the “population” from which the sample is selected, ie that set of data which is intended to draw a conclusion. Audit sampling applies only to certain types of audit procedures.

  7. [Preparation of clenbuterol monoclonal antibody with subtractive immunization method]. (United States)

    Li, Xiao-Li; Li, Xiao-Fang; Ning, Bao-An; Wu, Da-Cheng; Wang, Hong-Yong; Chen, Xiang; Ma, Xin-Hua; Ou, Guo-Rong; Gau, Zhi-Xian


    To obtain Clenbuterol monoclonal antibodies. Clenbuterol complete antigen was prepared with diazotization method. BALB/c mice was immunized with subtractive immunization, Clenbuterol monoclonal antibody was prepared with rule hybridoma technique. The mice obtained tolerance to BSA by subtractive immunization. The rate of the hybridoma cell with positive reaction which had obtained was 8.2%, and the specific clenbuterol monoclonal antibody was obtained at last. Monoclonal antibodies to micromolecule contaminant be prepared by subtractive immunization, could decrease the workload in the bolting of monoclonal antibodies, and increase the chance to obtain the antibody of expected.

  8. An improved method of preparation of nanoparticular metal oxide catalysts

    DEFF Research Database (Denmark)


    The present invention concerns an improved method of preparation of nanoparticular vanadium oxide/anatase titania catalysts having a narrow particle size distribution. In particular, the invention concerns preparation of nanoparticular vanadium oxide/anatase titania catalyst precursors comprising...... combustible crystallization seeds upon which the catalyst metal oxide is coprecipitated with the carrier metal oxide, which crystallization seeds are removed by combustion in a final calcining step....

  9. pH adjustment of human blood plasma prior to bioanalytical sample preparation

    NARCIS (Netherlands)

    Hendriks, G.; Uges, D. R. A.; Franke, J. P.


    pH adjustment in bioanalytical sample preparation concerning ionisable compounds is one of the most common sample treatments. This is often done by mixing an aliquot of the sample with a proper buffer adjusted to the proposed pH. The pH of the resulting mixture however, does not necessarily have to

  10. Implementation of an office-based semen preparation method (SEP ...

    African Journals Online (AJOL)

    Implementation of an office-based semen preparation method (SEP-D Kit) for intra-uterine insemination (IUI): A controlled randomised study to compare the IUI pregnancy outcome between a routine (swim-up) and the SEP-D Kit method.

  11. Squalane and isosqualane compositions and methods for preparing the same (United States)

    Fisher, Karl; Schofer, Susan Jessica; Kanne, David B


    Provided herein are methods comprising catalytic dimerization of .beta.-farnesene to obtain squalane and/or isosqualane. Compositions comprising squalane and/or isosqualane are provided. In certain embodiments, squalane and isosqualane prepared by the methods provided herein can be useful for applications in cosmetic industry and/or in the lubricants industry.

  12. Sample preparation and UHPLC-FD analysis of pteridines in human urine. (United States)

    Tomšíková, H; Solich, P; Nováková, L


    Elevated levels of pteridines can indicate the activation of cellular immune system by certain diseases. No work dealing with the simultaneous determination of urinary neopterin, biopterin and their reduced forms has been published. Therefore, a new SPE-UHPLC-FD method for the analysis of these compounds has been developed. The main emphasis was put on the stability of dihydroforms during the sample processing and storage. As a stabilizing agent, dithiothreitol, at various concentrations, and various pH values (3.8-9.8) of working solutions were tested. Chromatographic separation was performed under HILIC isocratic conditions on BEH Amide column. The method was linear for the calibration standard solutions in the range of 10-10,000 ng/ml (dihydroforms) and 0.5-1000 ng/ml (oxidized forms), and for real samples in the range of 25-1000 ng/ml (dihydroforms) and 1-100 ng/ml (oxidized forms). The development of a new SPE sample preparation method was carried out on different types of sorbents (based on a mixed-mode cation exchange, porous graphitic carbon and a polymer comprising hydrophilic and hydrophobic components). Final validation was performed on a MCAX SPE column. Method accuracy ranged from 76.9 to 121.9%. The intra- and inter-day precision did not exceed 10.7%. The method provided high sensitivity for the use in routine clinical measurements of urine (LLOQ 1 ng/ml for oxidized forms and 25 ng/ml for dihydroforms). Average concentrations of biopterin, neopterin, and dihydrobiopterin found in urine of healthy persons were related to the mol of creatinine (66.8, 142.3, and 257.3 μmol/mol of creatinine, respectively) which corresponded to the literature data. The concentration of dihydroneopterin obtained using our method was 98.8 μmol/mol of creatinine. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Protocol: MYTHBUSTERS: a universal procedure for sample preparation for mass spectrometry. (United States)

    Drabik, Anna; Ner-Kluza, Joanna; Bodzon-Kulakowska, Anna; Suder, Piotr

    Improvements in proteomic strategies from the development of new and robust separation and identification techniques have led to broad applications of proteomics to solve numerous biological questions. For all analyses, sample quality is unquestionably a critical factor; therefore protein extraction is of outmost importance. The ideal extraction method should provide reproducible spectra of the most comprehensive repertoire of proteins, while minimizing sample loss and degradation. It is already known that to capture the whole proteome is an unenforceable task. Many protein extraction protocols have been described, yet there is no "one perfect procedure" taking into account the vast diversity of biological and physical properties of proteins, including their charge, size, hydrophobicity, interactions and sub-cellular localization. The research presented here reflects the main obstacle occurring in proteomic experimental design; i.e. the lack of reproducibility as a result of alterations in protein extraction methods. We have performed a series of experiments, aimed towards identification of the aptamer-binding partners in cancerous cells. Aptamers are chemically synthesized, short, single-stranded nucleic acids with a strictly defined three-dimensional structure, which allows them to interact with a target molecule with high affinity. The low immunogenicity and cellular- targeting properties of aptamers might facilitate design of suitable drugs with low side-effects. Aptamers can be used for identification of molecules associated with a pathogenic state of a cell. Aptamers can be considered as a powerful tool, since they possess unique properties to benefit cancer diagnosis, prevention and treatment. We have used different types of protein extraction methods prior to analyses of complex biological samples by mass spectrometry, based on slight changes of homogenization buffers, and have observed the changes in the identified compounds. These results should prove to

  14. Matrix compatible solid phase microextraction coating, a greener approach to sample preparation in vegetable matrices. (United States)

    Naccarato, Attilio; Pawliszyn, Janusz


    This work proposes the novel PDMS/DVB/PDMS fiber as a greener strategy for analysis by direct immersion solid phase microextraction (SPME) in vegetables. SPME is an established sample preparation approach that has not yet been adequately explored for food analysis in direct immersion mode due to the limitations of the available commercial coatings. The robustness and endurance of this new coating were investigated by direct immersion extractions in raw blended vegetables without any further sample preparation steps. The PDMS/DVB/PDMS coating exhibited superior features related to the capability of the external PDMS layer to protect the commercial coating, and showed improvements in terms of extraction capability and in the cleanability of the coating surface. In addition to having contributed to the recognition of the superior features of this new fiber concept before commercialization, the outcomes of this work serve to confirm advancements in the matrix compatibility of the PDMS-modified fiber, and open new prospects for the development of greener high-throughput analytical methods in food analysis using solid phase microextraction in the near future. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. A New Method for Preparation of Metal Matrix Nanocomposites (United States)

    Padhi, Payodhar; Panigrahi, S. C.; Ghosh, Sudipto


    Particulate metal matrix composites (MMCs) can involve ceramic particulates ranging in size from few nanometers to 500 μm. Particulates are added to the metal matrix for strengthening. In particular, addition of nanoparticles, even in quantities as small as 2 weight percent can enhance the hardness or yield strength by a factor as high as 2. There are several methods for the production of metal matrix nanocomposites including mechanical alloying , vertex process and spray deposition. However, the above processes are expensive. Solidification processing is a relatively cheaper route. However, during solidification processing nanoparticulates tend to agglomerate as a result of van der Waals forces and thus proper dispersion of the nano-particulate in metal matrix is a challenge. Yang et al dispersed nanoparticles in metal matrix by ultrasonic casting. However their technique has several drawbacks such as the oscillating probe, which is in direct contact with liquid metal, may dissolve in the liquid metal and contaminate it. Moreover, the extent of dispersion is not uniform. It is maximum near the probe and gradually decreases as one move away from the probe. Lastly in the method developed by Yang et al, the oscillating probe is removed from the liquid metal before cooling and solidification begin. This may lead to partial reagglomeration of nanoparticles. To overcome these difficulties a non-contact method, where the ultrasonic probe is not in direct contact with the liquid metal, was attempted to disperse nano-sized Al2O3 particulates in aluminum matrix. In this method the mold was subjected to ultrasonic vibration. Hardness measurements and microstructural studies using HRTEM were carried out on samples taken from different locations of the nanocomposite ingot cast by the non-contact method. Commercially pure liquid aluminum was used as matrix of the composite. The Al2O3 nano-powder was prepared by ball milling for 22 hr. The nanopowders were characterized using

  16. Study of the first paramagnetic to ferromagnetic transition in as prepared samples of Mn–Fe–P–Si magnetocaloric compounds prepared by different synthesis routes

    Energy Technology Data Exchange (ETDEWEB)

    Bartok, A., E-mail: [SATIE, ENS Cachan, CNRS, Universite Paris-Saclay, 61 av President Wilson, F-94230 Cachan (France); Kustov, M.; Cohen, L.F. [Imperial College, London SW7 2AZ (United Kingdom); Pasko, A. [SATIE, ENS Cachan, CNRS, Universite Paris-Saclay, 61 av President Wilson, F-94230 Cachan (France); Zehani, K.; Bessais, L. [CMTR, ICMPE, CNRS-UPEC, 2-8 rue Henri Dunant, F-94320 Thiais (France); Mazaleyrat, F.; LoBue, M. [SATIE, ENS Cachan, CNRS, Universite Paris-Saclay, 61 av President Wilson, F-94230 Cachan (France)


    Magnetocaloric materials with composition of Mn{sub 1.3}Fe{sub 0.65}P{sub 0.5} Si{sub 0.5} have been prepared by ball milling and solid-state reaction methods and consolidated using powder annealing, and conventional and spark plasma sintering. Magnetic and calorimetric measurements show remarkable differences upon first cooling, and slight differences on second and further coolings between the samples prepared by different synthesis routes. Further measurements using Hall probe imaging in high magnetic field have been also carried out. As-prepared samples have been cooled down just above the critical temperature, and the first phase transition has been induced by application of a magnetic field. Bulk samples show staircase isothermal magnetization curves whereas powders show smoother transition curves. - Highlights: • Mn–Fe–P–Si magnetocaloric materials have been prepared by different synthesis routes. • Magnetic measurements show remarkable differences upon first cooling. • First phase transition has been induced by application of a magnetic field. • Hall probe imaging in high magnetic field has also been carried out. • Bulk samples crack during the first PM–FM transition.

  17. Simple and Reproducible Sample Preparation for Single-Shot Phosphoproteomics with High Sensitivity

    DEFF Research Database (Denmark)

    Jersie-Christensen, Rosa R.; Sultan, Abida; Olsen, Jesper V


    The traditional sample preparation workflow for mass spectrometry (MS)-based phosphoproteomics is time consuming and usually requires multiple steps, e.g., lysis, protein precipitation, reduction, alkylation, digestion, fractionation, and phosphopeptide enrichment. Each step can introduce chemica...

  18. Robotic, MEMS-based Multi Utility Sample Preparation Instrument for ISS Biological Workstation Project (United States)

    National Aeronautics and Space Administration — This project will develop a multi-functional, automated sample preparation instrument for biological wet-lab workstations on the ISS. The instrument is based on a...

  19. In-Situ Sample Preparation Development for Extraterrestrial Life Detection and Characterization (United States)

    Craft, K. L.; Bradburne, C.; Tiffany, J.; Hagedon, M.; Hibbitts, C.; Vandegriff, J.; Horst, S.


    In-situ life detection instrumentation require robust sample preparation techniques that need further development in the coming years to enable the exciting life discoveries we seek in both familiar and unfamiliar planetary environments.

  20. Development of automated preparation system for isotopocule analysis of N2O in various air samples (United States)

    Toyoda, Sakae; Yoshida, Naohiro


    Nitrous oxide (N2O), an increasingly abundant greenhouse gas in the atmosphere, is the most important stratospheric ozone-depleting gas of this century. Natural abundance ratios of isotopocules of N2O, NNO molecules substituted with stable isotopes of nitrogen and oxygen, are a promising index of various sources or production pathways of N2O and of its sink or decomposition pathways. Several automated methods have been reported to improve the analytical precision for the isotopocule ratio of atmospheric N2O and to reduce the labor necessary for complicated sample preparation procedures related to mass spectrometric analysis. However, no method accommodates flask samples with limited volume or pressure. Here we present an automated preconcentration system which offers flexibility with respect to the available gas volume, pressure, and N2O concentration. The shortest processing time for a single analysis of typical atmospheric sample is 40 min. Precision values of isotopocule ratio analysis are automated systems, but better than that of our previously reported manual measurement system.

  1. Preparation of well-defined samples of AlPdMn quasicrystals for surface studies (United States)

    Jenks, C. J.; Delaney, D. W.; Bloomer, T. E.; Chang, S.-L.; Lograsso, T. A.; Shen, Z.; Zhang, C.-M.; Thiel, P. A.


    We have developed a method for preparing single-grain, quasicrystalline AlPdMn samples for surface studies in ultrahigh vacuum. The main issues of concern are phase purity, the quality of the surface structure, and the surface, and the surface composition. Phase purity is enhanced by annealing the sample in ultra-pure Ar in a sealed quartz ampoule for several days before polishing. Polishing with colloidal silica allows secondary phases to be detected readily with an optical microscope. As a final precaution, phase purity can be checked sensitively with scanning Auger microscopy. After this stage, the sample can be cleaned in ultrahigh vacuum with ion bombardment. Annealing is required after bombardment to restore surface structure and to obtain a low-energy electron diffraction (LEED) pattern of an oriented sample. However, both ion bombardment and heating to temperatures above 870 K in vacuum, produce Pd-rich surfaces. As a final step, for the five-fold surface, we recommend heating briefly to 1050-1100 K and then annealing at 870 K for several hours. This produces both an excellent LEED pattern, and a surface composition close to that of the bulk.

  2. Determination of bromine, fluorine and iodine in mineral supplements using pyrohydrolysis for sample preparation

    Energy Technology Data Exchange (ETDEWEB)

    Taflik, Ticiane; Antes, Fabiane G.; Paniz, Jose N.G.; Flores, Erico M.M.; Dressler, Valderi L., E-mail: [Departamento de Quimica, Universidade Federal de Santa Maria, RS (Brazil); Duarte, Fabio A. [Escola de Quimica e Alimentos, Universidade Federal do Rio Grande, Rio Grande, RS (Brazil); Flores, Eder L.M. [Coordenacao de Engenharia de Alimentos, Universidade Tecnologica Federal do Parana, Medianeira, PR (Brazil)


    Pyrohydrolysis was employed for mineral supplements decomposition prior to F, Br and I determination. Fluoride determination was carried out by potentiometry using a fluoride-ion selective electrode, whereas Br and I were determined by inductively coupled plasma mass spectrometry. The main parameters that influence on pyrohydrolysis were investigated. After evaluation, the following conditions were established: reactor temperature of 1000 deg C during 10 min; sample plus accelerator mass ratio of 1 + 5 and carrier gas (air) flow rate of 200 mL min{sup -1} . The accuracy of the proposed method was evaluated by analyte recovery tests and analysis of certified reference materials of phosphate rock and soil. Commercial mineral supplement samples were analyzed. The limits of quantification were 16, 0.3 and 0.07 {mu}g g{sup -1} for F, Br and I, respectively. By using a relatively simple and low cost pyrohydrolysis system up to 5 samples can be processed per hour. The developed sample preparation procedure can be routinely employed for F, Br and I determination in mineral supplements. (author)

  3. Preparation

    Directory of Open Access Journals (Sweden)

    M.M. Dardir


    Full Text Available Some hexanamide-mono and di-linoleniate esters were prepared by the reaction of linolenic acid and hexanamide (derived from the reaction of hexanoic acid and diethanolamine. The chemical structure for the newly prepared hexanamide-mono and di-linoleniate esters were elucidated using elemental analysis, (FTIR, H 1NMR and chemical ionization mass spectra (CI/Ms spectroscopic techniques. The results of the spectroscopic analysis indicated that they were prepared through the right method and they have high purity. The new prepared esters have high biodegradability and lower toxicity (environmentally friendly so they were evaluated as a synthetic-based mud (ester-based mud for oil-well drilling fluids. The evaluation included study of the rheological properties, filtration and thermal properties of the ester based-muds formulated with the newly prepared esters compared to the reference commercial synthetic-based mud.

  4. Optimized sample preparation for MALDI mass spectrometry analysis of protected synthetic peptides. (United States)

    Schaiberger, Audrey M; Moss, Jason A


    The recent development and commercialization of Fuzeon (enfuvirtide) demonstrated that a convergent strategy comprised of both solid- and solution-phase synthetic methodologies presents a viable route for peptide manufacturing on a multi-ton scale. In this strategy, the target sequence is prepared by stepwise solid-phase synthesis of protected peptide fragments, which are then coupled together in the solution-phase to give the full-length sequence. These synthetic methodologies pose a unique challenge for mass spectrometry (MS), as protected peptide intermediates are often marked by poor solubility, structural lability, and low ionization potential. Matrix-assisted laser desorption/ionization (MALDI) MS is uniquely suited to such analytes; however, generalized protocols for MALDI analysis of protected peptides have yet to be demonstrated. Herein, we report an operationally simple sample preparation method for MALDI analysis of protected peptides, which greatly facilitates the collection and interpretation of MS data. In this method, the difficulty in MS analysis of protected peptides has been greatly diminished by use of dithranol as a matrix and CsCl as an additive, giving rise to intentionally-formed Cs(+) adducts. With greatly reduced fragmentation, better crystalline morphology, and easier data interpretation, we anticipate that these findings will find utility in peptide process development and manufacturing settings for reaction monitoring, troubleshooting, and quality control.

  5. Sample Preparation (SS) - Metabolonote | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data List Contact us Metabol...ed to by S (Sample information). Data file File name: File URL: File size: 13 KB Simple search URL Download License Update History of This Database Site Policy | Contact Us Sample Preparation (SS) - Metabolonote | LSDB Archive ...

  6. The structural properties of barium cobalt hexaferrite powder prepared by a simple heat treatment method

    Energy Technology Data Exchange (ETDEWEB)

    Chauhan, Chetna, E-mail: [Electrical Engineering Department, Institute of Technology, Nirma University, Ahmedabad-382 481. Gujarat. India (India); Jotania, Rajshree, E-mail: [Department of Physics, University School of Sciences, Gujarat University, Ahmeabad – 380009. Gujarat. India (India)


    The W-type barium hexaferrite was prepared using a simple heat treatment method. The precursor was calcinated at 650°C for 3 hours and then slowly cooled to room temperature in order to obtain barium cobalt hexaferrite powder. The prepared powder was characterised by different experimental techniques like XRD, FTIR and SEM. The X-ray diffractogram of the sample shows W-and M phases. The particle size calculated by Debye Scherrer formula. The FTIR spectra of the sample was taken at room temperature by using KBr pallet method which confirms the formation of hexaferrite phase. The morphological study on the hexaferrite powder was carried out by SEM analysis.

  7. The structural properties of barium cobalt hexaferrite powder prepared by a simple heat treatment method (United States)

    Chauhan, Chetna; Jotania, Rajshree


    The W-type barium hexaferrite was prepared using a simple heat treatment method. The precursor was calcinated at 650°C for 3 hours and then slowly cooled to room temperature in order to obtain barium cobalt hexaferrite powder. The prepared powder was characterised by different experimental techniques like XRD, FTIR and SEM. The X-ray diffractogram of the sample shows W-and M phases. The particle size calculated by Debye Scherrer formula. The FTIR spectra of the sample was taken at room temperature by using KBr pallet method which confirms the formation of hexaferrite phase. The morphological study on the hexaferrite powder was carried out by SEM analysis.

  8. Comparison between powder and slices diffraction methods in teeth samples

    Energy Technology Data Exchange (ETDEWEB)

    Colaco, Marcos V.; Barroso, Regina C. [Universidade do Estado do Rio de Janeiro (IF/UERJ), RJ (Brazil). Inst. de Fisica. Dept. de Fisica Aplicada; Porto, Isabel M. [Universidade Estadual de Campinas (FOP/UNICAMP), Piracicaba, SP (Brazil). Fac. de Odontologia. Dept. de Morfologia; Gerlach, Raquel F. [Universidade de Sao Paulo (FORP/USP), Rieirao Preto, SP (Brazil). Fac. de Odontologia. Dept. de Morfologia, Estomatologia e Fisiologia; Costa, Fanny N. [Coordenacao dos Programas de Pos-Graduacao de Engenharia (LIN/COPPE/UFRJ), RJ (Brazil). Lab. de Instrumentacao Nuclear


    Propose different methods to obtain crystallographic information about biological materials are important since powder method is a nondestructive method. Slices are an approximation of what would be an in vivo analysis. Effects of samples preparation cause differences in scattering profiles compared with powder method. The main inorganic component of bones and teeth is a calcium phosphate mineral whose structure closely resembles hydroxyapatite (HAp). The hexagonal symmetry, however, seems to work well with the powder diffraction data, and the crystal structure of HAp is usually described in space group P63/m. Were analyzed ten third molar teeth. Five teeth were separated in enamel, detin and circumpulpal detin powder and five in slices. All the scattering profile measurements were carried out at the X-ray diffraction beamline (XRD1) at the National Synchrotron Light Laboratory - LNLS, Campinas, Brazil. The LNLS synchrotron light source is composed of a 1.37 GeV electron storage ring, delivering approximately 4x10{sup -1}0 photons/s at 8 keV. A double-crystal Si(111) pre-monochromator, upstream of the beamline, was used to select a small energy bandwidth at 11 keV . Scattering signatures were obtained at intervals of 0.04 deg for angles from 24 deg to 52 deg. The human enamel experimental crystallite size obtained in this work were 30(3)nm (112 reflection) and 30(3)nm (300 reflection). These values were obtained from measurements of powdered enamel. When comparing the slice obtained 58(8)nm (112 reflection) and 37(7)nm (300 reflection) enamel diffraction patterns with those generated by the powder specimens, a few differences emerge. This work shows differences between powder and slices methods, separating characteristics of sample of the method's influence. (author)

  9. Closer to the native state. Critical evaluation of cryo-techniques for Transmission Electron Microscopy: preparation of biological samples. (United States)

    Mielanczyk, Lukasz; Matysiak, Natalia; Michalski, Marek; Buldak, Rafal; Wojnicz, Romuald


    Over the years Transmission Electron Microscopy (TEM) has evolved into a powerful technique for the structural analysis of cells and tissues at various levels of resolution. However, optimal sample preservation is required to achieve results consistent with reality. During the last few decades, conventional preparation methods have provided most of the knowledge about the ultrastructure of organelles, cells and tissues. Nevertheless, some artefacts can be introduced at all stagesofstandard electron microscopy preparation technique. Instead, rapid freezing techniques preserve biological specimens as close as possible to the native state. Our review focuses on different cryo-preparation approaches, starting from vitrification methods dependent on sample size. Afterwards, we discuss Cryo-Electron Microscopy Of VItreous Sections (CEMOVIS) and the main difficulties associated with this technique. Cryo-Focused Ion Beam (cryo-FIB) is described as a potential alternative for CEMOVIS. Another post-processing route for vitrified samples is freeze substitution and embedding in resin for structural analysis or immunolocalization analysis. Cryo-sectioning according to Tokuyasu is a technique dedicated to high efficiency immunogold labelling. Finally, we introduce hybrid techniques, which combine advantages of primary techniques originally dedicated to different approaches. Hybrid approaches permit to perform the study of difficult-to-fix samples and antigens or help optimize the sample preparation protocol for the integrated Laser and Electron Microscopy (iLEM) technique.

  10. Evaluation of Six Sample Preparation Methods for Determination of ...

    African Journals Online (AJOL)


    A MARS 5 microwave digestion system (CEM Corporation,. USA) was employed for lubricating oil digestion. It was equipped with ESP-1500 plus pressure and RTP-300 plus temperature control systems. Teflon EasyPreps vessels, allowing a maximum decomposition pressure of 800 psi at 240 °C were used to digest.

  11. Comparison of sample preparation methods in toxicological analysis


    Dulius, Aurimas


    Šiame darbe nagrinėjami kietazės ekstrakcijos ir skysčių – skysčių ekstrakcijos metodai, bei lyginami jų efektyvumai ekstrahuojant kodeiną iš kraujo plazmos ir šlapimo. Ekstrakcijų efektyvumai buvo nustatinėjami UV spektrometrijos būdu. Darbo tikslas – nustatyti optimalias mėginio ruošimo toksikologinėje analizėje metodikas, tinkamas kodeino išskyrimui iš biologinių terpių – kraujo plazmos, šlapimo, bei palyginti šių metodikų efektyvumą. įvertinus gautus duomenis, galima teigti, kad ekstrahuo...

  12. Development of a Modified Smart System for Robust Transcriptome Library Preparation from Limited Quantities of Compromised Samples (United States)

    Chang, C.; Farmer, A.; Bostick, Magnolia


    Next Generation Sequencing has revolutionized biomedical research by providing sequence data on millions of short DNA fragments, in parallel. In particular, NGS has enabled RNA expression analysis over the entire transcriptome with high sensitivity and dynamic range. Currently, the field is seeking methods to utilize challenging samples that are either compromised or are only available in limited amounts. Overcoming these constraints will demand highly sensitive and robust sample preparation methods. One powerful method for cDNA preparation is SMART™ technology (Switching Mechanism At the 5′ end of the RNA Template), which utilizes the template switching activity of reverse transcriptase to enable the direct addition of a PCR adaptor to the 3′ end of the first-strand cDNA, thus avoiding inefficient ligation steps. One drawback of SMART technology is its current inability to work with compromised samples, owing to its dependence on an oligo dT primer for first strand synthesis. A modified SMART system has been developed including the use of random primers to work with samples containing compromised or degraded RNA. Data (including gene body coverage, reproducibility, and mappability metrics) will be presented for both chemically degraded RNA samples and FFPE RNA prepared using the modified SMART system. This modified SMART protocol will be especially useful for small samples of degraded RNA. It is capable of generating cDNA libraries for transcriptome profiling from as little as 1 ng of total RNA.

  13. M3FT-17OR0301070211 - Preparation of Hot Isostatically Pressed AgZ Waste Form Samples

    Energy Technology Data Exchange (ETDEWEB)

    Jubin, Robert Thomas [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Bruffey, Stephanie H. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Jordan, Jacob A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)


    The production of radioactive iodine-bearing waste forms that exhibit long-term stability and are suitable for permanent geologic disposal has been the subject of substantial research interest. One potential method of iodine waste form production is hot isostatic pressing (HIP). Recent studies at Oak Ridge National Laboratory (ORNL) have investigated the conversion of iodine-loaded silver mordenite (I-AgZ) directly to a waste form by HIP. ORNL has performed HIP with a variety of sample compositions and pressing conditions. The base mineral has varied among AgZ (in pure and engineered forms), silver-exchanged faujasite, and silverexchanged zeolite A. Two iodine loading methods, occlusion and chemisorption, have been explored. Additionally, the effects of variations in temperature and pressure of the process have been examined, with temperature ranges of 525°C–1,100°C and pressure ranges of 100–300 MPa. All of these samples remain available to collaborators upon request. The sample preparation detailed in this document is an extension of that work. In addition to previously prepared samples, this report documents the preparation of additional samples to support stability testing. These samples include chemisorbed I-AgZ and pure AgI. Following sample preparation, each sample was processed by HIP by American Isostatic Presses Inc. and returned to ORNL for storage. ORNL will store the samples until they are requested by collaborators for durability testing. The sample set reported here will support waste form durability testing across the national laboratories and will provide insight into the effects of varied iodine content on iodine retention by the produced waste form and on potential improvements in waste form durability provided by the zeolite matrix.

  14. System and method for extracting a sample from a surface (United States)

    Van Berkel, Gary; Covey, Thomas


    A system and method is disclosed for extracting a sample from a sample surface. A sample is provided and a sample surface receives the sample which is deposited on the sample surface. A hydrophobic material is applied to the sample surface, and one or more devices are configured to dispense a liquid on the sample, the liquid dissolving the sample to form a dissolved sample material, and the one or more devices are configured to extract the dissolved sample material from the sample surface.

  15. Perpendicular distance sampling: an alternative method for sampling downed coarse woody debris (United States)

    Michael S. Williams; Jeffrey H. Gove


    Coarse woody debris (CWD) plays an important role in many forest ecosystem processes. In recent years, a number of new methods have been proposed to sample CWD. These methods select individual logs into the sample using some form of unequal probability sampling. One concern with most of these methods is the difficulty in estimating the volume of each log. A new method...

  16. Magnetic separation techniques in sample preparation for biological analysis: a review. (United States)

    He, Jincan; Huang, Meiying; Wang, Dongmei; Zhang, Zhuomin; Li, Gongke


    Sample preparation is a fundamental and essential step in almost all the analytical procedures, especially for the analysis of complex samples like biological and environmental samples. In past decades, with advantages of superparamagnetic property, good biocompatibility and high binding capacity, functionalized magnetic materials have been widely applied in various processes of sample preparation for biological analysis. In this paper, the recent advancements of magnetic separation techniques based on magnetic materials in the field of sample preparation for biological analysis were reviewed. The strategy of magnetic separation techniques was summarized. The synthesis, stabilization and bio-functionalization of magnetic nanoparticles were reviewed in detail. Characterization of magnetic materials was also summarized. Moreover, the applications of magnetic separation techniques for the enrichment of protein, nucleic acid, cell, bioactive compound and immobilization of enzyme were described. Finally, the existed problems and possible trends of magnetic separation techniques for biological analysis in the future were proposed. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Isotachophoretic phenomena in electric field gradient focusing: perspectives for sample preparation and bioassays. (United States)

    Quist, Jos; Vulto, Paul; Hankemeier, Thomas


    Isotachophoresis (ITP) and electric field gradient focusing (EFGF) are two powerful approaches for simultaneous focusing and separation of charged compounds. Remarkably, in many EFGF methods, isotachophoretic hallmarks have been found, including observations of plateau concentrations and contiguous analyte bands. We discuss the similarities between ITP and EFGF and describe promising possibilities to transfer the functionality and applications developed on one platform to other platforms. Of particular importance is the observation that single-electrolyte isotachophoretic separations with tunable ionic mobility window can be performed, as is illustrated with the example of depletion zone isotachophoresis (dzITP). By exploiting the rapid developments in micro- and nanofluidics, many interesting combinations of ITP and EFGF features can be achieved, yielding powerful analytical platforms for sample preparation, biomarker discovery, molecular interaction assays, drug screening, and clinical diagnostics.

  18. NGSI FY15 Final Report. Innovative Sample Preparation for in-Field Uranium Isotopic Determinations

    Energy Technology Data Exchange (ETDEWEB)

    Yoshida, Thomas M. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Meyers, Lisa [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)


    Our FY14 Final Report included an introduction to the project, background, literature search of uranium dissolution methods, assessment of commercial off the shelf (COTS) automated sample preparation systems, as well as data and results for dissolution of bulk quantities of uranium oxides, and dissolution of uranium oxides from swipe filter materials using ammonium bifluoride (ABF). Also, discussed were reaction studies of solid ABF with uranium oxide that provided a basis for determining the ABF/uranium oxide dissolution mechanism. This report details the final experiments for optimizing dissolution of U3O8 and UO2 using ABF and steps leading to development of a Standard Operating Procedure (SOP) for dissolution of uranium oxides on swipe filters.

  19. Evaluation of preparation methods for MS-based analysis of intestinal epithelial cell proteomes

    DEFF Research Database (Denmark)

    Hesselager, Marianne Overgaard; Codrea, Marius Cosmin; Bendixen, Emøke


    The gut epithelium formed between an organism and the environment plays an essential role in host–microbe interactions, yet remains one of the least characterized mammalian tissues. Especially the membrane proteins, which are critical to bacterial adhesion, are understudied, because these proteins...... are low in abundance, and large amounts of sample is needed for their preparation and for undertaking MS-based analysis. The aim of this study was to evaluate three different methods for isolation and preparation of pig intestinal epithelial cells for MS-based analysis of the proteome. Samples were...... of ease and speed of sample preparation, as well as protein recovery. In comparison, more gentle methods where intestinal epithelial cells are harvested by shaking are more time consuming, result in lower protein yield, and are prone to increased technical variation due to multiple steps involved....


    Directory of Open Access Journals (Sweden)

    M. I. Egorova


    Full Text Available Summary. Molasses is characterized as sugar production by-product from primary or secondary sacchariferous raw materials. The features of the appearance, the chemical composition, molasses and exit directions of its use, depending on the type of production, in which it is formed. The value of molasses is demonstrated according to its total composition as well as its use directions. Statistics on beet molasses amounts in Russia is presented. Described consumer market molasses in Russia and abroad with its exports. Shown regulations contain requirements for the quality and safety of molasses, including sulfur dioxide. The data on sulfur allergenic properties are presented. Showing source of the sulfur dioxide in the residual molasses number of processing aids and the impact of its level in the value of raw molasses for use in biotechnological processes and fodder production. The necessity to develop methodology for determining the sulfur dioxide content in the molasses to control its security. The iodometric method, which is used in practice for determination of sulphur dioxide in foods are characterized. Differences molasses and sugar as objects of iodometric determination of sulfur dioxide, which leads to the inability to ascertain the equivalence point. The variants eliminate interfering background of dark-colored foods common in analytical chemistry. Advantages and disadvantages of the background masking and stripping the determination of sulfur dioxide in the darkcolored products. It was characterized by clarifying sugar solutions in optical control methods. The hypothesis about preferability of its use in sample molasses preparation for equivalence point fixation in iodometric titration is suggested. The tasks of experimental research for the development of sample preparation algorithm molasses in determining the content of sulphurous acid.

  1. Influence of preparation method on structural and magnetic ...

    Indian Academy of Sciences (India)

    Nickel ferrite nanoparticles of very small size were prepared by sol–gel combustion and co-precipitation techniques. At the same annealing temperature sol–gel derived particles had bigger crystallite size. In both methods, crystallite size of the particles increased with annealing temperature. Sol–gel derived nickel ferrite ...

  2. Methods of preparation and the energy, protein and mineral values ...

    African Journals Online (AJOL)

    This study analyses the different methods of preparation and the nutritional value of three Cameroonian dishes. These are: "Corn chaff", "Nnam Owondo/Ebobolo" and "Nnam Ngon/Ebobolo". "Corn chaff" is a mixture of corn (Zea mays L.), beans (Phaseolus vulgaris) and palm oil (Elaeis guinensis). "Nnam Owondo/Ebobolo" ...

  3. The influence of preparation method on children's liking for vegetables

    NARCIS (Netherlands)

    Zeinstra, G.G.; Koelen, M.; Kok, F.J.; Graaf, de C.


    This study aimed to investigate how different preparation methods influence children’s liking for vegetables. Participants were children from three age groups (4–6 years N = 46; 7–8 years N = 25; 11–12 years N = 23) and young adults (18–25 years N = 22). The participants tasted and ranked six

  4. Method of preparing cross-linked enzyme particles

    NARCIS (Netherlands)

    Mateo, C.; Van Langen, L.M.; Van Rantwijk, F.


    The invention relates to a method of preparing cross-linked enzyme particles using a cross-linking agent. According to the invention, the enzyme particles are formed and subsequently cross-linked using a cross-linking agent having at least n reactive groups where N>=3 and a molecular weight of

  5. Modular approach to customise sample preparation procedures for viral metagenomics: a reproducible protocol for virome analysis. (United States)

    Conceição-Neto, Nádia; Zeller, Mark; Lefrère, Hanne; De Bruyn, Pieter; Beller, Leen; Deboutte, Ward; Yinda, Claude Kwe; Lavigne, Rob; Maes, Piet; Van Ranst, Marc; Heylen, Elisabeth; Matthijnssens, Jelle


    A major limitation for better understanding the role of the human gut virome in health and disease is the lack of validated methods that allow high throughput virome analysis. To overcome this, we evaluated the quantitative effect of homogenisation, centrifugation, filtration, chloroform treatment and random amplification on a mock-virome (containing nine highly diverse viruses) and a bacterial mock-community (containing four faecal bacterial species) using quantitative PCR and next-generation sequencing. This resulted in an optimised protocol that was able to recover all viruses present in the mock-virome and strongly alters the ratio of viral versus bacterial and 16S rRNA genetic material in favour of viruses (from 43.2% to 96.7% viral reads and from 47.6% to 0.19% bacterial reads). Furthermore, our study indicated that most of the currently used virome protocols, using small filter pores and/or stringent centrifugation conditions may have largely overlooked large viruses present in viromes. We propose NetoVIR (Novel enrichment technique of VIRomes), which allows for a fast, reproducible and high throughput sample preparation for viral metagenomics studies, introducing minimal bias. This procedure is optimised mainly for faecal samples, but with appropriate concentration steps can also be used for other sample types with lower initial viral loads.

  6. Fecal specimens preparation methods for PCR diagnosis of human taeniosis

    Directory of Open Access Journals (Sweden)

    Nunes Cáris Maroni


    Full Text Available Sample preparation and DNA extraction protocols for DNA amplification by PCR, which can be applied in human fecal samples for taeniasis diagnosis, are described. DNA extracted from fecal specimens with phenol/chloroform/isoamilic alcohol and DNAzol® reagent had to be first purified to generate fragments of 170 pb and 600 pb by HDP2-PCR. This purification step was not necessary with the use of QIAmp DNA stool mini kit®. Best DNA extraction results were achieved after eggs disruption with glass beads, either with phenol/chloroform/isoamilic alcohol, DNAzol® reagent or QIAmp DNA stool mini kit®.

  7. Sample preparation vs quality of X-ray phase analysis results

    Directory of Open Access Journals (Sweden)

    R. Chylińska


    Full Text Available In this study, taking as an example the creep-resistant austenitic cast steel, the results of the investigations were presented whose aim was to show what effect the specimen surface condition, discussed in terms of its roughness obtained by grinding, polishing with diamond paste, electrolytic polishing and etching, may have on the quality of results obtained by X-ray phase analysis. The preset goal has been achieved comparing the quantity and intensity of reflections on X-ray diffraction patterns obtained from the prepared specimens. The test material was cast steel containing (in wt.%: 0.29%C, 1.02%Mn, 4.36%Si, 0.007%S, 0.015%P, 17.8%Cr, 29.3%Ni, 1.59%Nb and 1.19%Ti, subjected to the process of annealing at a temperature of 850oC for 100 hours. For identification of structural constituents by the technique of X-ray phase analysis, four solid specimens were prepared. Their surfaces were successively ground, polished, and subjected to electrolytic etching. The reference sample (isolate was obtained by the method of electrolytic extraction. In solid material the following phases were identified: Feγ, NbC and G; in the isolate additionally the presence of TiC, M23C6 and σ was reported. It has been proved that in the case of solid specimens partial identification of phase constituents may be carried out on surfaces subjected only to grinding with 600 grit abrasive paper without the need of any further preparation.

  8. The new sample preparation line for radiocarbon measurements on atmospheric aerosol at LABEC

    Energy Technology Data Exchange (ETDEWEB)

    Calzolai, G., E-mail: calzolai@fi.infn.i [Department of Physics and Astronomy, Universita degli Studi di Firenze, Florence (Italy); INFN - Istituto Nazionale di Fisica Nucleare, Florence (Italy); Bernardoni, V. [Department of Physics, Universita degli Studi di Milano and INFN, Milan (Italy); Chiari, M.; Fedi, M.E. [INFN - Istituto Nazionale di Fisica Nucleare, Florence (Italy); Lucarelli, F. [Department of Physics and Astronomy, Universita degli Studi di Firenze, Florence (Italy); INFN - Istituto Nazionale di Fisica Nucleare, Florence (Italy); Nava, S. [INFN -Istituto Nazionale di Fisica Nucleare, Florence (Italy); Riccobono, F. [Department of Physics, Universita degli Studi di Milano and INFN, Milan (Italy); Taccetti, F. [INFN - Istituto Nazionale di Fisica Nucleare, Florence (Italy); Valli, G.; Vecchi, R. [Department of Physics, Universita degli Studi di Milano and INFN, Milan (Italy)


    Research highlights: {yields} A new sample preparation line for {sup 14}C analysis on aerosol samples was set up at LABEC. {yields} Effective combustion, selection and collection of the CO{sub 2} were achieved. {yields} The efficiency of the line is consistent with 100%. {yields} AMS tests of samples: reproducibility better than 3 per mille ; good background and accuracy.The line is designed to allow future {sup 14}C measurements on OC/EC. -- Abstract: Radiocarbon measurements on the carbonaceous aerosol fractions have been demonstrated as an effective tool for aerosol source apportionment. For these measurements, a new sample preparation facility was installed at the INFN-LABEC laboratory of Florence (Italy). The line was designed to allow the preparation of samples from different carbonaceous fractions: the combustion of the aerosol samples can be performed in helium or oxygen flows, according to thermal sequences. The evolved CO{sub 2} is cryogenically trapped and reduced to graphite, which is the target material for following Accelerator Mass Spectrometry (AMS) {sup 14}C measurements. This preparation line is described in detail in the paper. As a first step, the line was tested by means of AMS measurements performed on standards to check the reproducibility and the accuracy of the system; moreover, preliminary measurements on the total carbon fraction in aerosol samples were made. Results of these measurements are also reported.

  9. Liquid-Based Medium Used to Prepare Cytological Breast Nipple Fluid Improves the Quality of Cellular Samples Automatic Collection (United States)

    Zonta, Marco Antonio; Velame, Fernanda; Gema, Samara; Filassi, Jose Roberto; Longatto-Filho, Adhemar


    Background Breast cancer is the second cause of death in women worldwide. The spontaneous breast nipple discharge may contain cells that can be analyzed for malignancy. Halo® Mamo Cyto Test (HMCT) was recently developed as an automated system indicated to aspirate cells from the breast ducts. The objective of this study was to standardize the methodology of sampling and sample preparation of nipple discharge obtained by the automated method Halo breast test and perform cytological evaluation in samples preserved in liquid medium (SurePath™). Methods We analyzed 564 nipple fluid samples, from women between 20 and 85 years old, without history of breast disease and neoplasia, no pregnancy, and without gynecologic medical history, collected by HMCT method and preserved in two different vials with solutions for transport. Results From 306 nipple fluid samples from method 1, 199 (65%) were classified as unsatisfactory (class 0), 104 (34%) samples were classified as benign findings (class II), and three (1%) were classified as undetermined to neoplastic cells (class III). From 258 samples analyzed in method 2, 127 (49%) were classified as class 0, 124 (48%) were classified as class II, and seven (2%) were classified as class III. Conclusion Our study suggests an improvement in the quality and quantity of cellular samples when the association of the two methodologies is performed, Halo breast test and the method in liquid medium. PMID:29147397

  10. Detection of Acetone Processing of Castor Bean Mash for Forensic Investigation of Ricin Preparation Methods

    Energy Technology Data Exchange (ETDEWEB)

    Kreuzer-Martin, Helen W.; Wahl, Jon H.; Metoyer, Candace N.; Colburn, Heather A.; Wahl, Karen L.


    The toxic protein ricin is of concern as a potential biological threat agent (BTA) Recently, several samples of ricin have been seized in connection with biocriminal activity. Analytical methods are needed that enable federal investigators to determine how the samples were prepared, to match seized samples to potential source materials, and to identify samples that may have been prepared by the same method using the same source materials. One commonly described crude ricin preparation method is acetone extraction of crushed castor beans. Here we describe the use of solid-phase microextraction and headspace analysis of crude ricin preparation samples to determine whether they were processed by acetone extraction. In all cases, acetone-extracted bean mash could be distinguished from un-extracted mash or mash extracted with other organic solvents. Statistical analysis showed that storage in closed containers for up to 109 days had no effect on acetone signal intensity. Signal intensity in acetone-extracted mash decreased during storage in open containers, but extracted mash could still be distinguished from un-extracted mash after 94 days.

  11. Applications and Preparation Methods of Copper Chromite Catalysts: A Review

    Directory of Open Access Journals (Sweden)

    Ram Prasad


    Full Text Available In this review article various applications and preparation methods of copper chromite catalysts have been discussed. While discussing it is concluded that copper chromite is a versatile catalyst which not only catalyses numerous processes of commercial importance and national program related to defence and space research but also finds applications in the most concerned problem worldwide i.e. environmental pollution control. Several other very useful applications of copper chromite catalysts are in production of clean energy, drugs and agro chemicals, etc. Various preparation methods about 15 have been discussed which depicts clear idea about the dependence of catalytic activity and selectivity on way of preparation of catalyst. In view of the globally increasing interest towards copper chromite catalysis, reexamination on the important applications of such catalysts and their useful preparation methods is thus the need of the time. This review paper encloses 369 references including a well-conceivable tabulation of the newer state of the art. Copyright © 2011 by BCREC UNDIP. All rights reserved.(Received: 19th March 2011, Revised: 03rd May 2011, Accepted: 23rd May 2011[How to Cite: R. Prasad, and P. Singh. (2011. Applications and Preparation Methods of Copper Chromite Catalysts: A Review. Bulletin of Chemical Reaction Engineering & Catalysis, 6 (2: 63-113. doi:10.9767/bcrec.6.2.829.63-113][How to Link / DOI: || or local: ] | View in 

  12. Large sample neutron activation analysis avoids representative sub-sampling and sample preparation difficulties : An added value for forensic analysis

    NARCIS (Netherlands)

    Bode, P.; Romanò, Sabrina; Romolo, Francesco Saverio


    A crucial part of any chemical analysis is the degree of representativeness of the measurand(s) in the test portion for the same measurands in the object, originally collected for investigation. Such an object usually may have either to be homogenized and sub-sampled, or digested/dissolved. Any

  13. Diagnostic PCR: validation and sample preparation are two sides of the same coin

    DEFF Research Database (Denmark)

    Hoorfar, Jeffrey; Wolffs, Petra; Radstrøm, Peter


    Increased use of powerful PCR technology for the routine detection of pathogens has focused attention on the need for international validation and preparation of official non-commercial guidelines. Bacteria of epidemiological importance should be the prime focus, although a "validation infrastruc...... of quantitative reference DNA material and reagents, production of stringent protocols and tools for thermal cycler performance testing, uncomplicated sample preparation techniques, and extensive ring trials for assessment of the efficacy of selected matrix/pathogen detection protocols....

  14. Method for preparing metal powder, device for preparing metal powder, method for processing spent nuclear fuel (United States)

    Park, Jong-Hee [Clarendon Hills, IL


    A method for producing metal powder is provided the comprising supplying a molten bath containing a reducing agent, contacting a metal oxide with the molten bath for a time and at a temperature sufficient to reduce the metal in the metal oxide to elemental metal and produce free oxygen; and isolating the elemental metal from the molten bath.

  15. Determination of protein carbonyls in plasma, cell extracts, tissue homogenates, isolated proteins: Focus on sample preparation and derivatization conditions

    Directory of Open Access Journals (Sweden)

    Daniela Weber


    Full Text Available Protein oxidation is involved in regulatory physiological events as well as in damage to tissues and is thought to play a key role in the pathophysiology of diseases and in the aging process. Protein-bound carbonyls represent a marker of global protein oxidation, as they are generated by multiple different reactive oxygen species in blood, tissues and cells. Sample preparation and stabilization are key steps in the accurate quantification of oxidation-related products and examination of physiological/pathological processes. This review therefore focuses on the sample preparation processes used in the most relevant methods to detect protein carbonyls after derivatization with 2,4-dinitrophenylhydrazine with an emphasis on measurement in plasma, cells, organ homogenates, isolated proteins and organelles. Sample preparation, derivatization conditions and protein handling are presented for the spectrophotometric and HPLC method as well as for immunoblotting and ELISA. An extensive overview covering these methods in previously published articles is given for researchers who plan to measure protein carbonyls in different samples.

  16. Determination of protein carbonyls in plasma, cell extracts, tissue homogenates, isolated proteins: Focus on sample preparation and derivatization conditions. (United States)

    Weber, Daniela; Davies, Michael J; Grune, Tilman


    Protein oxidation is involved in regulatory physiological events as well as in damage to tissues and is thought to play a key role in the pathophysiology of diseases and in the aging process. Protein-bound carbonyls represent a marker of global protein oxidation, as they are generated by multiple different reactive oxygen species in blood, tissues and cells. Sample preparation and stabilization are key steps in the accurate quantification of oxidation-related products and examination of physiological/pathological processes. This review therefore focuses on the sample preparation processes used in the most relevant methods to detect protein carbonyls after derivatization with 2,4-dinitrophenylhydrazine with an emphasis on measurement in plasma, cells, organ homogenates, isolated proteins and organelles. Sample preparation, derivatization conditions and protein handling are presented for the spectrophotometric and HPLC method as well as for immunoblotting and ELISA. An extensive overview covering these methods in previously published articles is given for researchers who plan to measure protein carbonyls in different samples. © 2015 Published by Elsevier Ltd.

  17. Determination of Cd and Pb in food slurries by GFAAS using cryogenic grinding for sample preparation. (United States)

    Santos, D; Barbosa, F; Tomazelli, A C; Krug, F J; Nóbrega, J A; Arruda, M A Z


    A simple method combining slurry sampling after cryogenic grinding and the use of a permanent modification of the integrated platform inside the transversely heated graphite atomizer (THGA) was proposed for the determination of Cd and Pb in foods. Potentialities of the cryogenic grinding were evaluated for grinding different materials of difficult homogenization such as high fat and high fiber tissues. Animal and vegetal samples were cut into small pieces and ground in liquid nitrogen for 2 min. Slurries were prepared directly in the autosampler cup after cryogenic grinding by transferring an exact amount of homogeneous powdered material (5-20 mg) to the cup, followed by 1.00 mL of 0.2% (v/v) HNO3 containing 0.04% (v/v) Triton X-100 and sonication for 30 s, before transferring into the platform previously coated with 250 microg W and 200 microg Rh. Use of a tungsten carbide-rhodium permanent modifier combined with NH4H2PO4 conventional modifier improves tube lifetime and increases the pyrolysis temperature for Cd. Homogeneity tests, carried out by comparing the between- and within-batch precision for each kind of sample, showed no significant differences at the 95% confidence level, indicating good homogeneity for 5-20 mg masses. Detection limits were 3.3 ng g(-1) Cd and 75 ng g(-1) Pb for 1% m/v slurries. Results for determination of Cd and Pb in foods slurries were in agreement with those obtained with digested samples, since no statistical differences were found by the paired t-test at the 95% level.

  18. Some connections between importance sampling and enhanced sampling methods in molecular dynamics (United States)

    Lie, H. C.; Quer, J.


    In molecular dynamics, enhanced sampling methods enable the collection of better statistics of rare events from a reference or target distribution. We show that a large class of these methods is based on the idea of importance sampling from mathematical statistics. We illustrate this connection by comparing the Hartmann-Schütte method for rare event simulation (J. Stat. Mech. Theor. Exp. 2012, P11004) and the Valsson-Parrinello method of variationally enhanced sampling [Phys. Rev. Lett. 113, 090601 (2014)]. We use this connection in order to discuss how recent results from the Monte Carlo methods literature can guide the development of enhanced sampling methods.

  19. Comparative Evaluation of Physicochemical Properties of Pine Needle Powders Prepared by Different Drying Methods


    Chung, Ha-Sook; Lee, Jun Ho


    Systematic study of how different drying methods, namely hot-air drying, vacuum-drying, and freeze-drying, affect color, browning index, degree of rehydration, water solubility, and vitamin C content is critical for utilizing pine needle powders (PNP) as a novel ingredient in functional foods. Samples prepared by vacuum-drying showed a significantly higher L*-value, whereas higher a*- and b*-values were detected in the hot-air dried samples (P

  20. Analysis of acute myelogenous leukemia: preparation of samples for genomic and proteomic analyses. (United States)

    Gjertsen, Bjørn Tore; Øyan, Anne M; Marzolf, Bruz; Hovland, Randi; Gausdal, Gro; Døskeland, Stein-Ove; Dimitrov, Krassen; Golden, Allison; Kalland, Karl-Henning; Hood, Leroy; Bruserud, Øystein


    During the last decade, several large clinical studies have demonstrated that analysis of chromosomal abnormalities is an essential basis for therapeutic decisions in patients with acute myelogenous leukemia (AML), and cytogenetic studies should now be regarded as mandatory both for routine treatment and as a part of clinical investigations in AML. However, new techniques for detailed genetic characterization and analysis of gene expression as well as protein modulation will become important in the further classification of AML subsets and the development of risk-adapted therapeutic strategies. In this context, we emphasize the importance of population-based clinical studies as a basis for future therapeutic guidelines. Such studies will then require the inclusion of patients at small clinical centers without specialized hematological research laboratories. To document a high and uniform quality of the laboratory investigations, it will be necessary to collect material for later analysis in selected laboratories. In this article, we describe current methods for collection of biological samples that can be used for later preparation of DNA, RNA, and proteins. With the use of gradient-separated AML cells, it should be possible to establish the necessary techniques for collection and handling of biological samples even at smaller centers, and complete collections from all included patients should then be possible even in population-based clinical studies.

  1. Preparation of clay mineral samples for high resolution x-ray imaging (United States)

    Abbati, Gennaro; Seim, Christian; Legall, Herbert; Stiel, Holger; Thomas, Noel; Wilhein, Thomas


    In the development of optimum ceramic materials for plastic forming, it is of fundamental importance to gain insight into the compositions of the clay minerals. Whereas spectroscopic methods are adequate for determining the elemental composition of a given sample, a knowledge of the spatial composition, together with the shape and size of the particles leads to further, valuable insight. This requires an imaging technique such as high resolution X-ray microscopy. In addition, fluorescence spectroscopy provides a viable element mapping technique. Since the fine particle fraction of the materials has a major effect on physical properties like plasticity, the analysis is focused mainly on the smallest particles. To separate these from the bigger agglomerates, the raw material has to pass through several procedures like centrifugation and filtering. After that, one has to deposit a layer of appropriate thickness on to a suitable substrate. These preparative techniques are described here, starting from the clay mineral raw materials and proceeding through to samples that are ready to analyze. First results using high resolution x-ray imaging are shown.

  2. 19 CFR 151.83 - Method of sampling. (United States)


    ... 19 Customs Duties 2 2010-04-01 2010-04-01 false Method of sampling. 151.83 Section 151.83 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY (CONTINUED) EXAMINATION, SAMPLING, AND TESTING OF MERCHANDISE Cotton § 151.83 Method of sampling. For...

  3. 7 CFR 29.110 - Method of sampling. (United States)


    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Method of sampling. 29.110 Section 29.110 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing... INSPECTION Regulations Inspectors, Samplers, and Weighers § 29.110 Method of sampling. In sampling tobacco...

  4. an assessment of methods for sampling carabid beetles

    African Journals Online (AJOL)


    particular habitat where we sampled (rugged montane rain forest) pitfall trapping has no advantage over searching methods with respect to ease of operation, low cost or efficiency. However, despite its inefficiency, pitfall trapping cannot be left out of sampling protocols because the method sampled some species that were ...

  5. Sampling Methods in Cardiovascular Nursing Research: An Overview. (United States)

    Kandola, Damanpreet; Banner, Davina; O'Keefe-McCarthy, Sheila; Jassal, Debbie


    Cardiovascular nursing research covers a wide array of topics from health services to psychosocial patient experiences. The selection of specific participant samples is an important part of the research design and process. The sampling strategy employed is of utmost importance to ensure that a representative sample of participants is chosen. There are two main categories of sampling methods: probability and non-probability. Probability sampling is the random selection of elements from the population, where each element of the population has an equal and independent chance of being included in the sample. There are five main types of probability sampling including simple random sampling, systematic sampling, stratified sampling, cluster sampling, and multi-stage sampling. Non-probability sampling methods are those in which elements are chosen through non-random methods for inclusion into the research study and include convenience sampling, purposive sampling, and snowball sampling. Each approach offers distinct advantages and disadvantages and must be considered critically. In this research column, we provide an introduction to these key sampling techniques and draw on examples from the cardiovascular research. Understanding the differences in sampling techniques may aid nurses in effective appraisal of research literature and provide a reference pointfor nurses who engage in cardiovascular research.

  6. Method of preparing nuclear wastes for tansportation and interim storage (United States)

    Bandyopadhyay, Gautam; Galvin, Thomas M.


    Nuclear waste is formed into a substantially water-insoluble solid for temporary storage and transportation by mixing the calcined waste with at least 10 weight percent powdered anhydrous sodium silicate to form a mixture and subjecting the mixture to a high humidity environment for a period of time sufficient to form cementitious bonds by chemical reaction. The method is suitable for preparing an interim waste form from dried high level radioactive wastes.

  7. Methods used to streamline data preparation for memory products (United States)

    DePesa, Paul; Leitermann, Wolfgang


    Historically the transcription of design data, in preparation for mask manufacture, has been to produce flat exposure tool data. Recent developments in hierarchical fracturing in the CATS' tool are reviewed and explained. Current design data, especially memory products or even microprocessors containing cache, benefit immensely from these developments. The pertinent CATS' commands are reviewed. Records of typical data fractures are presented and reviewed, showing the overall decrease in CPU usage with the hierarchal methods. The concomitant decrease in file sizes is also shown.

  8. Preparing Silica Aerogel Monoliths via a Rapid Supercritical Extraction Method (United States)

    Gorka, Caroline A.


    A procedure for the fabrication of monolithic silica aerogels in eight hours or less via a rapid supercritical extraction process is described. The procedure requires 15-20 min of preparation time, during which a liquid precursor mixture is prepared and poured into wells of a metal mold that is placed between the platens of a hydraulic hot press, followed by several hours of processing within the hot press. The precursor solution consists of a 1.0:12.0:3.6:3.5 x 10-3 molar ratio of tetramethylorthosilicate (TMOS):methanol:water:ammonia. In each well of the mold, a porous silica sol-gel matrix forms. As the temperature of the mold and its contents is increased, the pressure within the mold rises. After the temperature/pressure conditions surpass the supercritical point for the solvent within the pores of the matrix (in this case, a methanol/water mixture), the supercritical fluid is released, and monolithic aerogel remains within the wells of the mold. With the mold used in this procedure, cylindrical monoliths of 2.2 cm diameter and 1.9 cm height are produced. Aerogels formed by this rapid method have comparable properties (low bulk and skeletal density, high surface area, mesoporous morphology) to those prepared by other methods that involve either additional reaction steps or solvent extractions (lengthier processes that generate more chemical waste).The rapid supercritical extraction method can also be applied to the fabrication of aerogels based on other precursor recipes. PMID:24637334

  9. On Angular Sampling Methods for 3-D Spatial Channel Models

    DEFF Research Database (Denmark)

    Fan, Wei; Jämsä, Tommi; Nielsen, Jesper Ødum


    This paper discusses generating three dimensional (3D) spatial channel models with emphasis on the angular sampling methods. Three angular sampling methods, i.e. modified uniform power sampling, modified uniform angular sampling, and random pairing methods are proposed and investigated in detail....... The random pairing method, which uses only twenty sinusoids in the ray-based model for generating the channels, presents good results if the spatial channel cluster is with a small elevation angle spread. For spatial clusters with large elevation angle spreads, however, the random pairing method would fail...

  10. Photocatalytic degradation of textile dyestuffs using TiO{sub 2} nanotubes prepared by sonoelectrochemical method

    Energy Technology Data Exchange (ETDEWEB)

    Tekin, Derya, E-mail:


    Highlights: • TiO{sub 2} nanotubes prepared by electrochemical and sonoelectrochemical method. • More regular TiO{sub 2} nanotubes diameters prepared by sonoelectrochemical method. • Obtained nanotubes were used in the photocatalytic degradation of Orange G dye. • TiO{sub 2} nanotubes prepared by sonoelectrochemical method showed 10% faster degradation of Orange G dye compared with the one by electrochemical method. - Abstract: TiO{sub 2} nanotubes were prepared by anodization of Ti plates by conventional electrochemical technique as well as by an emerging sonoelectrochemical technique. Scanning electron miscroscope (SEM) analysis showed that ultrasound assisted anodization yielded more ordered and controllable TiO{sub 2} tube banks with higher tube diameter. The photocatalytical activities of TiO{sub 2} nanotubes were tested in the photocatalytical degradation of Orange G dye. The results showed that sonoelectrochemically prepared TiO{sub 2} tubes exhibited 10% higher photocatalytic performance than the electrochemical prepared ones, and more than 18% higher activity than the other TiO{sub 2} samples.

  11. A comparative study of ChIP-seq sequencing library preparation methods. (United States)

    Sundaram, Arvind Y M; Hughes, Timothy; Biondi, Shea; Bolduc, Nathalie; Bowman, Sarah K; Camilli, Andrew; Chew, Yap C; Couture, Catherine; Farmer, Andrew; Jerome, John P; Lazinski, David W; McUsic, Andrew; Peng, Xu; Shazand, Kamran; Xu, Feng; Lyle, Robert; Gilfillan, Gregor D


    ChIP-seq is the primary technique used to investigate genome-wide protein-DNA interactions. As part of this procedure, immunoprecipitated DNA must undergo "library preparation" to enable subsequent high-throughput sequencing. To facilitate the analysis of biopsy samples and rare cell populations, there has been a recent proliferation of methods allowing sequencing library preparation from low-input DNA amounts. However, little information exists on the relative merits, performance, comparability and biases inherent to these procedures. Notably, recently developed single-cell ChIP procedures employing microfluidics must also employ library preparation reagents to allow downstream sequencing. In this study, seven methods designed for low-input DNA/ChIP-seq sample preparation (Accel-NGS® 2S, Bowman-method, HTML-PCR, SeqPlex™, DNA SMART™, TELP and ThruPLEX®) were performed on five replicates of 1 ng and 0.1 ng input H3K4me3 ChIP material, and compared to a "gold standard" reference PCR-free dataset. The performance of each method was examined for the prevalence of unmappable reads, amplification-derived duplicate reads, reproducibility, and for the sensitivity and specificity of peak calling. We identified consistent high performance in a subset of the tested reagents, which should aid researchers in choosing the most appropriate reagents for their studies. Furthermore, we expect this work to drive future advances by identifying and encouraging use of the most promising methods and reagents. The results may also aid judgements on how comparable are existing datasets that have been prepared with different sample library preparation reagents.

  12. A sample preparation process for LC-MS/MS analysis of total protein drug concentrations in monkey plasma samples with antibody. (United States)

    Ji, Qin C; Rodila, Ramona; El-Shourbagy, Tawakol A


    The determination of protein concentrations in plasma samples often provides essential information in biomedical research, clinical diagnostics, and pharmaceutical discovery and development. Binding assays such as ELISA determine meaningful free analyte concentrations by using specific antigen or antibody reagents. Concurrently, mass spectrometric technology is becoming a promising complementary method to traditional binding assays. Mass spectrometric assays generally provide measurements of the total protein analyte concentration. However, it was found that antibodies may bind strongly with the protein analyte such that total concentrations cannot be determined. Thus, a sample preparation process was developed which included a novel "denaturing" step to dissociate binding between antibodies and the protein analyte prior to solid phase extraction of plasma samples and LC-MS/MS analysis. In so doing, the total protein analyte concentrations can be obtained. This sample preparation process was further studied by LC-MS analysis with a full mass range scan. It was found that the protein of interest and other plasma peptides were pre-concentrated, while plasma albumin was depleted in the extracts. This capability of the sample preparation process could provide additional advantages in proteomic research for biomarker discovery and validation. The performance of the assay with the novel denaturing step was further evaluated. The linear dynamic range was between 100.9ng/mL and 53920.0ng/mL with a coefficient of determination (r(2)) ranging from 0.9979 and 0.9997. For LLOQ and ULOQ samples, the inter-assay CV was 12.6% and 2.7% and inter-assay mean accuracies were 103.7% and 99.5% of theoretical concentrations, respectively. For QC samples, the inter-assay CV was between 2.1% and 4.9%, and inter-assay mean accuracies were between 104.1% and 110.0% of theoretical concentrations.

  13. Separation of four flavonoids from Rhodiola rosea by on-line combination of sample preparation and counter-current chromatography. (United States)

    Ma, Chaoyang; Hu, Liming; Fu, Qianyun; Gu, Xiaohong; Tao, Guanjun; Wang, Hongxin


    Purification of four flavonoids from Rhodiola rosea was developed by on-line combination of sample preparation and counter-current chromatography (CCC). Flavonoid sample was prepared by dynamic ultrasonic-assisted and solid-phase extraction using ion liquids as extractant. The preparation conditions were optimized by D-optimal design as follows: 2mol/L of 1-ethyl-3-methylimidazolium bromide concentration, 360W of ultrasonic power, 1.5mL/min of flow rate, 35min of extraction time and 0.5mL (absorbent) per g (material) of absorbent amount. The prepared sample solution (20mL) was loaded and injected directly into CCC column for final separation. As a result, four flavonoids, herbacetin-3-O-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside 1 (40.1mg), kaempferol-3-O-β-d-glucopyranosyl-7-O-α-l-rhamn-opyranoside 2 (4.6mg), kaempferol 3-O-β-d-glucopyranoside-(2→1)-β-d-xylopyranoside 3 (20.2mg) and herbacetin-8-O-β-d-glucopyranoside 4 (22.5mg), were obtained from 20g of R. rosea material using ethyl acetate-n-butanol-H2O as solvent system at a ratio of 4:1:5 by CCC. Their structures were identified by ESI-MS/MS, NMR methods. Their purities determined by UPLC were 98.5%, 95.4%, 98.1% and 97.5%, respectively. Kaempferol-3-O-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside 2 and herbacetin-8-O-β-d-glucopyrano-side 4 were isolated for first time from R. rosea. The purification method was simple, efficient and evaded tedious sample preparation process. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. New adaptive sampling method in particle image velocimetry (United States)

    Yu, Kaikai; Xu, Jinglei; Tang, Lan; Mo, Jianwei


    This study proposes a new adaptive method to enable the number of interrogation windows and their positions in a particle image velocimetry (PIV) image interrogation algorithm to become self-adapted according to the seeding density. The proposed method can relax the constraint of uniform sampling rate and uniform window size commonly adopted in the traditional PIV algorithm. In addition, the positions of the sampling points are redistributed on the basis of the spring force generated by the sampling points. The advantages include control of the number of interrogation windows according to the local seeding density and smoother distribution of sampling points. The reliability of the adaptive sampling method is illustrated by processing synthetic and experimental images. The synthetic example attests to the advantages of the sampling method. Compared with that of the uniform interrogation technique in the experimental application, the spatial resolution is locally enhanced when using the proposed sampling method.

  15. A New Automated Method and Sample Data Flow for Analysis of Volatile Nitrosamines in Human Urine. (United States)

    Hodgson, James A; Seyler, Tiffany H; McGahee, Ernest; Arnstein, Stephen; Wang, Lanqing


    Volatile nitrosamines (VNAs) are a group of compounds classified as probable (group 2A) and possible (group 2B) carcinogens in humans. Along with certain foods and contaminated drinking water, VNAs are detected at high levels in tobacco products and in both mainstream and sidestream smoke. Our laboratory monitors six urinary VNAs-N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosodiethylamine (NDEA), N-nitrosopiperidine (NPIP), N-nitrosopyrrolidine (NPYR), and N-nitrosomorpholine (NMOR)-using isotope dilution GC-MS/MS (QQQ) for large population studies such as the National Health and Nutrition Examination Survey (NHANES). In this paper, we report for the first time a new automated sample preparation method to more efficiently quantitate these VNAs. Automation is done using Hamilton STAR™ and Caliper Staccato™ workstations. This new automated method reduces sample preparation time from 4 hours to 2.5 hours while maintaining precision (inter-run CV method increases sample throughput while maintaining a low limit of detection (sample data flow was created in parallel to the automated method, in which samples can be tracked from receiving to final LIMs output with minimal human intervention, further minimizing human error in the sample preparation process. This new automated method and the sample data flow are currently applied in bio-monitoring of VNAs in the US non-institutionalized population NHANES 2013-2014 cycle.

  16. Preparation of magnetic graphene @polydopamine @Zr-MOF material for the extraction and analysis of bisphenols in water samples. (United States)

    Wang, Xianying; Deng, Chunhui


    In this work, a simple method for the extraction and analysis of bisphenols in environmental samples was presented. And the prepared zirconium-based magnetic MOFs (magG@PDA@Zr-MOF) were used as the sorbents for the magnetic solid-phase extraction. With the simple solvothermal reaction and sol-gel method, the prepared material showed great characteristics of large surface area, homogeneous pore size, good magnetic responsivity and super-hydrophilicity. The large surface area provided abundant sites to extract target compounds; the magnetic property could simplify the whole extraction procedure; and the hydrophilicity improved the dispersibility of the material in matrix. Here, various extraction parameters were optimized, including amounts of sorbents, adsorption time, species of elution solvents and desorption time. The whole extraction procedure could be accomplished in 30 min. And under the optimized conditions, method validations were also studied, such as linearity, the limit of detection and recovery. Finally, the prepared material was used in real water samples. The results showed this material had good potential as the sorbent for the extraction of targets in environmental water samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Author Contribution to the Pu Handbook II: Chapter 37 LLNL Integrated Sample Preparation Glovebox (TEM) Section

    Energy Technology Data Exchange (ETDEWEB)

    Wall, Mark A. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)


    The development of our Integrated Actinide Sample Preparation Laboratory (IASPL) commenced in 1998 driven by the need to perform transmission electron microscopy studies on naturally aged plutonium and its alloys looking for the microstructural effects of the radiological decay process (1). Remodeling and construction of a laboratory within the Chemistry and Materials Science Directorate facilities at LLNL was required to turn a standard radiological laboratory into a Radiological Materials Area (RMA) and Radiological Buffer Area (RBA) containing type I, II and III workplaces. Two inert atmosphere dry-train glove boxes with antechambers and entry/exit fumehoods (Figure 1), having a baseline atmosphere of 1 ppm oxygen and 1 ppm water vapor, a utility fumehood and a portable, and a third double-walled enclosure have been installed and commissioned. These capabilities, along with highly trained technical staff, facilitate the safe operation of sample preparation processes and instrumentation, and sample handling while minimizing oxidation or corrosion of the plutonium. In addition, we are currently developing the capability to safely transfer small metallographically prepared samples to a mini-SEM for microstructural imaging and chemical analysis. The gloveboxes continue to be the most crucial element of the laboratory allowing nearly oxide-free sample preparation for a wide variety of LLNL-based characterization experiments, which includes transmission electron microscopy, electron energy loss spectroscopy, optical microscopy, electrical resistivity, ion implantation, X-ray diffraction and absorption, magnetometry, metrological surface measurements, high-pressure diamond anvil cell equation-of-state, phonon dispersion measurements, X-ray absorption and emission spectroscopy, and differential scanning calorimetry. The sample preparation and materials processing capabilities in the IASPL have also facilitated experimentation at world-class facilities such as the

  18. Bovine liver sample preparation and micro-homogeneity study for Cu and Zn determination by solid sampling electrothermal atomic absorption spectrometry (United States)

    Nomura, Cassiana S.; Silva, Cíntia S.; Nogueira, Ana R. A.; Oliveira, Pedro V.


    This work describes a systematic study for the bovine liver sample preparation for Cu and Zn determination by solid sampling electrothermal atomic absorption spectrometry. The main parameters investigated were sample drying, grinding process, particle size, sample size, microsample homogeneity, and their relationship with the precision and accuracy of the method. A bovine liver sample was prepared using different drying procedures: (1) freeze drying, and (2) drying in a household microwave oven followed by drying in a stove at 60 °C until constant mass. Ball and cryogenic mills were used for grinding. Less sensitive wavelengths for Cu (216.5 nm) and Zn (307.6 nm), and Zeeman-based three-field background correction for Cu were used to diminish the sensitivities. The pyrolysis and atomization temperatures adopted were 1000 °C and 2300 °C for Cu, and 700 °C and 1700 °C for Zn, respectively. For both elements, it was possible to calibrate the spectrometer with aqueous solutions. The use of 250 μg of W + 200 μg of Rh as permanent chemical modifier was imperative for Zn. Under these conditions, the characteristic mass and detection limit were 1.4 ng and 1.6 ng for Cu, and 2.8 ng and 1.3 ng for Zn, respectively. The results showed good agreement (95% confidence level) for homogeneity of the entire material (> 200 mg) when the sample was dried in microwave/stove and ground in a cryogenic mill. The microsample homogeneity study showed that Zn is more dependent on the sample pretreatment than Cu. The bovine liver sample prepared in microwave/stove and ground in a cryogenic mill presented results with the lowest relative standard deviation for Cu than Zn. Good accuracy and precision were observed for bovine liver masses higher than 40 μg for Cu and 30 μg for Zn. The concentrations of Cu and Zn in the prepared bovine liver sample were 223 mg kg - 1 and 128 mg kg - 1, respectively. The relative standard deviations were lower than 6% ( n = 5). The accuracy of the entire

  19. Preparing to Receive and Handle Martian Samples When They Arrive on Earth (United States)

    McCubbin, Francis M.


    The Astromaterials Acquisition and Curation Office at NASA Johnson Space Center (JSC) is responsible for curating all of NASA's extraterrestrial samples. Under the governing document, NASA Policy Directive (NPD) 7100.10F+ derivative NPR 'Curation of Extraterrestrial Materials', JSC is charged with 'The curation of all extraterrestrial material under NASA control, including future NASA missions. 'The Directive goes on to define Curation as including'...documentation, preservation, preparation, and distribution of samples for research, education, and public outreach."

  20. A Loop-Mediated Isothermal Amplification Assay and Sample Preparation Procedure for Sensitive Detection of Xanthomonas fragariae in Strawberry (United States)

    Wang, Hehe; Turechek, William W.


    Xanthomonas fragariae is a bacterium that causes angular leaf spot of strawberry. Asymptomatic infection is common and contributes to the difficulties in disease management. The aim of this study was to develop a loop-mediated isothermal amplification (LAMP) assay as an efficient method for detection of asymptomatic infections of X. fragariae. In addition, a new method of sample preparation was developed that allows sampling of a larger amount of plant tissue, hence increasing the detection rate in real-life samples. The sample preparation procedure includes an overnight incubation of strawberry tissues in phosphate-buffered saline (PBS), followed by a quick sample concentration and a boiling step to extract DNA for amplification. The detection limit of the LAMP assay was approximately 2×103 CFU/mL for pure bacteria culture and 300 CFU/mL for bacteria spiked strawberry leaf and petiole samples. LAMP provided a 2–3 fold lower detection limit than the standard qPCR assay but was faster, and more user-friendly. The LAMP assay should serve as a rapid, sensitive and cost-effective tool for detecting asymptomatic infections of X. fragariae in strawberry nursery stock and contribute to improved disease management. PMID:26766068

  1. Soil and Water – What is Detectable through Microbiological Sample Preparation Techniques (United States)

    The concerns of a potential terrorist’s use of biological agents in soil and ground water are articulated by comparisons to major illnesses in this Country involving contaminated drinking water sources. Objectives are focused on the importance of sample preparation in the rapid, ...

  2. Facile method to prepare a transparent superhydrophobic PET film (United States)

    Wang, Jian; Chen, Hong; Wang, Xian; Yuan, Zhiqing


    A simple method to fabricate a transparent superhydrophobic coating was developed by spraying PDMS/SiO2 nanoparticles suspension on a hydrophilic PET film. This PDMS/SiO2 nanoparticles suspension was prepared by fully diluting and properly stirring. When this suspension was sprayed on the surface of PET film at a distance of about 30 cm for 2 s, a porous micro-nano-binary structure was formed, which endowed the hydrophilic PET film superhydrophobicity, while the PET film maintained transparent after spraying with a SiO2 nanoparticle layer. The as-prepared superhydrophobic PET film exhibited a high water contact angle of about 155° ± 1.9° and a low water sliding angle of about 7°. The formation mechanism of this transparent superhydrophobic coating on PET film was also discussed in detail in this paper.

  3. Novel Modification of HistoGel-Based Cell Block Preparation Method: Improved Sufficiency for Molecular Studies. (United States)

    Rekhtman, Natasha; Buonocore, Darren J; Rudomina, Dorota; Friedlander, Maria; Dsouza, Crisbane; Aggarwal, Gitika; Arcila, Maria; Edelweiss, Marcia; Lin, Oscar


    - Cell block preparation methods vary substantially across institutions and are frequently suboptimal. The growing importance of biomarker testing in the era of targeted therapies makes optimization of cell block preparation critically important. - To develop an improved cell block preparation method. - Ex vivo fine-needle aspirates and scrapes from surgically resected tumors were used to develop an improved HistoGel (Thermo Fisher Scientific, Waltham, Massachusetts)-based cell block preparation method. Cellularity yield with the new versus the standard method was assessed in ex vivo split samples and in consecutive clinical fine-needle aspirates processed before (n = 100) and after (n = 100) the new method was implemented in our laboratory. Sufficiency of cell block material for potential molecular studies was estimated by manual cell quantitation. - The key modification in the new method was pretreatment of the pelleted cells with 95% ethanol before the addition of HistoGel (HistoGel + ethanol method). In addition, we optimized the melting conditions of HistoGel and added a dark, inorganic marker to the cell pellets to highlight the desired level of sectioning during microtomy. Cell blocks from ex vivo split samples showed that the HistoGel + ethanol method yielded, on average, an 8.3-fold (range, 1-20) greater cellularity compared with the standard HistoGel-only method. After the switch from the standard HistoGel method to the modified method in our clinical practice, sufficiency of positive fine-needle aspirates for some molecular studies increased from 72% to 97% (P = .002). - We describe a simple and readily adoptable modification of the HistoGel method, which results in substantial improvement in cell capture in cell blocks, leading to significant increase in sufficiency for potential molecular and other ancillary studies.

  4. Systems and methods for self-synchronized digital sampling (United States)

    Samson, Jr., John R. (Inventor)


    Systems and methods for self-synchronized data sampling are provided. In one embodiment, a system for capturing synchronous data samples is provided. The system includes an analog to digital converter adapted to capture signals from one or more sensors and convert the signals into a stream of digital data samples at a sampling frequency determined by a sampling control signal; and a synchronizer coupled to the analog to digital converter and adapted to receive a rotational frequency signal from a rotating machine, wherein the synchronizer is further adapted to generate the sampling control signal, and wherein the sampling control signal is based on the rotational frequency signal.

  5. A novel PFIB sample preparation protocol for correlative 3D X-ray CNT and FIB-TOF-SIMS tomography

    Energy Technology Data Exchange (ETDEWEB)

    Priebe, Agnieszka, E-mail: [Univ. Grenoble Alpes, F-38000 Grenoble (France); CEA, LETI, MINATEC Campus, F-38054 Grenoble (France); Audoit, Guillaume; Barnes, Jean-Paul [Univ. Grenoble Alpes, F-38000 Grenoble (France); CEA, LETI, MINATEC Campus, F-38054 Grenoble (France)


    We present a novel sample preparation method that allows correlative 3D X-ray Computed Nano-Tomography (CNT) and Focused Ion Beam Time-Of-Flight Secondary Ion Mass Spectrometry (FIB-TOF-SIMS) tomography to be performed on the same sample. In addition, our invention ensures that samples stay unmodified structurally and chemically between the subsequent experiments. The main principle is based on modifying the topography of the X-ray CNT experimental setup before FIB-TOF-SIMS measurements by incorporating a square washer around the sample. This affects the distribution of extraction field lines and therefore influences the trajectories of secondary ions that are now guided more efficiently towards the detector. As the result, secondary ion detection is significantly improved and higher, i.e. statistically better, signals are obtained. - Highlights: • Novel sample preparation for correlative 3D X-ray CNT and FIB-TOF-SIMS is presented. • Two experiments are conducted on exactly the same sample without any modifications. • Introduction of a square washer around the sample leads to increased ion detection.

  6. Sample preparation and liquid chromatography-tandem mass spectrometry for multiple steroids in mammalian and avian circulation.

    Directory of Open Access Journals (Sweden)

    Lee Koren

    Full Text Available Blood samples from wild mammals and birds are often limited in volume, allowing researchers to quantify only one or two steroids from a single sample by immunoassays. In addition, wildlife serum or plasma samples are often lipemic, necessitating stringent sample preparation. Here, we validated sample preparation for simultaneous liquid chromatography--tandem mass spectrometry (LC-MS/MS quantitation of cortisol, corticosterone, 11-deoxycortisol, dehydroepiandrosterone (DHEA, 17β-estradiol, progesterone, 17α-hydroxyprogesterone and testosterone from diverse mammalian (7 species and avian (5 species samples. Using 100 µL of serum or plasma, we quantified (signal-to-noise (S/N ratio ≥ 10 4-7 steroids depending on the species and sample, without derivatization. Steroids were extracted from serum or plasma using automated solid-phase extraction where samples were loaded onto C18 columns, washed with water and hexane, and then eluted with ethyl acetate. Quantitation by LC-MS/MS was done in positive ion, multiple reaction-monitoring (MRM mode with an atmospheric pressure chemical ionization (APCI source and heated nebulizer (500°C. Deuterated steroids served as internal standards and run time was 15 minutes. Extraction recoveries were 87-101% for the 8 analytes, and all intra- and inter-run CVs were ≤ 8.25%. This quantitation method yields good recoveries with variable lipid-content samples, avoids antibody cross-reactivity issues, and delivers results for multiple steroids. Thus, this method can enrich datasets by providing simultaneous quantitation of multiple steroids, and allow researchers to reimagine the hypotheses that could be tested with their volume-limited, lipemic, wildlife samples.

  7. Advancement of Solidification Processing Technology Through Real Time X-Ray Transmission Microscopy: Sample Preparation (United States)

    Stefanescu, D. M.; Curreri, P. A.


    Two types of samples were prepared for the real time X-ray transmission microscopy (XTM) characterization. In the first series directional solidification experiments were carried out to evaluate the critical velocity of engulfment of zirconia particles in the Al and Al-Ni eutectic matrix under ground (l-g) conditions. The particle distribution in the samples was recorded on video before and after the samples were directionally solidified. In the second series samples of the above two type of composites were prepared for directional solidification runs to be carried out on the Advanced Gradient Heating Facility (AGHF) aboard the space shuttle during the LMS mission in June 1996. X-ray microscopy proved to be an invaluable tool for characterizing the particle distribution in the metal matrix samples. This kind of analysis helped in determining accurately the critical velocity of engulfment of ceramic particles by the melt interface in the opaque metal matrix composites. The quality of the cast samples with respect to porosity and instrumented thermocouple sheath breakage or shift could be easily viewed and thus helped in selecting samples for the space shuttle experiments. Summarizing the merits of this technique it can be stated that this technique enabled the use of cast metal matrix composite samples since the particle location was known prior to the experiment.

  8. Optical Properties and Surface Morphology of Zinc Telluride Thin Films Prepared by Stacked Elemental Layer Method

    Directory of Open Access Journals (Sweden)

    Subramani SHANMUGAN


    Full Text Available ZnTe thin films were prepared by Stacking of elemental (Zn and Te layers (SEL followed by inert gas annealing. The optical parameters were calculated from the transmission spectra. The bandgap of the annealed samples was found between 1.95 eV and 2.06 eV. The change in film thickness after annealing was observed using cross sectional SEM image of the annealed samples. The surface morphology of the annealed Te/Zn stack was also analyzed and observed as very smooth, compact and dense surface. The prepared film was Zn rich evidenced by EDAX. The observed result encourages in pursuing the SEL method for the preparation of compound semiconductor from II-VI group materials.DOI:

  9. DOE methods for evaluating environmental and waste management samples.

    Energy Technology Data Exchange (ETDEWEB)

    Goheen, S C; McCulloch, M; Thomas, B L; Riley, R G; Sklarew, D S; Mong, G M; Fadeff, S K [eds.; Pacific Northwest Lab., Richland, WA (United States)


    DOE Methods for Evaluating Environmental and Waste Management Samples (DOE Methods) provides applicable methods in use by. the US Department of Energy (DOE) laboratories for sampling and analyzing constituents of waste and environmental samples. The development of DOE Methods is supported by the Laboratory Management Division (LMD) of the DOE. This document contains chapters and methods that are proposed for use in evaluating components of DOE environmental and waste management samples. DOE Methods is a resource intended to support sampling and analytical activities that will aid in defining the type and breadth of contamination and thus determine the extent of environmental restoration or waste management actions needed, as defined by the DOE, the US Environmental Protection Agency (EPA), or others.

  10. LC-MS analysis of the plasma metabolome–a novel sample preparation strategy

    DEFF Research Database (Denmark)

    Skov, Kasper; Hadrup, Niels; Smedsgaard, Jørn


    of plasma samples: The first is protein precipitation; the second is protein precipitation followed by solid phase extraction with sub-fractionation into three sub-samples; a phospholipid, a lipid and a polar sub-fraction. Molecular feature extraction of the data files from LC-qTOF analysis of the samples......Blood plasma is a well-known body fluid often analyzed in studies on the effects of toxic compounds as physiological or chemical induced changes in the mammalian body are reflected in the plasma metabolome. Sample preparation prior to LC-MS based analysis of the plasma metabolome is a challenge...... revealed 1792 molecular features from the protein precipitation procedure. The protein precipitation followed by solid phase extraction procedure with three sub-samples gave a total of 4234 molecular features. This suggests that sub-sampling into polar, lipid and phospholipid fractions enables extraction...

  11. Comparison of sample preparation procedures on metal(loid) fractionation patterns in lichens. (United States)

    Kroukamp, E M; Godeto, T W; Forbes, P B C


    The effects of different sample preparation strategies and storage on metal(loid) fractionation trends in plant material is largely underresearched. In this study, a bulk sample of lichen Parmotrema austrosinense (Zahlbr.) Hale was analysed for its total extractable metal(loid) content by ICP-MS, and was determined to be adequately homogenous (82% for As, Cu, Mn, Pb, Sr and Zn but poor for other elements, where sample preparation strategies 'no sample preparation' and 'dried in a desiccator' had the best extraction recoveries. Cryogenic freezing procedures had a significantly (p lichens. Biotransformation over a period of a month is suspected for most elements, with the exception of Sr and Zn, where changes in the fractionation patterns were statistically significant (p < 0.05), indicating the need for minimal delay in sample cleaning and preservation when species fractionation patterns are of interest. This study also shows that the assumption that species stability can be ensured through cryopreservation and freeze drying techniques needs to be revisited.

  12. Applied Focused Ion Beam Techniques for Sample Preparation of Astromaterials for Integrated Nano-Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Graham, G A; Teslich, N E; Kearsley, A T; Stadermann, F J; Stroud, R M; Dai, Z R; Ishii, H A; Hutcheon, I D; Bajt, S; Snead, C J; Weber, P K; Bradley, J P


    Sample preparation is always a critical step in study of micrometer sized astromaterials available for study in the laboratory, whether their subsequent analysis is by electron microscopy or secondary ion mass spectrometry. A focused beam of gallium ions has been used to prepare electron transparent sections from an interplanetary dust particle, as part of an integrated analysis protocol to maximize the mineralogical, elemental, isotopic and spectroscopic information extracted from one individual particle. In addition, focused ion beam techniques have been employed to extract cometary residue preserved on the rims and walls of micro-craters in 1100 series aluminum foils that were wrapped around the sample tray assembly on the Stardust cometary sample collector. Non-ideal surface geometries and inconveniently located regions of interest required creative solutions. These include support pillar construction and relocation of a significant portion of sample to access a region of interest. Serial sectioning, in a manner similar to ultramicrotomy, is a significant development and further demonstrates the unique capabilities of focused ion beam microscopy for sample preparation of astromaterials.

  13. An overview of sample preparation and extraction of synthetic pyrethroids from water, sediment and soil. (United States)

    Albaseer, Saeed S; Rao, R Nageswara; Swamy, Y V; Mukkanti, K


    The latest developments in sample preparation and extraction of synthetic pyrethroids from environmental matrices viz., water, sediment and soil were reviewed. Though the synthetic pyrethroids were launched in 1970s, to the best of authors' knowledge there was no review on this subject until date. The present status and recent advances made during the last 10 years in sample preparation including conservation and extraction techniques used in determination of synthetic pyrethroids in water, sediment and soil were discussed. Pre- and post-extraction treatments, sample stability during extraction and its influence upon the whole process of analytical determination were covered. Relative merits and demerits including the green aspects of extraction were evaluated. The current trends and future prospects were also addressed. 2010 Elsevier B.V. All rights reserved.

  14. Automated cellular sample preparation using a Centrifuge-on-a-Chip. (United States)

    Mach, Albert J; Kim, Jae Hyun; Arshi, Armin; Hur, Soojung Claire; Di Carlo, Dino


    The standard centrifuge is a laboratory instrument widely used by biologists and medical technicians for preparing cell samples. Efforts to automate the operations of concentration, cell separation, and solution exchange that a centrifuge performs in a simpler and smaller platform have had limited success. Here, we present a microfluidic chip that replicates the functions of a centrifuge without moving parts or external forces. The device operates using a purely fluid dynamic phenomenon in which cells selectively enter and are maintained in microscale vortices. Continuous and sequential operation allows enrichment of cancer cells from spiked blood samples at the mL min(-1) scale, followed by fluorescent labeling of intra- and extra-cellular antigens on the cells without the need for manual pipetting and washing steps. A versatile centrifuge-analogue may open opportunities in automated, low-cost and high-throughput sample preparation as an alternative to the standard benchtop centrifuge in standardized clinical diagnostics or resource poor settings.

  15. Magnetite nanoparticles prepared by co-precipitation method in different conditions

    Energy Technology Data Exchange (ETDEWEB)

    Aphesteguy, J.C., E-mail: [LAFMACEL-INTECIN, Facultad de Ingeniería, UBA, Paseo Colón 850, C1063EHA Buenos Aires (Argentina); Kurlyandskaya, G.V. [Universidad del País Vasco UPV-EHU, Dept. Electricidad y Electronica, 48940 Leioa (Spain); Ural Federal University, Dept. Magnetism and Magnetic Nanomaterials, 620000 Ekaterinburg (Russian Federation); Celis, J.P. de [National Technology University (UTN), Facultad Regional Avellaneda, Department of Chemistry (Argentina); Safronov, A.P. [Ural Federal University, Dept. Magnetism and Magnetic Nanomaterials, 620000 Ekaterinburg (Russian Federation); Institute of Electrophysics UD RAS, Ekaterinburg 620016 (Russian Federation); Schegoleva, N.N. [Institute of Metal Physics UD RAS, Ekaterinburg 620044 (Russian Federation)


    Magnetic nanoparticles (MNPs) of pure magnetite (Fe{sub 3}O{sub 4}) were prepared in an aqueous solution (sample M−I) and in a water-ethyl alcohol mixture (sample M−II) by the co-precipitation method. The structure and magnetic properties of both samples were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), magnetic (M−H) and microwave measurements (FMR). The mean average particle diameter and particle size distribution was evaluated by the Dynamic Light Scattering (DLS) and Brunauer- Emmett-Teller techniques (BET). The Quantitative chemical analysis of iron was performed by Inductively Coupled Plasma (ICP)- Atomic Emission Spectroscopy (AES) technique. The MNPs prepared in aqueous solution show a higher grain than those prepared in the water-ethyl alcohol mixture. The type of phase structure in both cases can be defined as “defective spinel”. The shape of the majority of M−I MNPs is octahedral. The shape of the majority of M−II MNPs is cubic. The specific surface area of MNPs was as high as 14.4 m{sup 2}/g for M−I sample and 77.8 m{sup 2}/g for sample M–II. The obtained saturation magnetization values of 75 emu/g (M−I) and 68 emu/g (M−II) are consistent with expected values for magnetite MNPs of observed sizes. Ferromagnetic resonance (FMR) measurements confirmed that MNPs of both types are magnetically homogeneous materials. FMR lines' position and line widths can be understood by invoking the local dipolar fields, deviations from sphericity, magnetocrystalline anisotropy and stresses. M−I sample shows sizeable zero field microwave absorption which is absent in the M−II case. The differences in microwave behaviour of M−I and M−II MNPs can be used in the design of microwave radiation absorbing multilayers. - Highlights: • Magnetite nanoparticles were prepared in two different conditions. • Specific surface area of sample prepared in water- ethanol mix is

  16. Publishing nutrition research: a review of sampling, sample size, statistical analysis, and other key elements of manuscript preparation, Part 2. (United States)

    Boushey, Carol J; Harris, Jeffrey; Bruemmer, Barbara; Archer, Sujata L


    Members of the Board of Editors recognize the importance of providing a resource for researchers to insure quality and accuracy of reporting in the Journal. This second monograph of a periodic series focuses on study sample selection, sample size, and common statistical procedures using parametric methods, and the presentation of statistical methods and results. Attention to sample selection and sample size is critical to avoid study bias. When outcome variables adhere to a normal distribution, then parametric procedures can be used for statistical inference. Documentation that clearly outlines the steps used in the research process will advance the science of evidence-based practice in nutrition and dietetics. Real examples from problem sets and published literature are provided, as well as reference to books and online resources.

  17. Adjustable virtual pore-size filter for automated sample preparation using acoustic radiation force

    Energy Technology Data Exchange (ETDEWEB)

    Jung, B; Fisher, K; Ness, K; Rose, K; Mariella, R


    We present a rapid and robust size-based separation method for high throughput microfluidic devices using acoustic radiation force. We developed a finite element modeling tool to predict the two-dimensional acoustic radiation force field perpendicular to the flow direction in microfluidic devices. Here we compare the results from this model with experimental parametric studies including variations of the PZT driving frequencies and voltages as well as various particle sizes and compressidensities. These experimental parametric studies also provide insight into the development of an adjustable 'virtual' pore-size filter as well as optimal operating conditions for various microparticle sizes. We demonstrated the separation of Saccharomyces cerevisiae and MS2 bacteriophage using acoustic focusing. The acoustic radiation force did not affect the MS2 viruses, and their concentration profile remained unchanged. With optimized design of our microfluidic flow system we were able to achieve yields of > 90% for the MS2 with > 80% of the S. cerevisiae being removed in this continuous-flow sample preparation device.

  18. A random spatial sampling method in a rural developing nation (United States)

    Michelle C. Kondo; Kent D.W. Bream; Frances K. Barg; Charles C. Branas


    Nonrandom sampling of populations in developing nations has limitations and can inaccurately estimate health phenomena, especially among hard-to-reach populations such as rural residents. However, random sampling of rural populations in developing nations can be challenged by incomplete enumeration of the base population. We describe a stratified random sampling method...

  19. A new method for liposome preparation using a membrane contactor. (United States)

    Jaafar-Maalej, Chiraz; Charcosset, Catherine; Fessi, Hatem


    In this article, we present a novel, scalable liposomal preparation technique suitable for the entrapment of pharmaceutical agents into liposomes. This new method is based on the ethanol-injection technique and uses a membrane contactor module, specifically designed for colloidal system preparation. In order to investigate the process, the influence of key parameters on liposome characteristics was studied. It has been established that vesicle-size distribution decreased with a decrease of the organic-phase pressure, an increase of the aqueous-phase flow rate, and a decrease of the phospholipid concentration. Additionally, special attention was paid on reproducibility and long-term stability of lipid vesicles, confirming the robustness of the membrane contactor-based technique. On the other hand, drug-loaded liposomes were prepared and filled with two hydrophobic drug models. High entrapment-efficiency values were successfully achieved for indomethacin (63%) and beclomethasone dipropionate (98%). Transmission electron microscopy images revealed nanometric quasispherical-shaped multilamellar vesicles (size ranging from 50 to 160 nm).

  20. Optimized pre-thinning procedures of ion-beam thinning for TEM sample preparation by magnetorheological polishing. (United States)

    Luo, Hu; Yin, Shaohui; Zhang, Guanhua; Liu, Chunhui; Tang, Qingchun; Guo, Meijian


    Ion-beam-thinning is a well-established sample preparation technique for transmission electron microscopy (TEM), but tedious procedures and labor consuming pre-thinning could seriously reduce its efficiency. In this work, we present a simple pre-thinning technique by using magnetorheological (MR) polishing to replace manual lapping and dimpling, and demonstrate the successful preparation of electron-transparent single crystal silicon samples after MR polishing and single-sided ion milling. Dimples pre-thinned to less than 30 microns and with little mechanical surface damage were repeatedly produced under optimized MR polishing conditions. Samples pre-thinned by both MR polishing and traditional technique were ion-beam thinned from the rear side until perforation, and then observed by optical microscopy and TEM. The results show that the specimen pre-thinned by MR technique was free from dimpling related defects, which were still residual in sample pre-thinned by conventional technique. Nice high-resolution TEM images could be acquired after MR polishing and one side ion-thinning. MR polishing promises to be an adaptable and efficient method for pre-thinning in preparation of TEM specimens, especially for brittle ceramics. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Method for preparing a sodium/sulfur cell (United States)

    Weiner, Steven A.


    A method for preparing a sodium/sulfur cell comprising (A) inserting a solid sodium slug, adapted to be connected to an external circuit, into the anodic reaction zone of a cell subassembly maintained within an inert atmosphere, said cell subassembly comprising a cell container and a tubular cation-permeable barrier disposed within said container such that a first reaction zone is located within cation-permeable barrier and a second reaction zone is located between the outer surface of said cation-permeable barrier and the inner surface of said container, one of said reaction zones being said anodic reaction zone and the other of said reaction zone being a cathodic reaction zone containing a precast composite cathodic reactant comprising a sulfur impregnated porous conductive material connected to said cation permeable barrier and adapted to be connected to said external circuit; and (B) providing closure means for said subassembly and sealing the same to said subassembly at a temperature less than about C. The method of the invention overcomes deficiencies of the prior art methods by allowing preparation of a sodium/sulfur cell without the use of molten reactants and the fill spouts which are required when the cell is filled with molten reactants.

  2. Experience sampling with elderly persons: an exploration of the method. (United States)

    Hnatiuk, S H


    The daily lives of a sample of elderly widows (greater than 69 years of age) were studied using the method of experience sampling developed by Csikszentmihalyi and his colleagues. The purpose of the study was to investigate the response of elderly people to experience sampling as a means of collecting information about their activities, thoughts, and moods during the course of one week. The method proved acceptable to the majority of participants and yielded reliable, valid data about their home lives, particularly from among the younger, more physically able women. Experience sampling was, within certain limits, a useful method of obtaining information from elderly people.


    Directory of Open Access Journals (Sweden)



    Full Text Available Marketing and statistical literature available to practitioners provides a wide range of sampling methods that can be implemented in the context of marketing research. Ranking sampling method is based on taking apart the general population into several strata, namely into several subdivisions which are relatively homogenous regarding a certain characteristic. In fact, the sample will be composed by selecting, from each stratum, a certain number of components (which can be proportional or non-proportional to the size of the stratum until the pre-established volume of the sample is reached. Using ranking sampling within marketing research requires the determination of some relevant statistical indicators - average, dispersion, sampling error etc. To that end, the paper contains a case study which illustrates the actual approach used in order to apply the ranking sample method within a marketing research made by a company which provides Internet connection services, on a particular category of customers – small and medium enterprises.


    It has been over 10 years since the low-flow ground water purging and sampling method was initially reported in the literature. The method grew from the recognition that well purging was necessary to collect representative samples, bailers could not achieve well purging, and high...

  5. Neonatal blood gas sampling methods | Goenka | South African ...

    African Journals Online (AJOL)

    There is little published guidance that systematically evaluates the different methods of neonatal blood gas sampling, where each method has its individual benefits and risks. This review critically surveys the available evidence to generate a comparison between arterial and capillary blood gas sampling, focusing on their ...

  6. An efficient method for sampling the essential subspace of proteins

    NARCIS (Netherlands)

    Amadei, A; Linssen, A.B M; de Groot, B.L.; van Aalten, D.M.F.; Berendsen, H.J.C.

    A method is presented for a more efficient sampling of the configurational space of proteins as compared to conventional sampling techniques such as molecular dynamics. The method is based on the large conformational changes in proteins revealed by the ''essential dynamics'' analysis. A form of

  7. Alterations in cytoskeletal organization and tyrosine phosphorylation in platelet concentrates prepared by the buffy coat method. (United States)

    Estebanell, E; Díaz-Ricart, M; Escolar, G; Lozano, M; Mazzara, R; Ordinas, A


    Numerous morphologic and biochemical changes occurring during platelet storage may result in the impairment of platelet function. The effect of preparation and storage conditions on platelet function was analyzed through evaluation of cytoskeletal organization and signaling mechanisms involved in the activation of platelets by thrombin. Samples of platelets prepared by the buffy coat method were obtained before and after the platelet concentrates were prepared during storage for 1, 3, and 5 days. Thrombin-induced aggregation was monitored, and changes in the organization of proteins in the cytoskeleton were analyzed by gel electrophoresis. For the analysis of tyrosine phosphorylation, proteins were transferred to nitrocellulose membranes and probed with a specific antibody. The aggregation and the cytoskeletal organization induced by thrombin activation were markedly impaired immediately after preparation of platelet concentrates, although they normalized after the first 24 hours of storage and decreased progressively after 3 days of storage. Results in tyrosine phosphorylation paralleled those obtained with cytoskeletal organization, except for samples obtained immediately after processing to obtain platelet concentrates. These data indirectly suggest that the stress induced by the preparation method has an activating effect on platelet function that may imply a delayed platelet response to further stimuli. This effect may result in a deficient redistribution of signaling molecules within platelets.

  8. A method and fortran program for quantitative sampling in paleontology (United States)

    Tipper, J.C.


    The Unit Sampling Method is a binomial sampling method applicable to the study of fauna preserved in rocks too well cemented to be disaggregated. Preliminary estimates of the probability of detecting each group in a single sampling unit can be converted to estimates of the group's volumetric abundance by means of correction curves obtained by a computer simulation technique. This paper describes the technique and gives the FORTRAN program. ?? 1976.

  9. Minerals and antinutrients profile of rabadi after different traditional preparation methods. (United States)

    Gupta, Vineeta; Nagar, Ranjana


    Rabadi is a cereal and buttermilk based traditional fermented recipe of western region of India. There are many traditional preparation methods, which may alter biochemical composition of rabadi, therefore, in the present study, role of traditional processings (cooking, fermentation, dehulling, utensil, preparation methods and cereals) on minerals and antinutrients of pearl millet, wheat flour and refined wheat flour rabadi was investigated on fresh weight basis. Results showed that the process of cooking and fermentation enhanced minerals (Ca, Fe and P) in all types of rabadi samples at different levels of significance, while antinutrients (phytic acid, total phenols and oxalates) reflected a declining trend. Intercomparison of different types of rabadies exhibited that fermented- cooked -fermented samples were better than cooked -fermented rabadies. Dehulling caused a loss of minerals, but antinutrients were also degraded after dehulling; therefore dehulled sample showed very good nutritional profile after fermentation. Earthen pot rabadi samples presented better biochemical composition than rabadies prepared in steel pot. Intercomparison of different cereals based rabadies reflected superior position of fermented -cooked- fermented pearl millet flour rabadi than cooked- fermented pearl millet flour rabadi, wheat and refined wheat flour rabadi samples.

  10. Application of pyrolitic method of synthesis for preparation of calcium bismuthate based photocatalyst (United States)

    Shtarev, Dmitry S.; Makarevich, Konstantin S.; Shtareva, Anna V.; Blokh, Artem I.; Syuy, Alexander V.


    The study describes use of pyrolytic synthesis method for preparation of visible light calcium bismuthate based photocatalyst. The advantage of the proposed method is that it allows obtaining single-phase, homogeneous samples with various types of crystal lattice and controlled defects. It is shown that it is possible to obtain crystalline calcium bismuthate with the same stoichiometry in the entire region of existence of the solid solution. Synthesized samples of calcium bismuthate have been characterized using the scanning electron microscopy and X-ray analysis. It was found that the scope and conditions of synthesis affect the degree of imperfection of both anionic and cationic sublattice of calcium bismuthate.

  11. Structural properties of cobalt substituted barium hexaferrite nanoparticles prepared by a thermal treatment method




    A series of M-type hexagonal ferrites with BaCo xFe 12-xO 19 (x = 2.0, 3.0) composition were synthesized using a simple heat treatment method. The aqueous solution, containing metal nitrates and polyvinyle pyrrolidone (PVP) as a capping agent was used to prepare M-type barium hexferrite nanoparticles. The prepared hexaferrite particles were calcined at microwave frequency (2.45 GHz, power 900W, 5 min) as well as at 650 °C temperature. The structural properties of the samples were investigated...

  12. Whole-Slide Imaging of Pap Cellblock Preparations Is a Potentially Valid Screening Method. (United States)

    Tawfik, Ossama; Davis, Marilyn; Dillon, Susan; Tawfik, Laila; Diaz, Francisco J; Amin, Khalid; Fan, Fang


    To date, the impact of digital imaging on routine cytology remains far from perfect. Cellblock (CB) preparations from Pap samples have been shown to be diagnostically valuable. We evaluated the validity of utilizing whole-slide imaging (WSI) prepared from Pap CBs as a screening tool. A total of 1,110 CB slides prepared from residual Pap samples were analyzed - 563 normal, 282 atypical squamous cells of undetermined significance (ASCUS), 12 atypical squamous cells-cannot exclude high-grade squamous intraepithelial lesion, 188 low-grade squamous intraepithelial lesions (LSIL), 36 high-grade squamous intraepithelial lesions (HSIL), 25 atypical glandular cells of undetermined significance, 1 adenocarcinoma in situ, 2 invasive adenocarcinomas, and 1 squamous cell carcinoma. Virtual slides were obtained using the Aperio system. Test performance characteristics of liquid-based samples and WSI from CB samples were compared. Average sensitivity and specificity of the five WSI reviewers was 58.3 and 85.1% for ASCUS, respectively, 54.1 and 93.9% for LSIL, and 51.8 and 98.8% for HSIL. Overall WSI sensitivity and specificity for detecting lesions was 82.1 and 86.2%, respectively. Agreement (kappa values) between WSI reviewers was 0.56 for ASCUS, 0.69 for LSIL, 0.67 for HSIL, and 0.74 for negative samples. WSI of CB preparations is a feasible method to achieve high-quality specimen preparations. It is as sensitive as liquid-based methods and appears to be highly specific for the detection of LSIL and HSIL. © 2015 S. Karger AG, Basel.

  13. Method and apparatus for imaging a sample on a device (United States)

    Trulson, Mark; Stern, David; Fiekowsky, Peter; Rava, Richard; Walton, Ian; Fodor, Stephen P. A.


    A method and apparatus for imaging a sample are provided. An electromagnetic radiation source generates excitation radiation which is sized by excitation optics to a line. The line is directed at a sample resting on a support and excites a plurality of regions on the sample. Collection optics collect response radiation reflected from the sample I and image the reflected radiation. A detector senses the reflected radiation and is positioned to permit discrimination between radiation reflected from a certain focal plane in the sample and certain other planes within the sample.

  14. On-target sample preparation of 4-sulfophenyl isothiocyanate-derivatized peptides using AnchorChip Targets

    DEFF Research Database (Denmark)

    Zhang, Xumin; Rogowska-Wrzesinska, Adelina; Roepstorff, Peter


    De novo sequencing of tryptic peptides by post source decay (PSD) or collision induced dissociation (CID) analysis using MALDI TOF-TOF instruments is due to the easy interpretation facilitated by the introduction of N-terminal sulfonated derivatives. Recently, a stable and cheap reagent, 4......-sulfophenyl isothiocyanate (SPITC), has been successfully used for N-terminal derivatization. Previously described methods have always used desalting and concentration by reverse-phase chromatography prior to mass spectrometric analysis. Here we present an on-target sample preparation method based on Anchor...

  15. Cytology Preparations of Formalin Fixative Aid Detection of Giardia in Duodenal Biopsy Samples. (United States)

    Panarelli, Nicole C; Gobara, Nariman; Hoda, Rana S; Chaump, Michael; Jessurun, Jose; Yantiss, Rhonda K


    Giardiasis is the most common intestinal parasitic infection in the United States. The organism elicits no, or minimal, inflammatory changes in duodenal biopsy samples, so it can be easily overlooked. We performed this study to determine whether Giardia could be isolated from the formalin fixative of biopsy samples, and to evaluate the value of fluid analysis in the assessment for potential infection. We prospectively evaluated duodenal biopsy samples from 92 patients with a clinical suspicion of giardiasis or symptoms compatible with that diagnosis (ie, diarrhea, bloating, or abdominal pain) Biopsy samples were routinely processed and stained with hematoxylin and eosin. Histologic diagnoses included giardiasis (5 cases, 4%), normal findings (64 cases, 70%), peptic injury/active duodenitis (12 cases, 13%), and intraepithelial lymphocytosis with villous blunting (10 cases, 12%). Fifteen cases (13%) showed detached degenerated epithelial cells or mucus droplets in the intervillous space that resembled Giardia. Cytology slides were prepared from formalin in the biopsy container using the standard Cytospin protocol and reviewed by a cytopathologist blinded to the biopsy findings. Cytologic evaluation revealed Giardia spp. in all 5 biopsy-proven cases, and identified an additional case that was not detected by biopsy analysis. Organisms were significantly more numerous (mean: 400 trophozoites; range, 120 to 810) and showed better morphologic features in cytology preparations compared with tissue sections (mean: 129 trophozoites; range, 37 to 253 organisms; P=0.05). Our findings suggest that cytology preparations from formalin fixative can resolve diagnostically challenging cases and even enhance Giardia detection in some cases.

  16. Evaluating Composite Sampling Methods of Bacillus spores at Low Concentrations

    Energy Technology Data Exchange (ETDEWEB)

    Hess, Becky M.; Amidan, Brett G.; Anderson, Kevin K.; Hutchison, Janine R.


    Restoring facility operations after the 2001 Amerithrax attacks took over three months to complete, highlighting the need to reduce remediation time. The most time intensive tasks were environmental sampling and sample analyses. Composite sampling allows disparate samples to be combined, with only a single analysis needed, making it a promising method to reduce response times. We developed a statistical experimental design to test three different composite sampling methods: 1) single medium single pass composite: a single cellulose sponge samples multiple coupons; 2) single medium multi-pass composite: a single cellulose sponge is used to sample multiple coupons; and 3) multi-medium post-sample composite: a single cellulose sponge samples a single surface, and then multiple sponges are combined during sample extraction. Five spore concentrations of Bacillus atrophaeus Nakamura spores were tested; concentrations ranged from 5 to 100 CFU/coupon (0.00775 to 0.155CFU/cm2, respectively). Study variables included four clean surface materials (stainless steel, vinyl tile, ceramic tile, and painted wallboard) and three grime coated/dirty materials (stainless steel, vinyl tile, and ceramic tile). Analysis of variance for the clean study showed two significant factors: composite method (p-value < 0.0001) and coupon material (p-value = 0.0008). Recovery efficiency (RE) was higher overall using the post-sample composite (PSC) method compared to single medium composite from both clean and grime coated materials. RE with the PSC method for concentrations tested (10 to 100 CFU/coupon) was similar for ceramic tile, painted wall board, and stainless steel for clean materials. RE was lowest for vinyl tile with both composite methods. Statistical tests for the dirty study showed RE was significantly higher for vinyl and stainless steel materials, but significantly lower for ceramic tile. These results suggest post-sample compositing can be used to reduce sample analysis time when

  17. Calcium Deficient Hydroxyapatite for Medical Application Prepared by Hydrothermal Method (United States)

    Ioku, Koji; Kamitakahara, Masanobu; Ikeda, Tohru


    Hydrothermal processing plays a key role in the synthesis of biomaterials with excellent biocompatibility in the physiological environment. Especially, calcium phosphates are paid to much attention for the regenerative medicine. Two kinds of porous materials of hydroxyapatite with 70% porosity were prepared. One of them is a newly developed calcium-deficient hydroxyapatite composed of rod-shaped particles of about 20 μm in length synthesized hydrothermally (HHA) and the other one is the stoichiometric hydroxyapatite (SHA) prepared by the conventional sintering method. These materials were used for animal implantation tests to compare these biological responses. In the rabbit femur, implanted HHA was slowly resorbed and then most of the implanted HHA was resorbed after 72 weeks. The implanted SHA was unresorbed throughout the experimental period. The volume of newly formed bone and the number of osteoclasts in the implanted region were significantly larger in HHA than in SHA after 24 weeks. Results in the present research suggested that the activity of osteoclasts correlated to the bone forming activity of osteoblasts. The method to synthesize biodegradable pure calcium-deficient HA is expected to provide adequate biodegradability and bone replaceability.

  18. Sampling Methods for Web and E-mail Surveys


    Fricker, RD


    London: SAGE Publications. Reprinted from The SAGE Handbook of Online Research Methods, N. Fielding, R.M. Lee and G. Blank, eds., chapter 11, London: SAGE Publications, 195-216. This chapter is a comprehensive overview of sampling methods for web and e-mail (‘Internetbased’) surveys. It reviews the various types of sampling method – both probability and nonprobability – and examines their applicability to Internet-based surveys. Issues related to Internetbased survey samp...

  19. Evaluation of common methods for sampling invertebrate pollinator assemblages: net sampling out-perform pan traps.

    Directory of Open Access Journals (Sweden)

    Tony J Popic

    Full Text Available Methods for sampling ecological assemblages strive to be efficient, repeatable, and representative. Unknowingly, common methods may be limited in terms of revealing species function and so of less value for comparative studies. The global decline in pollination services has stimulated surveys of flower-visiting invertebrates, using pan traps and net sampling. We explore the relative merits of these two methods in terms of species discovery, quantifying abundance, function, and composition, and responses of species to changing floral resources. Using a spatially-nested design we sampled across a 5000 km(2 area of arid grasslands, including 432 hours of net sampling and 1296 pan trap-days, between June 2010 and July 2011. Net sampling yielded 22% more species and 30% higher abundance than pan traps, and better reflected the spatio-temporal variation of floral resources. Species composition differed significantly between methods; from 436 total species, 25% were sampled by both methods, 50% only by nets, and the remaining 25% only by pans. Apart from being less comprehensive, if pan traps do not sample flower-visitors, the link to pollination is questionable. By contrast, net sampling functionally linked species to pollination through behavioural observations of flower-visitation interaction frequency. Netted specimens are also necessary for evidence of pollen transport. Benefits of net-based sampling outweighed minor differences in overall sampling effort. As pan traps and net sampling methods are not equivalent for sampling invertebrate-flower interactions, we recommend net sampling of invertebrate pollinator assemblages, especially if datasets are intended to document declines in pollination and guide measures to retain this important ecosystem service.

  20. A novel method for preparing and characterizing alcoholic EPD suspensions. (United States)

    De Riccardis, M F; Carbone, D; Rizzo, A


    Ceramic suspensions composed of alumina and mixtures of alumina and zirconia powders in ethyl alcohol were prepared. A solution of citric acid and triethylamine was used as dispersant. The citric acid, which usually is used as dispersant in water alumina suspensions, gave excellent results in ethyl alcohol also if it was used in conjunction with triethylamine. A novel method consisting of combined measurements of grain size, zeta potential, and transmittance was optimized to study the dispersion and stability properties of the ceramic suspensions; by using this method the optimal dispersant amount was determined. The suspensions based on alumina and alumina-zirconia powders were used to coat stainless steel plates by electrophoretic deposition (EPD); the optimal composition of suspensions and the used EPD parameters made it possible to obtain coatings with uniform thickness and composition.

  1. A combined method for correlative 3D imaging of biological samples from macro to nano scale (United States)

    Kellner, Manuela; Heidrich, Marko; Lorbeer, Raoul-Amadeus; Antonopoulos, Georgios C.; Knudsen, Lars; Wrede, Christoph; Izykowski, Nicole; Grothausmann, Roman; Jonigk, Danny; Ochs, Matthias; Ripken, Tammo; Kühnel, Mark P.; Meyer, Heiko


    Correlative analysis requires examination of a specimen from macro to nano scale as well as applicability of analytical methods ranging from morphological to molecular. Accomplishing this with one and the same sample is laborious at best, due to deformation and biodegradation during measurements or intermediary preparation steps. Furthermore, data alignment using differing imaging techniques turns out to be a complex task, which considerably complicates the interconnection of results. We present correlative imaging of the accessory rat lung lobe by combining a modified Scanning Laser Optical Tomography (SLOT) setup with a specially developed sample preparation method (CRISTAL). CRISTAL is a resin-based embedding method that optically clears the specimen while allowing sectioning and preventing degradation. We applied and correlated SLOT with Multi Photon Microscopy, histological and immunofluorescence analysis as well as Transmission Electron Microscopy, all in the same sample. Thus, combining CRISTAL with SLOT enables the correlative utilization of a vast variety of imaging techniques.

  2. An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology (United States)

    Thompson, Rebecca F.; Walker, Matt; Siebert, C. Alistair; Muench, Stephen P.; Ranson, Neil A.


    Transmission electron microscopy (EM) is a versatile technique that can be used to image biological specimens ranging from intact eukaryotic cells to individual proteins >150 kDa. There are several strategies for preparing samples for imaging by EM, including negative staining and cryogenic freezing. In the last few years, cryo-EM has undergone a ‘resolution revolution’, owing to both advances in imaging hardware, image processing software, and improvements in sample preparation, leading to growing number of researchers using cryo-EM as a research tool. However, cryo-EM is still a rapidly growing field, with unique challenges. Here, we summarise considerations for imaging of a range of specimens from macromolecular complexes to cells using EM. PMID:26931652

  3. Determination of aglycones of ginsenosides in ginseng preparations sold in Sweden and in urine samples from Swedish athletes consuming ginseng. (United States)

    Cui, J F; Garle, M; Björkhem, I; Eneroth, P


    Recently developed gas chromatographic and gas chromatographic-mass spectrometric methods were used to characterize 17 different commercial ginseng preparations sold in Sweden. The contents of total ginsenosides per capsule or per tablet varied from 2.1 to 13.3 mg. Unlike the other preparations, a red ginseng and three liquid ginseng preparations (after releasing the sugar moieties from ginsenosides) were shown also to contain significant amounts of 20-epimers of 20(S)-protopanaxadiol and 20(S)-protopanaxatriol as well as their corresponding 24,25-hydrated compounds. In addition to the genuine and artificial sapogenins mentioned above, two epimeric pairs of prosapogenines (ginsenoside Rg3 and 20(S)-Rg3, ginsenoside Rh1 and 20(R)-Rh1) were also found in the liquid formulations. These results suggest that hydrolysis, epimerization and hydration in the side-chain of the aglycone moiety of ginsenosides may occur in the liquid formulations under weak acidic conditions (pH 3.0-3.5 with 9-10% of alcohol at room temperature). The new method was also used to determine the aglycones of ginsenosides in urine samples from Swedish athletes stating that they had consumed ginseng preparations within 10 days before urine collection. Out of the 65 samples analysed, 60 were found to contain 20(S)-protopanaxatriol. The concentrations of 20(S)-protopanaxatriol ginsenosides varied from 2 to 35 ng ml-1 urine. This is the first demonstration of uptake of ginsenosides in humans after oral administration of ginseng preparations.

  4. An investigation into the sample preparation procedure and analysis of cyanoacrylate adhesives using capillary electrophoresis


    Whitaker, Gillian; Kincaid, Brendan J.; Van Hoof, Nicole; Regan, Fiona; Smyth, Malcolm R.; Leonard, Raymond G.


    In this study, the trace acid profile of cyanoacrylate adhesives was studied using capillary electrophoresis. Liquid–liquid extraction was employed as the sample preparation step before separation by capillary electrophoresis. The solubility of the adhesives was investigated using various organic solvents, e.g. hexane and dichloromethane, and chloroform was determined to be the optimum solvent as it enabled the full dissolution of the adhesive. A comprehensive stability study was performed ov...

  5. Chitosan-Based Matrices Prepared by Gamma Irradiation for Tissue Regeneration: Structural Properties vs. Preparation Method. (United States)

    Casimiro, Maria Helena; Lancastre, Joana J H; Rodrigues, Alexandra P; Gomes, Susana R; Rodrigues, Gabriela; Ferreira, Luís M


    In the last decade, new generations of biopolymer-based materials have attracted attention, aiming its application as scaffolds for tissue engineering. These engineered three-dimensional scaffolds are designed to improve or replace damaged, missing, or otherwise compromised tissues or organs. Despite the number of promising methods that can be used to generate 3D cell-instructive matrices, the innovative nature of the present work relies on the application of ionizing radiation technology to form and modify surfaces and matrices with advantage over more conventional technologies (room temperature reaction, absence of harmful initiators or solvents, high penetration through the bulk materials, etc.), and the possibility of preparation and sterilization in one single step. The current chapter summarizes the work done by the authors in the gamma radiation processing of biocompatible and biodegradable chitosan-based matrices for skin regeneration. Particular attention is given to the correlation between the different preparation conditions and the final polymeric matrices' properties. We therefore expect to demonstrate that instructive matrices produced and improved by radiation technology bring to the field of skin regenerative medicine a supplemental advantage over more conservative techniques.

  6. Preparation, microstructure and properties of yttrium aluminum garnet fibers prepared by sol-gel method

    Energy Technology Data Exchange (ETDEWEB)

    Li Chengshun [Key Laboratory for Liquid Structure and Heredity of Materials of Ministry of Education, Shandong University, Jinan 250061 (China); Zhang Yujun [Key Laboratory for Liquid Structure and Heredity of Materials of Ministry of Education, Shandong University, Jinan 250061 (China)], E-mail:; Gong Hongyu; Zhang Jingde; Nie Lifang [Key Laboratory for Liquid Structure and Heredity of Materials of Ministry of Education, Shandong University, Jinan 250061 (China)


    Yttrium aluminum garnet (YAG) fiber was prepared by sol-gel method using water as the solvent. The spinnable YAG sol was synthesized using Al powder, Y(CH{sub 3}COOH){sub 3}.4H{sub 2}O and HCl as precursors, polyethylene oxide as viscosity adjusting agent. Gel fibers with diameter of 5-10 {mu}m were prepared from the YAG sol by using centrifugal spinning technique. YAG crystalline fibers were obtained by drying gel fibers and heat-treating at selected temperature. TG/DTA analysis showed an exotherm at 906 deg. C attributed to formation of YAG phase and weight loss of 45% at 1000 deg. C. XRD and FT-IR analysis showed that phase-pure YAG can be formed at 900 deg. C, and no other intermediate was observed. The grain size of YAG fibers increased from 25 to 220 nm and tensile strength decreased rapidly from 970 to 380 MPa when the sintering temperature increased from 900 to 1550 deg. C.

  7. An Automated Sample Preparation System for Large-Scale DNA Sequencing (United States)

    Marziali, Andre; Willis, Thomas D.; Federspiel, Nancy A.; Davis, Ronald W.


    Recent advances in DNA sequencing technologies, both in the form of high lane-density gels and automated capillary systems, will lead to an increased requirement for sample preparation systems that operate at low cost and high throughput. As part of the development of a fully automated sequencing system, we have developed an automated subsystem capable of producing 10,000 sequence-ready ssDNA templates per day from libraries of M13 plaques at a cost of $0.29 per sample. This Front End has been in high throughput operation since June, 1997 and has produced > 400,000 high-quality DNA templates. PMID:10330125

  8. Simulation of topography evolution and damage formation during TEM sample preparation using focused ion beams

    Energy Technology Data Exchange (ETDEWEB)

    Boxleitner, W. E-mail:; Hobler, G.; Klueppel, V.; Cerva, H


    Our recently developed simulation code FIBSIM is applied to topics related to transmission electron microscopy (TEM) sample preparation using focused ion beams (FIB). FIBSIM combines dynamic Monte Carlo simulation of collision cascades with two-dimensional, cell-based topography simulation. The influence of the scanning mode and of the beam current profile on the evolution of the surface contour is investigated. Furthermore, amorphous zones in silicon samples and damaged regions are predicted for different beam energies of 10, 30 and 50 keV. The thickness of the predicted amorphous regions is in good agreement with experimental TEM data.

  9. Technical Note: An improved guideline for rapid and precise sample preparation of tree-ring stable isotope analysis (United States)

    Schollaen, K.; Baschek, H.; Heinrich, I.; Helle, G.


    The procedure of wood sample preparation, including tree-ring dissection, cellulose extraction, homogenization and finally weighing and packing for stable isotope analysis is labour intensive and time consuming. We present an elaborated methodical guideline from pre-analyses considerations, wood sample preparation through semi-automated chemical extraction of cellulose directly from tree-ring cross-sections to tree-ring dissection for high-precision isotope ratio mass spectrometry. This guideline reduces time and maximizes the tree-ring stable isotope data throughput significantly. The method was applied to ten different tree species (coniferous and angiosperm wood) with different wood growth rates and differently shaped tree-ring boundaries. The tree-ring structures of the cellulose cross-sections largely remained well identifiable. FTIR (Fourier transform infrared) spectrometry and the comparison of stable isotope values with classical method confirm chemical purity of the resultant cellulose. Sample homogenization is no longer necessary. Cellulose extraction is now faster, cheaper and more user friendly allowing (i) the simultaneous treatment of wood cross-sections of a total length of 180 cm (equivalent to 6 increment cores of 30 cm length) and thickness of 0.5 to 2 mm, and (ii) precise tree-ring separation at annual to high-resolution scale utilizing manual devices or UV-laser microdissection microscopes.

  10. Method for using polarization gating to measure a scattering sample (United States)

    Baba, Justin S.


    Described herein are systems, devices, and methods facilitating optical characterization of scattering samples. A polarized optical beam can be directed to pass through a sample to be tested. The optical beam exiting the sample can then be analyzed to determine its degree of polarization, from which other properties of the sample can be determined. In some cases, an apparatus can include a source of an optical beam, an input polarizer, a sample, an output polarizer, and a photodetector. In some cases, a signal from a photodetector can be processed through attenuation, variable offset, and variable gain.

  11. Reproducibility of NMR Analysis of Urine Samples: Impact of Sample Preparation, Storage Conditions, and Animal Health Status

    Directory of Open Access Journals (Sweden)

    Christina Schreier


    Full Text Available Introduction. Spectroscopic analysis of urine samples from laboratory animals can be used to predict the efficacy and side effects of drugs. This employs methods combining 1H NMR spectroscopy with quantification of biomarkers or with multivariate data analysis. The most critical steps in data evaluation are analytical reproducibility of NMR data (collection, storage, and processing and the health status of the animals, which may influence urine pH and osmolarity. Methods. We treated rats with a solvent, a diuretic, or a nephrotoxicant and collected urine samples. Samples were titrated to pH 3 to 9, or salt concentrations increased up to 20-fold. The effects of storage conditions and freeze-thaw cycles were monitored. Selected metabolites and multivariate data analysis were evaluated after 1H NMR spectroscopy. Results. We showed that variation of pH from 3 to 9 and increases in osmolarity up to 6-fold had no effect on the quantification of the metabolites or on multivariate data analysis. Storage led to changes after 14 days at 4°C or after 12 months at −20°C, independent of sample composition. Multiple freeze-thaw cycles did not affect data analysis. Conclusion. Reproducibility of NMR measurements is not dependent on sample composition under physiological or pathological conditions.

  12. Preparation of silica by sol-gel method using formamide

    Directory of Open Access Journals (Sweden)

    R.F.S. Lenza


    Full Text Available In this work we obtained microporous and mesoporous silica gels by sol-gel processing. Tetraethylortosilicate (TEOS was used as precursor. Nitric acid and hydrofluoric acid were used as catalysts. In order to study the affect of formamide as drying additive, we used a molar ratio alkoxide/formamide of 1/1. The performance of formamide in obtaining crack-free gels was evaluated through monolithicity measurements. The structural evolution occurring in the interconnected network of the gels during thermal treatment was monitored by Fourier transform infrared spectroscopy (FTIR, shrinkage and density measurements and nitrogen gas sorption. We noted that in the presence of formamide, the Si-O-Si bonds are stronger and belong to a more cross-linked structure. The samples obtained in the presence of formamide have larger pore volume and its pore structure is in the range of mesoporosity. The samples obtained without additive are microporous. Formamide allowed the preparation of crack-free silica gels stabilized at high temperatures.

  13. [Respondent-Driven Sampling: a new sampling method to study visible and hidden populations]. (United States)

    Mantecón, Alejandro; Juan, Montse; Calafat, Amador; Becoña, Elisardo; Román, Encarna


    The paper introduces a variant of chain-referral sampling: respondent-driven sampling (RDS). This sampling method shows that methods based on network analysis can be combined with the statistical validity of standard probability sampling methods. In this sense, RDS appears to be a mathematical improvement of snowball sampling oriented to the study of hidden populations. However, we try to prove its validity with populations that are not within a sampling frame but can nonetheless be contacted without difficulty. The basics of RDS are explained through our research on young people (aged 14 to 25) who go clubbing, consume alcohol and other drugs, and have sex. Fieldwork was carried out between May and July 2007 in three Spanish regions: Baleares, Galicia and Comunidad Valenciana. The presentation of the study shows the utility of this type of sampling when the population is accessible but there is a difficulty deriving from the lack of a sampling frame. However, the sample obtained is not a random representative one in statistical terms of the target population. It must be acknowledged that the final sample is representative of a 'pseudo-population' that approximates to the target population but is not identical to it.

  14. Sampling and analysis methods for geothermal fluids and gases

    Energy Technology Data Exchange (ETDEWEB)

    Watson, J.C.


    The sampling procedures for geothermal fluids and gases include: sampling hot springs, fumaroles, etc.; sampling condensed brine and entrained gases; sampling steam-lines; low pressure separator systems; high pressure separator systems; two-phase sampling; downhole samplers; and miscellaneous methods. The recommended analytical methods compiled here cover physical properties, dissolved solids, and dissolved and entrained gases. The sequences of methods listed for each parameter are: wet chemical, gravimetric, colorimetric, electrode, atomic absorption, flame emission, x-ray fluorescence, inductively coupled plasma-atomic emission spectroscopy, ion exchange chromatography, spark source mass spectrometry, neutron activation analysis, and emission spectrometry. Material on correction of brine component concentrations for steam loss during flashing is presented. (MHR)

  15. [Biological Advisory Subcommittee Sampling Methods : Results, Resolutions, and Correspondences : 2002 (United States)

    US Fish and Wildlife Service, Department of the Interior — This document contains a variety of information concerning Biological Advisory Subcommittee sampling methods at the Rocky Mountain Arsenal Refuge in 2002. Multiple...

  16. A cryopreservation method for Pasteurella multocida from wetland samples (United States)

    Moore, Melody K.; Shadduck, D.J.; Goldberg, D.R.; Samuel, M.D.


    A cryopreservation method and improved isolation techniques for detection of Pasteurella multocida from wetland samples were developed. Wetland water samples were collected in the field, diluted in dimethyl sulfoxide (DMSO, final concentration 10%), and frozen at -180 C in a liquid nitrogen vapor shipper. Frozen samples were transported to the laboratory where they were subsequently thawed and processed in Pasteurella multocida selective broth (PMSB) to isolate P. multocida. This method allowed for consistent isolation of 2 to 18 organisms/ml from water seeded with known concentrations of P. multocida. The method compared favorably with the standard mouse inoculation method and allowed for preservation of the samples until they could be processed in the laboratory.

  17. Methods for collection and analysis of water samples (United States)

    Rainwater, Frank Hays; Thatcher, Leland Lincoln


    This manual contains methods used by the U.S. Geological Survey to collect, preserve, and analyze water samples. Throughout, the emphasis is on obtaining analytical results that accurately describe the chemical composition of the water in situ. Among the topics discussed are selection of sampling sites, frequency of sampling, field equipment, preservatives and fixatives, analytical techniques of water analysis, and instruments. Seventy-seven laboratory and field procedures are given for determining fifty-three water properties.

  18. Field evaluation of personal sampling methods for multiple bioaerosols.

    Directory of Open Access Journals (Sweden)

    Chi-Hsun Wang

    Full Text Available Ambient bioaerosols are ubiquitous in the daily environment and can affect health in various ways. However, few studies have been conducted to comprehensively evaluate personal bioaerosol exposure in occupational and indoor environments because of the complex composition of bioaerosols and the lack of standardized sampling/analysis methods. We conducted a study to determine the most efficient collection/analysis method for the personal exposure assessment of multiple bioaerosols. The sampling efficiencies of three filters and four samplers were compared. According to our results, polycarbonate (PC filters had the highest relative efficiency, particularly for bacteria. Side-by-side sampling was conducted to evaluate the three filter samplers (with PC filters and the NIOSH Personal Bioaerosol Cyclone Sampler. According to the results, the Button Aerosol Sampler and the IOM Inhalable Dust Sampler had the highest relative efficiencies for fungi and bacteria, followed by the NIOSH sampler. Personal sampling was performed in a pig farm to assess occupational bioaerosol exposure and to evaluate the sampling/analysis methods. The Button and IOM samplers yielded a similar performance for personal bioaerosol sampling at the pig farm. However, the Button sampler is more likely to be clogged at high airborne dust concentrations because of its higher flow rate (4 L/min. Therefore, the IOM sampler is a more appropriate choice for performing personal sampling in environments with high dust levels. In summary, the Button and IOM samplers with PC filters are efficient sampling/analysis methods for the personal exposure assessment of multiple bioaerosols.

  19. Adaptive cluster sampling: An efficient method for assessing inconspicuous species (United States)

    Andrea M. Silletti; Joan Walker


    Restorationistis typically evaluate the success of a project by estimating the population sizes of species that have been planted or seeded. Because total census is raely feasible, they must rely on sampling methods for population estimates. However, traditional random sampling designs may be inefficient for species that, for one reason or another, are challenging to...

  20. Methods of Fire Debris Preparation for Detection of Accelerants. (United States)

    Caddy, B; Smith, F P; Macy, J


    Forensic scientists use various techniques to separate accelerants from fire debris samples before instrumental identification of added fuels. Among the choices available, traditional micro-distillation, steam distillation, vacuum distillation, headspace, heated headspace, and several vapor adsorption/desorption methods provide various advantages and disadvantages. This communication reviews the development of these techniques from the 1950s and comparison studies performed. Copyright © 1991 Central Police University.

  1. Hydroxyapatite-phosphonoformic acid hybrid compounds prepared by hydrothermal method (United States)

    Turki, Thouraya; Othmani, Masseoud; Bantignies, Jean-Louis; Bouzouita, Khaled


    Hydroxyapatites were prepared in the presence of different amounts of phosphonoformic acid (PFA) via the hydrothermal method. The obtained powders were characterized through chemical analysis, XRD, IR, 31P MAS-NMR, TEM, and TG-TDA. The XRD showed that the PFA did not affect the apatite composition. Indeed, only a reduction of the crystallite size was noted. After grafting of PFA, the IR spectroscopy revealed the appearance of new bands belonging to HPO42- and carboxylate groups of the apatite and organic moiety, respectively. Moreover, the 31P MAS-NMR spectra exhibited a peak with a low intensity assigned to the terminal phosphonate group of the organic moiety in addition to that of the apatite. Based on these results, a reaction mechanism involving the surface hydroxyl groups (tbnd Casbnd OH) of the apatite and the carboxyl group of the acid was proposed.

  2. Method for preparing dioxyheterocycle-based electrochromic polymers

    Energy Technology Data Exchange (ETDEWEB)

    Reynolds, John R.; Estrada, Leandro; Deininger, James; Arroyave-Mondragon, Frank Antonio


    A method for preparing a conjugated polymer involves a DHAP polymerization of a 3,4-dioxythiophene, 3,4-dioxyfuran, or 3,4-dioxypyrrole and, optionally, at least one second conjugated monomer in the presence of a Pd or Ni comprising catalyst, an aprotic solvent, a carboxylic acid at a temperature in excess of C. At least one of the monomers is substituted with hydrogen reactive functionalities and at least one of the monomers is substituted with a Cl, Br, and/or I. The polymerization can be carried out at temperature of C. or more, and the DHAP polymerization can be carried out without a phosphine ligand or a phase transfer agent. The resulting polymer can display dispersity less than 2 and have a degree of polymerization in excess of 10.


    Directory of Open Access Journals (Sweden)

    Suo Xia Hou


    Full Text Available Nano molybdenum disulfide possesses unique chemical and physical properties. In this paper molybdenum disulfide nanoparticles with spherical and flower-like structure are prepared via a hydrothermal method. Sodium molybdate and thioacetamide are taken as precursors, polyethylene glycol (PEG-20000, hexadecyl trimethyl ammonium chloride (CTAC and anhydrous ethanol are used as additives. The properties of the product are characterized by X-ray diffraction (XRD and scanning electron microscopy (SEM. The results showed that under acidic conditions, molybdenum disulfide nanoparticles with spherical shape are obtained when PEG-20000 and CTAC are added. The nanoparticles are uniform in size with a diameter of about 100 nm. Molybdenum disulfide nanoparticles with a flower-like structure are obtained when anhydrous ethanol is added. Their diameters under sulfuric acid and hydrochloric acid conditions are 190 nm and 70 nm, respectively. Yield analysis reveals that the highest yield (which can be up to 79 % occurs by adding polyethylene glycol in a sulfuric acid environment.

  4. Preparo de amostras e métodos para a determinação do teor de óleo essencial de frutos de limoeiro Samples preparation form and methods of analysis for determination of essential oil content on lemon fruits

    Directory of Open Access Journals (Sweden)

    Hélio Grassi Filho


    Full Text Available Objetivou-se avaliar alternativas de preparo da amostra e os métodos quantitativos para a determinação do teor de óleo em frutos de limão. Os tratamentos foram avaliados por meio do delineamento estatístico inteiramente casualizado, em esquema fatorial 2x2x6, com quatro repetições, sendo duas variedades-copa de limoeiro verdadeiro ('Siciliano' e 'Eureka', dois métodos analíticos para a determinação do teor de óleo (Scott e Clevenger e seis formas de preparo dos frutos para análise (análise do fruto inteiro; frutos cortados na longitudinal e análise de ¼; frutos cortados na longitudinal e análise de ¹/2; frutos cortados ao meio e análise da parte superior; frutos cortados ao meio e análise da parte inferior; e análise de ²/8 do fruto. O método denominado de Clevenger foi o mais eficiente na determinação do teor de óleo essencial, e as formas de preparo de amostras ½ inferior, ½ superior, ¼ longitudinal e ½ longitudinal dos frutos proporcionaram valores superiores de óleo essencial.The aim of this research was to evaluate the samples preparation form and methods of analysis to determinate the essential oil content in 'Siciliano' and 'Eureka' lemon fruits. The better sample form was determinated using the fresh and mature fruit, as following: analysis of full fruits, fruits cut in longitudinal way and analysis of ¹/4 of this one, fruits cut in longitudinal way and analysis of the half, fruits cut in the middle and superior part analysis, fruits cut in the middle and inferior part analysis and ²/8 of fruits analysis. The treatments were evaluated by a completely randomized experimental design, in a factorial scheme 2x2x6, with four replications. It was concluded that the Clevenger method was more efficient on the determination of the oil content, and the fruits cut in the ½ inferior, ½ superior, ¼ longitudinal and ½ longitudinal way provided superior values of essential oil content.

  5. Evaluation of grinding methods for pellets preparation aiming at the analysis of plant materials by laser induced breakdown spectrometry


    Gomes, Marcos da Silva; Santos Junior, Dario [UNIFESP; Nunes, Lidiane Cristina; Arantes de Carvalho, Gabriel Gustinelli; Leme, Flavio de Oliveira; Krug, Francisco Jose


    It has been demonstrated that laser induced breakdown spectrometry (LIBS) can be used as an alternative method for the determination of macro (P, K. Ca, Mg) and micronutrients (B, Fe, Cu, Mn, Zn) in pellets of plant materials. However, information is required regarding the sample preparation for plant analysis by LIBS. in this work, methods involving cryogenic grinding and planetary ball milling were evaluated for leaves comminution before pellets preparation. the particle sizes were associat...

  6. DOE methods for evaluating environmental and waste management samples

    Energy Technology Data Exchange (ETDEWEB)

    Goheen, S.C.; McCulloch, M.; Thomas, B.L.; Riley, R.G.; Sklarew, D.S.; Mong, G.M.; Fadeff, S.K. [eds.


    DOE Methods for Evaluating Environmental and Waste Management Samples (DOE Methods) is a resource intended to support sampling and analytical activities for the evaluation of environmental and waste management samples from U.S. Department of Energy (DOE) sites. DOE Methods is the result of extensive cooperation from all DOE analytical laboratories. All of these laboratories have contributed key information and provided technical reviews as well as significant moral support leading to the success of this document. DOE Methods is designed to encompass methods for collecting representative samples and for determining the radioisotope activity and organic and inorganic composition of a sample. These determinations will aid in defining the type and breadth of contamination and thus determine the extent of environmental restoration or waste management actions needed, as defined by the DOE, the U.S. Environmental Protection Agency, or others. The development of DOE Methods is supported by the Analytical Services Division of DOE. Unique methods or methods consolidated from similar procedures in the DOE Procedures Database are selected for potential inclusion in this document. Initial selection is based largely on DOE needs and procedure applicability and completeness. Methods appearing in this document are one of two types, {open_quotes}Draft{close_quotes} or {open_quotes}Verified{close_quotes}. {open_quotes}Draft{close_quotes} methods that have been reviewed internally and show potential for eventual verification are included in this document, but they have not been reviewed externally, and their precision and bias may not be known. {open_quotes}Verified{close_quotes} methods in DOE Methods have been reviewed by volunteers from various DOE sites and private corporations. These methods have delineated measures of precision and accuracy.

  7. Soybean and lactose in meat products and preparations sampled at retail

    Directory of Open Access Journals (Sweden)

    Filomena Piccolo


    Full Text Available Food allergies and intolerances have increased during the last decades and regulatory authorities have taken different measures to prevent and manage consumers’ adverse reactions, including correct labelling of foods. Aim of this work was to search for soybean and lactose in meat products and meat preparations taken from retail in some provinces of Campania Region (Southern Italy and to evaluate the food labels compliance with Regulation (EU n.1169/2011. Soybean and lactose were searched using commercial kits in n. 58 samples of meat products produced in or distributed by 19 establishments, and in n. 55 samples of meat products and n. 8 of meat preparations produced in 21 plants. All samples were selected on the basis of the absence of any information on the labels about the presence of the two searched allergens, with the exception of n. 5 samples tested for lactose. Traces of soybean were detected in 50 out of the 58 examined samples, at concentrations up to 0.93 mg kg–1. Only two samples contained levels above the detection limit of 0.31 mg kg–1. Lactose levels ranging from 0.11 to 2.95 g/100 g, i.e. above the detection limit, were found in all the tested samples (n. 63. The results of the present research underline the need for careful controls and planning by operators as part of the self-control plans, and deserve attention from the competent authorities considering not only the consumers’ health but also the great attention media pay to regulations providing consumers with information on food.

  8. Method and sample spinning apparatus for measuring the NMR spectrum of an orientationally disordered sample (United States)

    Pines, Alexander; Samoson, Ago


    An improved NMR apparatus and method are described which substantially improve the resolution of NMR measurements made on powdered or amorphous or otherwise orientationally disordered samples. The apparatus spins the sample about an axis. The angle of the axis is mechanically varied such that the time average of two or more Legendre polynomials are zero.

  9. Methods for sample size determination in cluster randomized trials. (United States)

    Rutterford, Clare; Copas, Andrew; Eldridge, Sandra


    The use of cluster randomized trials (CRTs) is increasing, along with the variety in their design and analysis. The simplest approach for their sample size calculation is to calculate the sample size assuming individual randomization and inflate this by a design effect to account for randomization by cluster. The assumptions of a simple design effect may not always be met; alternative or more complicated approaches are required. We summarise a wide range of sample size methods available for cluster randomized trials. For those familiar with sample size calculations for individually randomized trials but with less experience in the clustered case, this manuscript provides formulae for a wide range of scenarios with associated explanation and recommendations. For those with more experience, comprehensive summaries are provided that allow quick identification of methods for a given design, outcome and analysis method. We present first those methods applicable to the simplest two-arm, parallel group, completely randomized design followed by methods that incorporate deviations from this design such as: variability in cluster sizes; attrition; non-compliance; or the inclusion of baseline covariates or repeated measures. The paper concludes with methods for alternative designs. There is a large amount of methodology available for sample size calculations in CRTs. This paper gives the most comprehensive description of published methodology for sample size calculation and provides an important resource for those designing these trials. © The Author 2015. Published by Oxford University Press on behalf of the International Epidemiological Association.

  10. Methods of preparation of Swazi traditional fermented foods

    Directory of Open Access Journals (Sweden)

    Protus Simatende


    Conclusion: Umcombotsi, emahewu, buganu, and emasi were the fermented foods commonly prepared at a household level in the Hhohho region, Swaziland. The main ingredient used for preparing umcombotsi and emahewu was maize meal. Unmilled sorghum malt was also added during preparation of umcombotsi. However, typically no malt was added during the preparation of emahewu. Buganu and emasi also play an important role in the diet and socioeconomic activities of the population in Swaziland.

  11. Preventing and Removing Contamination in a Natural Radiocarbon Sample Preparation Laboratory

    Energy Technology Data Exchange (ETDEWEB)

    Zermeno, P; Kurdyla, D K; Buchholz, B A; Heller, S J; Frantz, B R; Brown, T A; Kashgarian, M


    The introduction of elevated {sup 14}C contamination into a natural radiocarbon sample preparation laboratory can occur through many different pathways. The most difficult to control is the introduction of contaminated samples from outside labs. Laboratories can remain {sup 14}C contaminated as a result of earlier tracer based research, even if ''hot'' work has not occurred in the laboratories in decades. Prior to accepting samples from outside collaborators, it is recommended that the collaborators test their labs for {sup 14}C contamination. Any surface in a lab that has high use by multiple people has the potential to be contaminated. The standard procedure for determining whether a collaborator's lab is contaminated consists of swiping lab surfaces with small glass fiber filters wetted with alcohol and measuring them for {sup 14}C content using AMS. Volatile {sup 14}C can be detected by using aerosol monitors consisting of fine soot that is depleted in {sup 14}C. These monitors can be set out in the laboratory in question to check for volatile {sup 14}C contamination. In the event that a hot sample is introduced in the natural radiocarbon sample prep laboratory, all sample submission should be stopped until the lab is declared clean. Samples already being processed should be completed along with {sup 14}C depleted material and measured by AMS. This will help determine if the contaminated samples have affected other samples in the laboratory. After a contamination event, the laboratory and associated equipment requires cleaning or disposal. All surfaces and equipment should be wiped down with acetone or ethanol. All chemicals in use should be disposed of in the appropriate waste containers and those waste containers removed from the lab. Once the natural radiocarbon laboratory has been thoroughly ''cleaned'', several background samples consisting of {sup 14}C depleted material should be processed through the lab and

  12. Multicenter validation of PCR-based method for detection of Salmonella in chicken and pig samples

    DEFF Research Database (Denmark)

    Malorny, B.; Cook, N.; D'Agostino, M.


    and pig swab samples. The 3 levels were 1-10, 10-100, and 100-1000 colony-forming units (CFU)/100 mL. Sample preparations, including inoculation and pre-enrichment in buffered peptone water (BPW), were performed centrally in a German laboratory; the pre-PCR sample preparation (by a resin-based method......As part of a standardization project, an interlaboratory trial including 15 laboratories from 13 European countries was conducted to evaluate the performance of a noproprietary polymerase chain reaction (PCR)-based method for the detection of Salmonella on artificially contaminated chicken rinse......) and PCR assay (gel electrophoresis detection) were performed by the receiving laboratories. Aliquots of BPW enrichment cultures were sent to the participants, who analyzed them using a thermal lysis procedure followed by a validated Salmonella-specific PCR assay. The results were reported as negative...

  13. Evaluation of neon focused ion beam milling for TEM sample preparation. (United States)

    Pekin, T C; Allen, F I; Minor, A M


    Gallium-based focused ion beams generated from liquid-metal sources are widely used in micromachining and sample preparation for transmission electron microscopy, with well-known drawbacks such as sample damage and contamination. In this work, an alternative (neon) focused ion beam generated by a gas field-ionization source is evaluated for the preparation of electron-transparent specimens. To do so, electron-transparent sections of Si and an Al alloy are prepared with both Ga and Ne ion beams for direct comparison. Diffraction-contrast imaging and energy dispersive x-ray spectroscopy are used to evaluate the relative damage induced by the two beams, and cross-sections of milled trenches are examined to compare the implantation depth with theoretical predictions from Monte Carlo simulations. Our results show that for the beam voltages and materials systems investigated, Ne ion beam milling does not significantly reduce the focused ion beam induced artefacts. However, the Ne ion beam does enable more precise milling and may be of interest in cases where Ga contamination cannot be tolerated. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  14. Preparation of Co2FeSn Heusler alloys by electrodeposition method

    Directory of Open Access Journals (Sweden)

    Nobuaki Watanabe


    Full Text Available We propose a new method for preparing Heusler alloys on the basis of the electrodeposition. The alloy composition is controllable by the deposition potential. The composition ratio of (Co, Fe and Sn is inversely proportional to the deposition potential. The effect of deposition potential on alloy composition, surface morphology, crystal structure, and magnetic properties of the samples was investigated. The (Co, Fe vs. Sn alloy composition displayed an inverse dependence on the deposition potential. According to the magnetic measurement, the obtained Co2FeSn composition alloy showed ferromagnetic properties. The coercivity shows the minimum value in the stoichiometric samples.

  15. Automated column liquid chromatographic determination of amoxicillin and cefadroxil in bovine serum and muscle tissue using on-line dialysis for sample preparation

    NARCIS (Netherlands)

    Snippe, N; van de Merbel, N C; Ruiter, F P; Steijger, O M; Lingeman, H; Brinkman, U A


    A fully automated method is described for the determination of amoxicillin and cefadroxil in bovine serum and muscle tissue. The method is based on the on-line combination of dialysis and solid-phase extraction for sample preparation, and column liquid chromatography with ultraviolet detection. In

  16. FC and ZFC magnetic properties of ferro-spinels (MFe2O4) prepared by solution-combustion method (United States)

    Aravind, G.; Kumar, R. Vijaya; Nathaniyal, V.; Rambabu, T.; Ravinder, D.


    Magnetic ferro-spinels MFe2O4 (M= Co and Ni) prepared by citrate-gel solution combustion method using metal nitrates with low sintering temperature (500°C). From the XRD and TEM studies confirm that a nano crystalline nature of the prepared samples. Field Cooled (FC) and Zero Field Cooled (ZFC) magnetic studies of the prepared ferro-spinels are measured by using vibrating sample magnetometer (VSM). The resultant magnetization of the prepared samples as a function of an applied magnetic field 10 T was measured at two different temperatures 5 K and 310 K. Field Cooled (FC) and Zero Field Cooled (ZFC) magnetization measurements under an applied field of 100 Oe and 1000 Oe in the temperature range of 5-375 K were carried out, which shows the blocking temperature of these two samples at around 350 K.

  17. Poly (lactic-co-glycolic acid) particles prepared by microfluidics and conventional methods. Modulated particle size and rheology. (United States)

    Perez, Aurora; Hernández, Rebeca; Velasco, Diego; Voicu, Dan; Mijangos, Carmen


    Microfluidic techniques are expected to provide narrower particle size distribution than conventional methods for the preparation of poly (lactic-co-glycolic acid) (PLGA) microparticles. Besides, it is hypothesized that the particle size distribution of poly (lactic-co-glycolic acid) microparticles influences the settling behavior and rheological properties of its aqueous dispersions. For the preparation of PLGA particles, two different methods, microfluidic and conventional oil-in-water emulsification methods were employed. The particle size and particle size distribution of PLGA particles prepared by microfluidics were studied as a function of the flow rate of the organic phase while particles prepared by conventional methods were studied as a function of stirring rate. In order to study the stability and structural organization of colloidal dispersions, settling experiments and oscillatory rheological measurements were carried out on aqueous dispersions of PLGA particles with different particle size distributions. Microfluidics technique allowed the control of size and size distribution of the droplets formed in the process of emulsification. This resulted in a narrower particle size distribution for samples prepared by MF with respect to samples prepared by conventional methods. Polydisperse samples showed a larger tendency to aggregate, thus confirming the advantages of microfluidics over conventional methods, especially if biomedical applications are envisaged. Copyright © 2014 Elsevier Inc. All rights reserved.

  18. Reproducibility of NMR analysis of urine samples: impact of sample preparation, storage conditions, and animal health status. (United States)

    Schreier, Christina; Kremer, Werner; Huber, Fritz; Neumann, Sindy; Pagel, Philipp; Lienemann, Kai; Pestel, Sabine


    Spectroscopic analysis of urine samples from laboratory animals can be used to predict the efficacy and side effects of drugs. This employs methods combining (1)H NMR spectroscopy with quantification of biomarkers or with multivariate data analysis. The most critical steps in data evaluation are analytical reproducibility of NMR data (collection, storage, and processing) and the health status of the animals, which may influence urine pH and osmolarity. We treated rats with a solvent, a diuretic, or a nephrotoxicant and collected urine samples. Samples were titrated to pH 3 to 9, or salt concentrations increased up to 20-fold. The effects of storage conditions and freeze-thaw cycles were monitored. Selected metabolites and multivariate data analysis were evaluated after (1)H NMR spectroscopy. We showed that variation of pH from 3 to 9 and increases in osmolarity up to 6-fold had no effect on the quantification of the metabolites or on multivariate data analysis. Storage led to changes after 14 days at 4°C or after 12 months at -20°C, independent of sample composition. Multiple freeze-thaw cycles did not affect data analysis. Reproducibility of NMR measurements is not dependent on sample composition under physiological or pathological conditions.

  19. Preparative chromatography for specific δ13C isotopic analysis of individual carbohydrates in environmental samples (United States)

    Nouara, Amel; Panagiotopoulos, Christos; Balesdent, Jérôme; Sempéré, Richard


    Carbohydrates are among the most abundant organic molecules on the Earth and are present in all geochemical systems. Despite their high abundance in the environment, very few studies assessed their origin using molecular carbohydrate isotopic analyses. In contrast with bulk stable isotope analysis (BSIA), which gives the isotopic signature of the entire sample without any specification about its chemical composition, compound specific 13C isotopic analysis of individual sugars (CSIA) offers valuable information about the origin of single molecules. Previous investigations used gas or liquid chromatography coupled with isotope ratio mass spectroscopy (GC-IRMS; HPLC-IRMS) for CSIA of sugars however the former requires δ13C corrections due to the carbon added to the sugar (derivatization) while the later does not provide always adequate separations among monosaccharides. Here we used cation preparative chromatography (Ca2+, Pb2+ and Na+) with refractive index detection in order to produce pure monosaccharide targets for subsequent EA-IRMS analyses. Milli-Q water was used as eluant at a flow rate 0.6 ml min-1. In general, three successive purifications (Ca2+, Pb2+, Ca2+) were sufficient to produce pure compounds. Pure monosaccharides were compared with authentic monosaccharide standards using 1H NMR and/or mass spectroscopy. The detection limit of our technique was about 1µM/sugar with a precision of 10% (n=6). Blanks run with Milli-Q water after three successive purifications resulted in carbon content of 0.13 to 2.77 µgC per collected sugar. These values are much lower than the minimum required amount (5 µgC) of the EA-IRSMS system with a precision of ± 0.35 ‰. Application of our method to environmental samples resulted in δ13C values of glucose, fructose, and levoglucosan in the range of -24 to -26 ‰ (PM10 atmospheric particles), and -15‰ to -22 ‰ for arabinose, glucose, and xylose (marine high molecular dissolved organic matter). These results fall in

  20. Liquid-solid sample preparation followed by headspace solid-phase microextraction determination of multiclass pesticides in soil. (United States)

    Durović, Rada D; Dordević, Tijana M; Santrić, Ljiljana R


    This paper describes development and validation of a multiresidue method for the determination of five pesticides (terbufos, prochloraz, chloridazon, pendimethalin, and fluorochloridone) belonging to different pesticide groups in soil samples by GC/MS, followed by its application in the analysis of some agricultural soil samples. The method is based on a headspace solid-phase microextraction method. Microextraction conditions, namely temperature, extraction time, and NaCI content, were tested and optimized using a 100 microm polydimethylsiloxane fiber. Three extraction solvents [methanol, methanol-acetone (1 + 1, v/v), and methanol-acetone-hexane (2 + 2 + 1, v/v/v)] and the optimum number of extraction steps within the sample preparation stage were optimized for the extraction procedure. LOD values for all the studied compounds were less than 12 microg/kg. Recovery values for multiple analyses of soil samples fortified at 30 microg/kg of each pesticide were higher than 64%. The method was proven to be repeatable, with RSD lower than 15%.

  1. System and method for measuring fluorescence of a sample

    Energy Technology Data Exchange (ETDEWEB)

    Riot, Vincent J.


    The present disclosure provides a system and a method for measuring fluorescence of a sample. The sample may be a polymerase-chain-reaction (PCR) array, a loop-mediated-isothermal amplification array, etc. LEDs are used to excite the sample, and a photodiode is used to collect the sample's fluorescence. An electronic offset signal is used to reduce the effects of background fluorescence and the noises from the measurement system. An integrator integrates the difference between the output of the photodiode and the electronic offset signal over a given period of time. The resulting integral is then converted into digital domain for further processing and storage.

  2. Soil separator and sampler and method of sampling (United States)

    O'Brien, Barry H [Idaho Falls, ID; Ritter, Paul D [Idaho Falls, ID


    A soil sampler includes a fluidized bed for receiving a soil sample. The fluidized bed may be in communication with a vacuum for drawing air through the fluidized bed and suspending particulate matter of the soil sample in the air. In a method of sampling, the air may be drawn across a filter, separating the particulate matter. Optionally, a baffle or a cyclone may be included within the fluidized bed for disentrainment, or dedusting, so only the finest particulate matter, including asbestos, will be trapped on the filter. The filter may be removable, and may be tested to determine the content of asbestos and other hazardous particulate matter in the soil sample.

  3. System and method for measuring fluorescence of a sample (United States)

    Riot, Vincent J


    The present disclosure provides a system and a method for measuring fluorescence of a sample. The sample may be a polymerase-chain-reaction (PCR) array, a loop-mediated-isothermal amplification array, etc. LEDs are used to excite the sample, and a photodiode is used to collect the sample's fluorescence. An electronic offset signal is used to reduce the effects of background fluorescence and the noises from the measurement system. An integrator integrates the difference between the output of the photodiode and the electronic offset signal over a given period of time. The resulting integral is then converted into digital domain for further processing and storage.

  4. Sperm preparation: state-of-the-art--physiological aspects and application of advanced sperm preparation methods. (United States)

    Henkel, Ralf


    For assisted reproduction technologies (ART), numerous techniques were developed to isolate spermatozoa capable of fertilizing oocytes. While early methodologies only focused on isolating viable, motile spermatozoa, with progress of ART, particularly intracytoplasmic sperm injection (ICSI), it became clear that these parameters are insufficient for the identification of the most suitable spermatozoon for fertilization. Conventional sperm preparation techniques, namely, swim-up, density gradient centrifugation and glass wool filtration, are not efficient enough to produce sperm populations free of DNA damage, because these techniques are not physiological and not modeled on the stringent sperm selection processes taking place in the female genital tract. These processes only allow one male germ cell out of tens of millions to fuse with the oocyte. Sites of sperm selection in the female genital tract are the cervix, uterus, uterotubal junction, oviduct, cumulus oophorus and the zona pellucida. Newer strategies of sperm preparation are founded on: (i) morphological assessment by means of 'motile sperm organelle morphological examination (MSOME)'; (ii) electrical charge; and (iii) molecular binding characteristics of the sperm cell. Whereas separation methods based on electrical charge take advantage of the sperm's adherence to a test tube surface or separate in an electrophoresis, molecular binding techniques use Annexin V or hyaluronic acid (HA) as substrates. Techniques in this category are magnet-activated cell sorting, Annexin V-activated glass wool filtration, flow cytometry and picked spermatozoa for ICSI (PICSI) from HA-coated dishes and HA-containing media. Future developments may include Raman microspectrometry, confocal light absorption and scattering spectroscopic microscopy and polarization microscopy.

  5. Using re-sampling methods in mortality studies.

    Directory of Open Access Journals (Sweden)

    Igor Itskovich

    Full Text Available Traditional methods of computing standardized mortality ratios (SMR in mortality studies rely upon a number of conventional statistical propositions to estimate confidence intervals for obtained values. Those propositions include a common but arbitrary choice of the confidence level and the assumption that observed number of deaths in the test sample is a purely random quantity. The latter assumption may not be fully justified for a series of periodic "overlapping" studies. We propose a new approach to evaluating the SMR, along with its confidence interval, based on a simple re-sampling technique. The proposed method is most straightforward and requires neither the use of above assumptions nor any rigorous technique, employed by modern re-sampling theory, for selection of a sample set. Instead, we include all possible samples that correspond to the specified time window of the study in the re-sampling analysis. As a result, directly obtained confidence intervals for repeated overlapping studies may be tighter than those yielded by conventional methods. The proposed method is illustrated by evaluating mortality due to a hypothetical risk factor in a life insurance cohort. With this method used, the SMR values can be forecast more precisely than when using the traditional approach. As a result, the appropriate risk assessment would have smaller uncertainties.

  6. Method for fractional solid-waste sampling and chemical analysis

    DEFF Research Database (Denmark)

    Riber, Christian; Rodushkin, I.; Spliid, Henrik


    Chemical characterization of solid waste is a demanding task due to the heterogeneity of the waste. This article describes how 45 material fractions hand-sorted from Danish household waste were subsampled and prepared for chemical analysis of 61 substances. All material fractions were subject...... of variance (20-85% of the overall variation). Only by increasing the sample size significantly can this variance be reduced. The accuracy and short-term reproducibility of the chemical characterization were good, as determined by the analysis of several relevant certified reference materials. Typically, six...

  7. Heat-capacity measurements on small samples: The hybrid method

    NARCIS (Netherlands)

    Klaasse, J.C.P.; Brück, E.H.


    A newly developed method is presented for measuring heat capacities on small samples, particularly where thermal isolation is not sufficient for the use of the traditional semiadiabatic heat-pulse technique. This "hybrid technique" is a modification of this heat-pulse method in case the temperature

  8. Method of determining an electrical property of a test sample

    DEFF Research Database (Denmark)


    A method of obtaining an electrical property of a test sample, comprising a non-conductive area and a conductive or semi-conductive test area, byperforming multiple measurements using a multi-point probe. The method comprising the steps of providing a magnetic field having field lines passing...... the electrical property of the test area....

  9. Size dependent photocatalytic activity of ZnS nanostructures prepared by a facile precipitation method

    Energy Technology Data Exchange (ETDEWEB)

    Yin, Lixiong, E-mail:; Zhang, Dongdong; Wang, Dan; Kong, Xingang; Huang, Jianfeng, E-mail:; Wang, Feifei; Wu, Yabo


    Highlights: • Size-controlled ZnS crystallites were achieved by adjusting the reaction pH. • The formation mechanism of ZnS prepared at different pH was discussed. • Size-related photocatalytic properties of ZnS were shown. - Abstract: Size-controlled ZnS nanostructures have been synthesized successfully with different reaction pH via a facile homogeneous precipitation method. These samples were characterized by X-ray diffraction, scanning electron microscopy (SEM), transmission electron microscopy (TEM), N{sub 2}-sorption BET surface area, Raman spectra and UV–vis spectroscopy for their structures, morphologies, specific surface area and band gaps values (E{sub g}). Results show that the size of the prepared nanospheres can be controllably tuned by adjusting pH value. Moreover, the size-related mechanism of different samples was also discussed. The degradation of methyl orange (MO) under UV light irradiation was carried out to study the effects of the size on the photocatalytic activity of the prepared samples. It was found that the sample obtained at pH = 7 exhibited the higher photocatalytic activity, which degradation efficiency reaches 99.5% under UV irradiation for 30 min. The big specific surface area of ZnS nanostructures are believed to greatly affect the final degradation efficiency of MO in our research. Moreover, the mechanism of enhanced photocatalytic activity was also investigated and the ·O{sup 2−} plays a dominant role during degradation process.



    Yasin Ünsal


    The aim of this study is to determine the basic problems encountered by counseling teachers in the fields of science and maths while preparing scientific projects. Screening model was used in the study. The data were collected by using survey method. Being developed for this purpose; the survey form was applied to 192 project counselors from various branches (like Physics, Chemistry, Biology, Maths, Science and Technology) at public schools in Turkey. As a result of the study, it was determin...

  11. Preparation of in-house reference soil sample containing high levels of naturally occurring radioactive materials from the oil industry. (United States)

    Al-Masri, M S; Aba, A; Al-Hamwi, A; Shakhashiro, A


    An in-house reference soil sample containing high levels of naturally occurring radioactive materials collected from contaminated areas in the Syrian oilfields has been prepared as a part of the quality assurance program in AECS. Homogeneity of the sample has been examined using three methods, viz. particle size distribution of the sample matrix, total alpha/beta counting and gamma spectrometry. In conjunction with Dixon and Grubb tests as statistical tools, ten random samples from the original sample were used for this investigation. Reference values for the three radium isotopes (224Ra, 226Ra, 228Ra) were determined using gamma spectrometry equipped with HPGe detectors having high relative efficiencies of 80%, while the reference value of 210Pb in the sample was determined using radiochemical separation and counting of its daughter 210Po by alpha spectrometry. ANOVA analysis was used to estimate the uncertainties due to measurement and inhomogeneity of the sample; uncertainty due to inhomogeneity was found to be around 2.6 times the measurement uncertainty.

  12. Efficiency of snake sampling methods in the Brazilian semiarid region. (United States)

    Mesquita, Paula C M D; Passos, Daniel C; Cechin, Sonia Z


    The choice of sampling methods is a crucial step in every field survey in herpetology. In countries where time and financial support are limited, the choice of the methods is critical. The methods used to sample snakes often lack objective criteria, and the traditional methods have apparently been more important when making the choice. Consequently researches using not-standardized methods are frequently found in the literature. We have compared four commonly used methods for sampling snake assemblages in a semiarid area in Brazil. We compared the efficacy of each method based on the cost-benefit regarding the number of individuals and species captured, time, and financial investment. We found that pitfall traps were the less effective method in all aspects that were evaluated and it was not complementary to the other methods in terms of abundance of species and assemblage structure. We conclude that methods can only be considered complementary if they are standardized to the objectives of the study. The use of pitfall traps in short-term surveys of the snake fauna in areas with shrubby vegetation and stony soil is not recommended.

  13. A multi-dimensional sampling method for locating small scatterers (United States)

    Song, Rencheng; Zhong, Yu; Chen, Xudong


    A multiple signal classification (MUSIC)-like multi-dimensional sampling method (MDSM) is introduced to locate small three-dimensional scatterers using electromagnetic waves. The indicator is built with the most stable part of signal subspace of the multi-static response matrix on a set of combinatorial sampling nodes inside the domain of interest. It has two main advantages compared to the conventional MUSIC methods. First, the MDSM is more robust against noise. Second, it can work with a single incidence even for multi-scatterers. Numerical simulations are presented to show the good performance of the proposed method.

  14. Advanced Markov chain Monte Carlo methods learning from past samples

    CERN Document Server

    Liang, Faming; Carrol, Raymond J


    This book provides comprehensive coverage of simulation of complex systems using Monte Carlo methods. Developing algorithms that are immune to the local trap problem has long been considered as the most important topic in MCMC research. Various advanced MCMC algorithms which address this problem have been developed include, the modified Gibbs sampler, the methods based on auxiliary variables and the methods making use of past samples. The focus of this book is on the algorithms that make use of past samples. This book includes the multicanonical algorithm, dynamic weighting, dynamically weight

  15. Phytotoxicity of Ag nanoparticles prepared by biogenic and chemical methods (United States)

    Choudhury, Rupasree; Majumder, Manna; Roy, Dijendra Nath; Basumallick, Srijita; Misra, Tarun Kumar


    Silver nanoparticles (Ag NPs) are now widely used as antibacterial and antifungal materials in different consumer products. We report here the preparation of Ag NPs by neem leaves extract ( Azadirachta) reduction and trisodium citrate-sodium borohydride reduction methods, and study of their phytotoxicity. The nanoparticles were characterized by UV-Vis spectroscopy, FTIR, and atomic force microscopy (AFM) techniques. Both neem-coated and citrate-coated Ag NPs exhibit surface plasmon around 400 nm, and their average sizes measured by AFM are about 100 and 20 nm, respectively. Antibacterial and antifungal activities of these nanomaterials have been studied by simple pea seed germination and disk diffusion methods. It has been observed from the growth of root and shoot, citrate-coated Ag NPs significantly affect seedling growth, but neem-coated Ag NPs exhibit somehow mild toxicity toward germination process due to the nutrient supplements from neem. On the other hand, antifungal activity of neem-coated Ag NPs has been found much higher than that of citrate-coated Ag NPs due to the combined effects of antifungal activity of neem and Ag NPs. Present research primarily indicates a possible application of neem-coated Ag NPs as a potential fungicide.

  16. Application of a Dual-Arm Robot in Complex Sample Preparation and Measurement Processes. (United States)

    Fleischer, Heidi; Drews, Robert Ralf; Janson, Jessica; Chinna Patlolla, Bharath Reddy; Chu, Xianghua; Klos, Michael; Thurow, Kerstin


    Automation systems with applied robotics have already been established in industrial applications for many years. In the field of life sciences, a comparable high level of automation can be found in the areas of bioscreening and high-throughput screening. Strong deficits still exist in the development of flexible and universal fully automated systems in the field of analytical measurement. Reasons are the heterogeneous processes with complex structures, which include sample preparation and transport, analytical measurements using complex sensor systems, and suitable data analysis and evaluation. Furthermore, the use of nonstandard sample vessels with various shapes and volumes results in an increased complexity. The direct use of existing automation solutions from bioscreening applications is not possible. A flexible automation system for sample preparation, analysis, and data evaluation is presented in this article. It is applied for the determination of cholesterol in biliary endoprosthesis using gas chromatography-mass spectrometry (GC-MS). A dual-arm robot performs both transport and active manipulation tasks to ensure human-like operation. This general robotic concept also enables the use of manual laboratory devices and equipment and is thus suitable in areas with a high standardization grade. © 2016 Society for Laboratory Automation and Screening.

  17. Preparation and Characterization of Celecoxib Dispersions in Soluplus®: Comparison of Spray Drying and Conventional Methods (United States)

    Homayouni, Alireza; Sadeghi, Fatemeh; Nokhodchi, Ali; Varshosaz, Jaleh; Afrasiabi Garekani, Hadi


    The present study deals with characterization of dispersions of a poorly water-soluble drug, celecoxib (CLX) in polyvinyl caprolactame–polyvinyl acetate–polyethylene glycol graft copolymer (Soluplus® (SOL)) prepared by different techniques. Dispersions of CLX in SOL at different ratios (2:1, 1:1, 1:2, 1:4 and 1:6) were prepared by spray drying, conventional solvent evaporation and melting methods. The solid states of samples were characterized using particle size measurements, optical and scanning electron microscopy, XRPD, DSC and FT-IR. The Gordon-Taylor equation was used to predict the Tg of samples and the possibility of interaction between CLX and SOL. The solubility and dissolution rate of all samples were determined. Stability of samples was studied at ambient conditions for a period of 12 months. DSC and XRPD analyses confirmed amorphous state of drug in samples. Surprisingly dispersions of CLX:SOL with the ratio of 2:1 and 1:1 showed slower dissolution rate than CLX while other samples showed higher dissolution rate. At 1:2 ratio the spray dried samples exhibited higher dissolution rate than corresponding samples prepared by other methods. However at higher SOL content (1:4 and 1:6), samples prepared by different methods showed similar dissolution profiles. The stability studies showed that there were no remarkable changes in the dissolution profiles and solid state of the drug after 12 months storage at ambient conditions. It was concluded that SOL was a proper carrier to enhance the dissolution rate of CLX. At high SOL ratios the method of preparation of dispersed samples had no effect on dissolution rate, whilst at low SOL content spray drying was more efficient method. PMID:25561910

  18. Sample size formulae for the Bayesian continual reassessment method. (United States)

    Cheung, Ying Kuen


    In the planning of a dose finding study, a primary design objective is to maintain high accuracy in terms of the probability of selecting the maximum tolerated dose. While numerous dose finding methods have been proposed in the literature, concrete guidance on sample size determination is lacking. With a motivation to provide quick and easy calculations during trial planning, we present closed form formulae for sample size determination associated with the use of the Bayesian continual reassessment method (CRM). We examine the sampling distribution of a nonparametric optimal design and exploit it as a proxy to empirically derive an accuracy index of the CRM using linear regression. We apply the formulae to determine the sample size of a phase I trial of PTEN-long in pancreatic cancer patients and demonstrate that the formulae give results very similar to simulation. The formulae are implemented by an R function 'getn' in the package 'dfcrm'. The results are developed for the Bayesian CRM and should be validated by simulation when used for other dose finding methods. The analytical formulae we propose give quick and accurate approximation of the required sample size for the CRM. The approach used to derive the formulae can be applied to obtain sample size formulae for other dose finding methods.

  19. Thermoluminescence study of aluminium oxide doped germanium prepared by combustion synthesis method

    Directory of Open Access Journals (Sweden)

    Saharin Nurul Syazlin Binti


    Full Text Available The present paper reports the optimum concentration of germanium (Ge dopant in aluminium oxide (AhO3 samples prepared by combustion synthesis (CS method for thermoluminescence (TL studies. The samples were prepared at various Ge concentration i.e. 1 to 5% mol. The phase formation of un-doped and Ge-doped Al2O3 samples was determined using X-ray Diffraction (XRD. The sharp peaks present in the XRD pattern confirms the crystallinity of the samples. The samples were then exposed to 50 Gy Cobalt-60 sources (Gamma cell 220. TL glow curves were measured and recorded using a Harshaw Model 3500 TLD reader. Comparison of TL peaks were observed to obtain the best composition of Ge dopants. A simple glow curves TL peak at around 175̊C for all composition samples was observed. It was also found that the composition of aluminium oxide doped with 3.0% of germanium exhibits the highest thermoluminescence (TL intensity which is 349747.04 (a.u.

  20. Differentiation and classification of beers with flame atomic spectrometry and molecular absorption spectrometry and sample preparation assisted by microwaves (United States)

    Bellido-Milla, Dolores; Moreno-Perez, Juana M.; Hernández-Artiga, María. P.


    The characterization of beer samples has a lot of interest because their composition can affect the taste and stability of beer and consumer health. Flame atomic absorption spectrometry was used to determine Fe, Mn, Zn, Cu, Mg, Ca and Al. Sodium and K were determined by flame atomic emission spectrometry. A sample preparation method was developed, based on treatment with HNO 3 and H 2O 2 in a microwave oven. This has many advantages over the methods found in the literature. The combination of the results of atomic spectrometry and the spectrum obtained by molecular absorption spectrometry provides information on the inorganic and organic components of the samples. The application of chemometric techniques to chemical composition data could be extremely useful for food quality control. The metal concentrations, the molecular absorption spectrum, the pH and conductivity of each sample were subject to analysis of variance and linear discriminant analysis. Twenty-five different beer samples were used to differentiate and classify different types of beers.

  1. High Throughput Sample Preparation and Analysis for DNA Sequencing, PCR and Combinatorial Screening of Catalysis Based on Capillary Array Technique

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yonghua [Iowa State Univ., Ames, IA (United States)


    Sample preparation has been one of the major bottlenecks for many high throughput analyses. The purpose of this research was to develop new sample preparation and integration approach for DNA sequencing, PCR based DNA analysis and combinatorial screening of homogeneous catalysis based on multiplexed capillary electrophoresis with laser induced fluorescence or imaging UV absorption detection. The author first introduced a method to integrate the front-end tasks to DNA capillary-array sequencers. protocols for directly sequencing the plasmids from a single bacterial colony in fused-silica capillaries were developed. After the colony was picked, lysis was accomplished in situ in the plastic sample tube using either a thermocycler or heating block. Upon heating, the plasmids were released while chromsomal DNA and membrane proteins were denatured and precipitated to the bottom of the tube. After adding enzyme and Sanger reagents, the resulting solution was aspirated into the reaction capillaries by a syringe pump, and cycle sequencing was initiated. No deleterious effect upon the reaction efficiency, the on-line purification system, or the capillary electrophoresis separation was observed, even though the crude lysate was used as the template. Multiplexed on-line DNA sequencing data from 8 parallel channels allowed base calling up to 620 bp with an accuracy of 98%. The entire system can be automatically regenerated for repeated operation. For PCR based DNA analysis, they demonstrated that capillary electrophoresis with UV detection can be used for DNA analysis starting from clinical sample without purification. After PCR reaction using cheek cell, blood or HIV-1 gag DNA, the reaction mixtures was injected into the capillary either on-line or off-line by base stacking. The protocol was also applied to capillary array electrophoresis. The use of cheaper detection, and the elimination of purification of DNA sample before or after PCR reaction, will make this approach an

  2. Preparation of Al/Si functionally graded materials using ultrasonic separation method

    Directory of Open Access Journals (Sweden)

    Zhang Zhongtao


    Full Text Available Functionally graded materials (FGM have been widely used in many industries such as aerospace, energy and electronics. In this experimental study of fabricating FGM, an approach was developed to prepare Al/Si FGM using power ultrasonic separation method. Material sample with continuously changing composition and performance/properties was successfully produced. Results showed that the microstructure of the FGM sample transited, from its top to bottom, from the hypereutectic structure with a large quantity of primary Si gradually to the eutectic, and fi nally to the hypoeutectic with numerous primary Al dendrites. The distribution of primary Si and microhardness of the FGM sample also presented graded characteristics, resulting that the wear resistance of the FGM sample decreased from top to bottom. Preliminary discussion was made on the mechanism of the formation of Al/Si FGM.

  3. Novel structural and magnetic properties of Mg doped copper nanoferrites prepared by conventional and wet methods (United States)

    Ahmed, M. A.; Afify, H. H.; El Zawawia, I. K.; Azab, A. A.


    Nanoferrites of the general formula Cu1-xMgxFe2O4 with 0≤x≤0.6 were prepared by standard ceramic and wet methods. The structure was studied by X-ray diffraction and IR spectroscopy. The density and lattice constant were calculated and reported. The particle size of the prepared nanoferrites ranged from 8.7 to 41.1 nm. It was found that the lattice parameter decreases with increasing cation substitution of Mg2+ due to the difference of ionic radius and atomic mass. The dc magnetic susceptibility was measured out using Faraday's method. The magnetic hysteresis measurement was performed using a vibrating sample magnetometer. Magnetic constants such as Curie temperature, effective magnetic moment, saturation magnetization, remanent magnetization and corecivicty were obtained and reported. The magnetic constants decrease with increasing Mg2+, except the remanent magnetization which increased.

  4. Sample preparation with an automated robotic workstation for organic acid analysis by gas chromatography-mass spectrometry. (United States)

    Bengtsson, I M; Lehotay, D C


    We attempted to automate sample preparation for analysis of organic acids by gas chromatography-mass spectrometry using a computer-controlled, automated robotic workstation that is integrated and connected to the gas chromatography-mass spectrometry (HP-5890/5971) system. Of the two methods developed, one employed solvent extraction, while the other utilized a silica, solid-phase extraction cartridge. Both automated methods were compared to a manual, solvent extraction procedure used routinely in our laboratory. Normal, spiked urine, and urine from patients with a variety of metabolic abnormalities were analyzed. The robotic workstation did not meet all our requirements for a rapid, reliable, laboratory device. Recoveries with the automated procedures were less than with the manual method, and some organic acids important in the diagnosis of inborn errors of metabolism were not detected. Additionally, the robotic device had mechanical and design problems that made it slower and less reliable than the manual procedure.


    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, S.; Culligan, B.


    A new rapid separation method for radiostrontium in emergency milk samples was developed at the Savannah River Site (SRS) Environmental Bioassay Laboratory (Aiken, SC, USA) that will allow rapid separation and measurement of Sr-90 within 8 hours. The new method uses calcium phosphate precipitation, nitric acid dissolution of the precipitate to coagulate residual fat/proteins and a rapid strontium separation using Sr Resin (Eichrom Technologies, Darien, IL, USA) with vacuum-assisted flow rates. The method is much faster than previous method that use calcination or cation exchange pretreatment, has excellent chemical recovery, and effectively removes beta interferences. When a 100 ml sample aliquot is used, the method has a detection limit of 0.5 Bq/L, well below generic emergency action levels.

  6. Preparation and characterization of ZnS nanoparticles prepared by hydrothermal method (United States)

    Wang, Caifeng; Li, Qingshan; Hu, Bo


    ZnS nanoparticles were prepared by hydrothermal method using zinc nitrate [Zn(NO3)2 ṡ 6H2O] and thiourea [SC(NH2)2] as sources of Zn2+ and S2- ions and cetyltrimethyl ammonium bromide [CH3(CH2)15N+(CH3)3 ṡ Br-; CTAB] as a surface active agent. The structural, surface morphology and optical properties of ZnS nanoparticles as a function of growth temperature were investigated. The studies show that ZnS nanoparticles have high transmittance over 70% in the visible light range of 400-800 nm, and it has a strong green emission band at about 520 nm which originates from the recombination of electrons from the energy level of sulfur vacancies with the holes from the energy level of zinc vacancies. With the increase of the growth temperature, the XRD peak intensity and the size of ZnS nanoparticles increase, while the green emission intensity firstly increases and then decreases.

  7. A user-friendly robotic sample preparation program for fully automated biological sample pipetting and dilution to benefit the regulated bioanalysis. (United States)

    Jiang, Hao; Ouyang, Zheng; Zeng, Jianing; Yuan, Long; Zheng, Naiyu; Jemal, Mohammed; Arnold, Mark E


    Biological sample dilution is a rate-limiting step in bioanalytical sample preparation when the concentrations of samples are beyond standard curve ranges, especially when multiple dilution factors are needed in an analytical run. We have developed and validated a Microsoft Excel-based robotic sample preparation program (RSPP) that automatically transforms Watson worklist sample information (identification, sequence and dilution factor) to comma-separated value (CSV) files. The Freedom EVO liquid handler software imports and transforms the CSV files to executable worklists (.gwl files), allowing the robot to perform sample dilutions at variable dilution factors. The dynamic dilution range is 1- to 1000-fold and divided into three dilution steps: 1- to 10-, 11- to 100-, and 101- to 1000-fold. The whole process, including pipetting samples, diluting samples, and adding internal standard(s), is accomplished within 1 h for two racks of samples (96 samples/rack). This platform also supports online sample extraction (liquid-liquid extraction, solid-phase extraction, protein precipitation, etc.) using 96 multichannel arms. This fully automated and validated sample dilution and preparation process has been applied to several drug development programs. The results demonstrate that application of the RSPP for fully automated sample processing is efficient and rugged. The RSPP not only saved more than 50% of the time in sample pipetting and dilution but also reduced human errors. The generated bioanalytical data are accurate and precise; therefore, this application can be used in regulated bioanalysis.

  8. Characterizing lentic freshwater fish assemblages using multiple sampling methods. (United States)

    Fischer, Jesse R; Quist, Michael C


    Characterizing fish assemblages in lentic ecosystems is difficult, and multiple sampling methods are almost always necessary to gain reliable estimates of indices such as species richness. However, most research focused on lentic fish sampling methodology has targeted recreationally important species, and little to no information is available regarding the influence of multiple methods and timing (i.e., temporal variation) on characterizing entire fish assemblages. Therefore, six lakes and impoundments (48-1,557 ha surface area) were sampled seasonally with seven gear types to evaluate the combined influence of sampling methods and timing on the number of species and individuals sampled. Probabilities of detection for species indicated strong selectivities and seasonal trends that provide guidance on optimal seasons to use gears when targeting multiple species. The evaluation of species richness and number of individuals sampled using multiple gear combinations demonstrated that appreciable benefits over relatively few gears (e.g., to four) used in optimal seasons were not present. Specifically, over 90 % of the species encountered with all gear types and season combinations (N = 19) from six lakes and reservoirs were sampled with nighttime boat electrofishing in the fall and benthic trawling, modified-fyke, and mini-fyke netting during the summer. Our results indicated that the characterization of lentic fish assemblages was highly influenced by the selection of sampling gears and seasons, but did not appear to be influenced by waterbody type (i.e., natural lake, impoundment). The standardization of data collected with multiple methods and seasons to account for bias is imperative to monitoring of lentic ecosystems and will provide researchers with increased reliability in their interpretations and decisions made using information on lentic fish assemblages.

  9. Self-contained cryogenic gas sampling apparatus and method (United States)

    McManus, G.J.; Motes, B.G.; Bird, S.K.; Kotter, D.K.


    Apparatus for obtaining a whole gas sample, is composed of: a sample vessel having an inlet for receiving a gas sample; a controllable valve mounted for controllably opening and closing the inlet; a valve control coupled to the valve for opening and closing the valve at selected times; a portable power source connected for supplying operating power to the valve control; and a cryogenic coolant in thermal communication with the vessel for cooling the interior of the vessel to cryogenic temperatures. A method is described for obtaining an air sample using the apparatus described above, by: placing the apparatus at a location at which the sample is to be obtained; operating the valve control to open the valve at a selected time and close the valve at a selected subsequent time; and between the selected times maintaining the vessel at a cryogenic temperature by heat exchange with the coolant. 3 figs.

  10. A lab-on-a-chip system with integrated sample preparation and loop-mediated isothermal amplification for rapid and quantitative detection of Salmonella spp. in food samples. (United States)

    Sun, Yi; Quyen, Than Linh; Hung, Tran Quang; Chin, Wai Hoe; Wolff, Anders; Bang, Dang Duong


    Foodborne disease is a major public health threat worldwide. Salmonellosis, an infectious disease caused by Salmonella spp., is one of the most common foodborne diseases. Isolation and identification of Salmonella by conventional bacterial culture or molecular-based methods are time consuming and usually take a few hours to days to complete. In response to the demand for rapid on line or on site detection of pathogens, in this study, we describe for the first time an eight-chamber lab-on-a-chip (LOC) system with integrated magnetic bead-based sample preparation and loop-mediated isothermal amplification (LAMP) for rapid and quantitative detection of Salmonella spp. in food samples. The whole diagnostic procedures including DNA isolation, isothermal amplification, and real-time detection were accomplished in a single chamber. Up to eight samples could be handled simultaneously and the system was capable to detect Salmonella at concentration of 50 cells per test within 40 min. The simple design, together with high level of integration, isothermal amplification, and quantitative analysis of multiple samples in short time, will greatly enhance the practical applicability of the LOC system for rapid on-site screening of Salmonella for applications in food safety control, environmental surveillance, and clinical diagnostics.

  11. Preparative separation of sesamin and sesamolin from defatted sesame meal via centrifugal partition chromatography with consecutive sample injection. (United States)

    Jeon, Je-Seung; Park, Chae Lee; Syed, Ahmed Shah; Kim, Young-Mi; Cho, Il Je; Kim, Chul Young


    A preparative separation method using consecutive sample injection centrifugal partition chromatography (CPC) was developed to obtain sesamin and sesamolin from defatted sesame meal extracts. A two-phase solvent system consisting of n-hexane-ethyl acetate-methanol-water (8:2:8:2, v/v) was applied in reversed-phase mode (descending mode). Preliminary experiments with an SCPC-100 (column volume: 100mL) were performed to select the appropriate two-phase solvent system and sample injection times; these parameters were then used with an SCPC-1000 (column volume: 1000mL) in a 10-fold scale-up preparative run. A sample containing 3g of crude extract was consecutively injected four times onto the SCPC-1000, which yielded 328mg of sesamin and 168mg of sesamolin. These compounds were analyzed by high-performance liquid chromatography and determined to have purities of 95.6% and 93.9%, respectively. Sesamin and sesamolin (30μM) increased antioxidant response element (ARE) luciferase activity 2.6-fold and 1.9-fold, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Methods for Sampling and Measurement of Compressed Air Contaminants

    Energy Technology Data Exchange (ETDEWEB)

    Stroem, L.


    In order to improve the technique for measuring oil and water entrained in a compressed air stream, a laboratory study has been made of some methods for sampling and measurement. For this purpose water or oil as artificial contaminants were injected in thin streams into a test loop, carrying dry compressed air. Sampling was performed in a vertical run, down-stream of the injection point. Wall attached liquid, coarse droplet flow, and fine droplet flow were sampled separately. The results were compared with two-phase flow theory and direct observation of liquid behaviour. In a study of sample transport through narrow tubes, it was observed that, below a certain liquid loading, the sample did not move, the liquid remaining stationary on the tubing wall. The basic analysis of the collected samples was made by gravimetric methods. Adsorption tubes were used with success to measure water vapour. A humidity meter with a sensor of the aluminium oxide type was found to be unreliable. Oil could be measured selectively by a flame ionization detector, the sample being pretreated in an evaporation- condensation unit

  13. Evaluation of research methods to study domestic food preparation

    NARCIS (Netherlands)

    Bongoni, R.; Verkerk, R.; Dekker, M.; Steenbekkers, B.


    Purpose – Domestic preparation practices influence the sensory properties and nutritional composition of food products. Information on the variability in actual domestic preparation practices is needed to assess the influence of applied conditions on the sensory and nutritional quality of food. The

  14. Polypyrrole solid phase microextraction: A new approach to rapid sample preparation for the monitoring of antibiotic drugs

    Energy Technology Data Exchange (ETDEWEB)

    Szultka, Malgorzata [Department of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus, Copernicus University, Gagarin 7, 87-100 Torun (Poland); Kegler, Ricarda [Institute of Clinical Pharmacology, University of Rostock, Schillingallee 70, D-18057 Rostock (Germany); Fuchs, Patricia [Department of Anaesthesia and Intensive Care, University of Rostock, Schillingallee 35, D-18057 Rostock (Germany); Olszowy, Pawel [Department of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus, Copernicus University, Gagarin 7, 87-100 Torun (Poland); Miekisch, Wolfram; Schubert, Jochen K. [Department of Anaesthesia and Intensive Care, University of Rostock, Schillingallee 35, D-18057 Rostock (Germany); Buszewski, Boguslaw [Department of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus, Copernicus University, Gagarin 7, 87-100 Torun (Poland); Mundkowski, Ralf G., E-mail: [Institute of Clinical Pharmacology, University of Rostock, Schillingallee 70, D-18057 Rostock (Germany)


    Simple or even rapid bioanalytical methods are rare, since they generally involve complicated, time-consuming sample preparation from the biological matrices like LLE or SPE. SPME provides a promising approach to overcome these limitations. The full potential of this innovative technique for medical diagnostics, pharmacotherapy or biochemistry has not been tapped yet. In-house manufactured SPME probes with polypyrrole (PPy) coating were evaluated using three antibiotics of high clinical relevance - linezolid, daptomycin, and moxifloxacin - from PBS, plasma, and whole blood. The PPy coating was characterised by scanning electron microscopy. Influences of pH, inorganic salt, and blood anticoagulants were studied for optimum performance. Extraction yields were determined from stagnant media as well as re-circulating human blood using the heart-and-lung machine model system. The PPy-SPME fibres showed high extraction yields, particularly regarding linezolid. The reproducibility of the method was optimised to achieve RSDs of 9% or 17% and 7% for SPME from stagnant or re-circulating blood using fresh and re-used fibres, respectively. The PPy-SPME approach was demonstrated to meet the requirements of therapeutic monitoring of the drugs tested, even from re-circulating blood at physiological flow rates. SPME represents a rapid and simple dual-step procedure with potency to significantly reduce the effort and expenditure of complicated sample preparations in biomedical analysis.

  15. Evaluating focused ion beam and ultramicrotome sample preparation for analytical microscopies of the cathode layer of a polymer electrolyte membrane fuel cell (United States)

    de A. Melo, Lis G.; Hitchcock, Adam P.; Berejnov, Viatcheslav; Susac, Darija; Stumper, Juergen; Botton, Gianluigi A.


    Optimizing the structure of the porous electrodes of polymer electrolyte membrane fuel cells (PEM-FC) can improve device power and durability. Analytical microscopy techniques are important tools for measuring the electrode structure, thereby providing guidance for structural optimization. Transmission Electron Microscopy (TEM), with either Energy Dispersive X-Ray (EDX) or Electron Energy Loss Spectroscopy (EELS) analysis, and Scanning Transmission X-Ray Microscopy (STXM) are complementary methods which, together, provide a powerful approach for PEM-FC electrode analysis. Both TEM and STXM require thin (50-200 nm) samples, which can be prepared either by Focused Ion Beam (FIB) milling or by embedding and ultramicrotomy. Here we compare TEM and STXM spectromicroscopy analysis of FIB and ultramicrotomy sample preparations of the same PEM-FC sample, with focus on how sample preparation affects the derived chemical composition and spatial distributions of carbon support and ionomer. The FIB lamella method, while avoiding pore-filling by embedding media, had significant problems. In particular, in the FIB sample the carbon support was extensively amorphized and the ionomer component suffered mass loss and structural damage. Although each sample preparation technique has a role to play in PEM-FC optimization studies, it is important to be aware of the limitations of each method.

  16. Antifungal activity of gold nanoparticles prepared by solvothermal method

    Energy Technology Data Exchange (ETDEWEB)

    Ahmad, Tokeer, E-mail: [Nanochemistry Laboratory, Department of Chemistry, Jamia Millia Islamia, New Delhi 110025 (India); Wani, Irshad A.; Lone, Irfan H.; Ganguly, Aparna [Nanochemistry Laboratory, Department of Chemistry, Jamia Millia Islamia, New Delhi 110025 (India); Manzoor, Nikhat; Ahmad, Aijaz [Department of Biosciences, Jamia Millia Islamia, New Delhi 110025 (India); Ahmed, Jahangeer [Department of Chemistry, Michigan State University, East Lansing, MI 48824 (United States); Al-Shihri, Ayed S. [Department of Chemistry, Faculty of Science, King Khalid University, Abha 61413, P.O. Box 9004 (Saudi Arabia)


    Graphical abstract: Gold nanoparticles (7 and 15 nm) of very high surface area (329 and 269 m{sup 2}/g) have been successfully synthesized through solvothermal method by using tin chloride and sodium borohydride as reducing agents. As-prepared gold nanoparticles shows very excellent antifungal activity against Candida isolates and activity increases with decrease in the particle size. Display Omitted Highlights: ► Effect of reducing agents on the morphology of gold nanoparticles. ► Highly uniform and monodisperse gold nanoparticles (7 nm). ► Highest surface area of gold nanoparticles (329 m{sup 2/}g). ► Excellent antifungal activity of gold nanoparticles against Candida strains. -- Abstract: Gold nanoparticles have been successfully synthesized by solvothermal method using SnCl{sub 2} and NaBH{sub 4} as reducing agents. X-ray diffraction studies show highly crystalline and monophasic nature of the gold nanoparticles with face centred cubic structure. The transmission electron microscopic studies show the formation of nearly spherical gold nanoparticles of average size of 15 nm using SnCl{sub 2}, however, NaBH{sub 4} produced highly uniform, monodispersed and spherical gold nanoparticles of average grain size of 7 nm. A high surface area of 329 m{sup 2}/g for 7 nm and 269 m{sup 2}/g for 15 nm gold nanoparticles was observed. UV–vis studies assert the excitations over the visible region due to transverse and longitudinal surface plasmon modes. The gold nanoparticles exhibit excellent size dependant antifungal activity and greater biocidal action against Candida isolates for 7 nm sized gold nanoparticles restricting the transmembrane H{sup +} efflux of the Candida species than 15 nm sized gold nanoparticles.

  17. Simplified platelet sample preparation for SDS-PAGE-based proteomic studies. (United States)

    Reicheltová, Zuzana; Májek, Pavel; Riedel, Tomáš; Suttnar, Jiří; Dyr, Jan E


    The goal of this study was to design an easy and simple protocol for platelet isolation and sample preparation for proteomic studies based on 2DE (IEF-SDS-PAGE) followed by Coomassie blue staining. Blood was collected by venipuncture into tubes coated with EDTA and platelet-rich plasma (PRP) was immediately obtained by centrifugation. PRP was stored refrigerated in closed Falcon tubes for 0, 1, 2, 3, 5, and 7 days and platelets were isolated by centrifugation. 2DE gels were stained with colloidal Coomassie blue stain and evaluated using the Progenesis SameSpots software. Spots that differed significantly in the gels of fresh and stored platelet samples were excised, digested with trypsin, and further analyzed using nanoLC-MS/MS. During the 7-day follow-up period, we found 20 spots that differed significantly (ANOVA p investigations, whenever it is not feasible to prepare washed platelets immediately after blood collection, the EDTA-anticoagulated PRP can be stored in test tubes at 4°C for up to 2 days for the platelet proteome investigation. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Microfluidics cell sample preparation for analysis: Advances in efficient cell enrichment and precise single cell capture. (United States)

    Huang, Liang; Bian, Shengtai; Cheng, Yinuo; Shi, Guanya; Liu, Peng; Ye, Xiongying; Wang, Wenhui


    Single cell analysis has received increasing attention recently in both academia and clinics, and there is an urgent need for effective upstream cell sample preparation. Two extremely challenging tasks in cell sample preparation-high-efficiency cell enrichment and precise single cell capture-have now entered into an era full of exciting technological advances, which are mostly enabled by microfluidics. In this review, we summarize the category of technologies that provide new solutions and creative insights into the two tasks of cell manipulation, with a focus on the latest development in the recent five years by highlighting the representative works. By doing so, we aim both to outline the framework and to showcase example applications of each task. In most cases for cell enrichment, we take circulating tumor cells (CTCs) as the target cells because of their research and clinical importance in cancer. For single cell capture, we review related technologies for many kinds of target cells because the technologies are supposed to be more universal to all cells rather than CTCs. Most of the mentioned technologies can be used for both cell enrichment and precise single cell capture. Each technology has its own advantages and specific challenges, which provide opportunities for researchers in their own area. Overall, these technologies have shown great promise and now evolve into real clinical applications.

  19. Monoaminergic uptake in synaptosomes prepared from frozen brain tissue samples of normal and narcoleptic canines. (United States)

    Valtier, D; Dement, W C; Mignot, E


    Canine narcolepsy, a model of the human disorder, is associated with altered catecholamine but not serotonin (5-HT) metabolism in some brain areas, particularly the amygdala. A possible explanation for these global changes could be the existence of specific defects in monoamine uptake processes. We have studied the uptake of [3H]norepinephrine (NE), [3H]dopamine (DA) and [3H]5-HT in synaptosomes prepared from cortex and amygdala of narcoleptic and control Doberman pinscher brains. Since narcoleptic canines are relatively few in number, we have used a specific brain freezing procedure that has been reported to allow restoration of metabolically functional tissue upon thawing. Preliminary studies comparing monoamine uptake in fresh and frozen brain samples of both groups of dogs were carried out and demonstrated that this procedure significantly altered serotoninergic but not noradrenergic and dopaminergic uptake. All further investigations were then done on synaptosomes prepared from frozen samples. Our results demonstrate that synaptosomal uptake of [3H]NE, [3H]DA and [3H]5-HT in cortex and amygdala are not altered in narcolepsy.

  20. Review of online coupling of sample preparation techniques with liquid chromatography. (United States)

    Pan, Jialiang; Zhang, Chengjiang; Zhang, Zhuomin; Li, Gongke


    Sample preparation is still considered as the bottleneck of the whole analytical procedure, and efforts has been conducted towards the automation, improvement of sensitivity and accuracy, and low comsuption of organic solvents. Development of online sample preparation techniques (SP) coupled with liquid chromatography (LC) is a promising way to achieve these goals, which has attracted great attention. This article reviews the recent advances on the online SP-LC techniques. Various online SP techniques have been described and summarized, including solid-phase-based extraction, liquid-phase-based extraction assisted with membrane, microwave assisted extraction, ultrasonic assisted extraction, accelerated solvent extraction and supercritical fluids extraction. Specially, the coupling approaches of online SP-LC systems and the corresponding interfaces have been discussed and reviewed in detail, such as online injector, autosampler combined with transport unit, desorption chamber and column switching. Typical applications of the online SP-LC techniques have been summarized. Then the problems and expected trends in this field are attempted to be discussed and proposed in order to encourage the further development of online SP-LC techniques. Copyright © 2014 Elsevier B.V. All rights reserved.