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Sample records for scale liquid chromatography

  1. Towards Chip Scale Liquid Chromatography and High Throughput Immunosensing

    Energy Technology Data Exchange (ETDEWEB)

    Ni, Jing [Iowa State Univ., Ames, IA (United States)

    2000-09-21

    This work describes several research projects aimed towards developing new instruments and novel methods for high throughput chemical and biological analysis. Approaches are taken in two directions. The first direction takes advantage of well-established semiconductor fabrication techniques and applies them to miniaturize instruments that are workhorses in analytical laboratories. Specifically, the first part of this work focused on the development of micropumps and microvalves for controlled fluid delivery. The mechanism of these micropumps and microvalves relies on the electrochemically-induced surface tension change at a mercury/electrolyte interface. A miniaturized flow injection analysis device was integrated and flow injection analyses were demonstrated. In the second part of this work, microfluidic chips were also designed, fabricated, and tested. Separations of two fluorescent dyes were demonstrated in microfabricated channels, based on an open-tubular liquid chromatography (OT LC) or an electrochemically-modulated liquid chromatography (EMLC) format. A reduction in instrument size can potentially increase analysis speed, and allow exceedingly small amounts of sample to be analyzed under diverse separation conditions. The second direction explores the surface enhanced Raman spectroscopy (SERS) as a signal transduction method for immunoassay analysis. It takes advantage of the improved detection sensitivity as a result of surface enhancement on colloidal gold, the narrow width of Raman band, and the stability of Raman scattering signals to distinguish several different species simultaneously without exploiting spatially-separated addresses on a biochip. By labeling gold nanoparticles with different Raman reporters in conjunction with different detection antibodies, a simultaneous detection of a dual-analyte immunoassay was demonstrated. Using this scheme for quantitative analysis was also studied and preliminary dose-response curves from an immunoassay of a

  2. Optimization of large-scale pseudotargeted metabolomics method based on liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Luo, Ping; Yin, Peiyuan; Zhang, Weijian; Zhou, Lina; Lu, Xin; Lin, Xiaohui; Xu, Guowang

    2016-03-11

    Liquid chromatography-mass spectrometry (LC-MS) is now a main stream technique for large-scale metabolic phenotyping to obtain a better understanding of genomic functions. However, repeatability is still an essential issue for the LC-MS based methods, and convincing strategies for long time analysis are urgently required. Our former reported pseudotargeted method which combines nontargeted and targeted analyses, is proved to be a practical approach with high-quality and information-rich data. In this study, we developed a comprehensive strategy based on the pseudotargeted analysis by integrating blank-wash, pooled quality control (QC) sample, and post-calibration for the large-scale metabolomics study. The performance of strategy was optimized from both pre- and post-acquisition sections including the selection of QC samples, insertion frequency of QC samples, and post-calibration methods. These results imply that the pseudotargeted method is rather stable and suitable for large-scale study of metabolic profiling. As a proof of concept, the proposed strategy was applied to the combination of 3 independent batches within a time span of 5 weeks, and generated about 54% of the features with coefficient of variations (CV) below 15%. Moreover, the stability and maximal capability of a single analytical batch could be extended to at least 282 injections (about 110h) while still providing excellent stability, the CV of 63% metabolic features was less than 15%. Taken together, the improved repeatability of our strategy provides a reliable protocol for large-scale metabolomics studies. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Unifying expression scale for peptide hydrophobicity in proteomic reversed phase high-pressure liquid chromatography experiments.

    Science.gov (United States)

    Grigoryan, Marine; Shamshurin, Dmitry; Spicer, Victor; Krokhin, Oleg V

    2013-11-19

    As an initial step in our efforts to unify the expression of peptide retention times in proteomic liquid chromatography-mass spectrometry (LC-MS) experiments, we aligned the chromatographic properties of a number of peptide retention standards against a collection of peptides commonly observed in proteomic experiments. The standard peptide mixtures and tryptic digests of samples of different origins were separated under the identical chromatographic condition most commonly employed in proteomics: 100 Å C18 sorbent with 0.1% formic acid as an ion-pairing modifier. Following our original approach (Krokhin, O. V.; Spicer, V. Anal. Chem. 2009, 81, 9522-9530) the retention characteristics of these standards and collection of tryptic peptides were mapped into hydrophobicity index (HI) or acetonitrile percentage units. This scale allows for direct visualization of the chromatographic outcome of LC-MS acquisitions, monitors the performance of the gradient LC system, and simplifies method development and interlaboratory data alignment. Wide adoption of this approach would significantly aid understanding the basic principles of gradient peptide RP-HPLC and solidify our collective efforts in acquiring confident peptide retention libraries, a key component in the development of targeted proteomic approaches.

  4. performance liquid chromatography

    African Journals Online (AJOL)

    user

    2010-11-22

    Nov 22, 2010 ... This study is aimed to determine the concentration of acrylamide in processed food products available in open market. In order to determine the acrylamide concentration, three bakery items and three fried chips from three different brands were analyzed. High-performance liquid chromatography (HPLC) ...

  5. High Performance Liquid Chromatography

    Science.gov (United States)

    Talcott, Stephen

    High performance liquid chromatography (HPLC) has many applications in food chemistry. Food components that have been analyzed with HPLC include organic acids, vitamins, amino acids, sugars, nitrosamines, certain pesticides, metabolites, fatty acids, aflatoxins, pigments, and certain food additives. Unlike gas chromatography, it is not necessary for the compound being analyzed to be volatile. It is necessary, however, for the compounds to have some solubility in the mobile phase. It is important that the solubilized samples for injection be free from all particulate matter, so centrifugation and filtration are common procedures. Also, solid-phase extraction is used commonly in sample preparation to remove interfering compounds from the sample matrix prior to HPLC analysis.

  6. Untargeted large-scale plant metabolomics using liquid chromatography coupled to mass spectrometry

    NARCIS (Netherlands)

    Vos, de C.H.; Moco, S.I.A.; Lommen, A.; Keurentjes, J.J.B.; Bino, R.J.; Hall, R.D.

    2007-01-01

    Untargeted metabolomics aims to gather information on as many metabolites as possible in biological systems by taking into account all information present in the data sets. Here we describe a detailed protocol for large-scale untargeted metabolomics of plant tissues, based on reversed phase liquid

  7. Introduction to modern liquid chromatography

    National Research Council Canada - National Science Library

    Snyder, Lloyd R; Kirkland, J. J; Dolan, John W

    2010-01-01

    "High-performance liquid chromatography (HPLC) is today the leading technique for chemical analysis and related applications, with an ability to separate, analyze, and/or purify virtually any sample...

  8. Large-scale profiling of diterpenoid glycosides from Stevia rebaudiana using ultrahigh performance liquid chromatography/tandem mass spectrometry.

    Science.gov (United States)

    Shafii, Behnaz; Vismeh, Ramin; Beaudry, Randy; Warner, Ryan; Jones, A Daniel

    2012-07-01

    The plant Stevia rebaudiana accumulates a suite of diterpenoid metabolites that are natural sweeteners finding increased use as sugar substitutes. To guide breeding of stevia plants that accumulate substances with desirable flavor in high yield, rapid and accurate methods are needed to profile these substances in plant populations. This report describes an 8-min ultrahigh performance liquid chromatography-tandem mass spectrometry method for separation and quantification of seven stevia glycosides including steviolbioside; stevioside; rebaudiosides A, B, and C; rubusoside; and dulcoside as well as aglycones steviol and isosteviol. This negative mode electrospray ionization/multiple reaction monitoring method yielded low limits of detection stevia glycosides. Stevioside and Reb A, B, and C were quantified in more than 1,100 extracts from stevia leaves as part of a large-scale profiling exercise. Leaf tissue levels in this population spanned about two orders of magnitude for stevioside (2-125 mg/g dry weight), Reb A (2.5-164 mg/g), Reb B (0.5-50 mg/g), and Reb C (1.5-125 mg/g), but levels of individual metabolites exhibited independent variation. The wide spread of metabolite levels highlights the utility and importance of performing targeted metabolic profiling for large plant populations.

  9. Liquid phase chromatography on microchips

    DEFF Research Database (Denmark)

    Kutter, Jörg Peter

    2012-01-01

    explosive development of, in particular, chromatographic separation systems on microchips, has, however, slowed down in recent years. This review takes a closer, critical look at how liquid phase chromatography has been implemented in miniaturized formats over the past several years, what is important...

  10. Towards Liquid Chromatography Time-Scale Peptide Sequencing and Characterization of Post-Translational Modifications in the Negative-Ion Mode Using Electron Detachment Dissociation Tandem Mass Spectrometry

    DEFF Research Database (Denmark)

    Kjeldsen, Frank; Hørning, Ole B; Jensen, Søren S

    2008-01-01

    Electron detachment dissociation (EDD) of peptide poly-anions is gentle towards post-translational modifications (PTMs) and produces predictable and interpretable fragment ion types (a., x ions). However, EDD is considered an inefficient fragmentation technique and has not yet been implemented...... in large-scale peptide characterization strategies. We successfully increased the EDD fragmentation efficiency (up to 9%), and demonstrate for the first time the utility of EDD-MS/MS in liquid chromatography time-scale experiments. Peptides and phosphopeptides were analyzed in both positive- and negative...

  11. The reproducibility of liquid chromatography separation technology and its potential impact on large scale plant metabolomics experiments.

    Science.gov (United States)

    Arens, Nadja; Döll, Stefanie; Mock, Hans-Peter

    2015-06-01

    Unraveling the constituents of biological samples using HPLC is a central core technology in metabolomics experiments. Consistency in retention time across many samples is a critical criterion for judging the quality of a data set, which must be met before further analysis are possible. Here, the performance of two ultra high-performance liquid chromatography (UHPLC) systems has been compared using an established separation protocol optimized for phenylpropanoids, a class of secondary compounds found in plants displaying intermediate polarity. The two systems differed markedly with respect to their reproducibility and pressure stability. The standard deviation of the retention time of representative peaks differs up to 30-folds between the systems. Adjustments made to the gradient profiles succeeded in equalizing their level of performance. However, the modifications made to the separation protocol reduced the quality of the separation, particularly of the more rapidly eluting components, and lengthened the run time. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. High Performance Liquid Chromatography Method for the ...

    African Journals Online (AJOL)

    High Performance Liquid Chromatography Method for the Determination of Anethole in Rat Plasma. ... Journal Home > Vol 13, No 5 (2014) > ... Results: GC determination showed that anethole in the essential oil of star anise exhibited a ...

  13. Liquid chromatography detection unit, system, and method

    Science.gov (United States)

    Derenzo, Stephen E.; Moses, William W.

    2015-10-27

    An embodiment of a liquid chromatography detection unit includes a fluid channel and a radiation detector. The radiation detector is operable to image a distribution of a radiolabeled compound as the distribution travels along the fluid channel. An embodiment of a liquid chromatography system includes an injector, a separation column, and a radiation detector. The injector is operable to inject a sample that includes a radiolabeled compound into a solvent stream. The position sensitive radiation detector is operable to image a distribution of the radiolabeled compound as the distribution travels along a fluid channel. An embodiment of a method of liquid chromatography includes injecting a sample that comprises radiolabeled compounds into a solvent. The radiolabeled compounds are then separated. A position sensitive radiation detector is employed to image distributions of the radiolabeled compounds as the radiolabeled compounds travel along a fluid channel.

  14. Liquid chromatography of organophosphorus acids

    NARCIS (Netherlands)

    Verweij, A.; Kientz, C.E.; Berg, J. van den

    1988-01-01

    The applicability of different liquid chromatographic systems such as ion-exchange, ion-exclusion, reversed phase and ion-pair partition was studied for the analysis of a number of simple structurally related organophosphorus acids which lack a chromophoric group. Preliminary experiments based on

  15. Porphyrins profile by high performance liquid chromatography ...

    African Journals Online (AJOL)

    Most porphyria symptoms are nonspecific and occur intermittently; resulting frequently in missed diagnosis since the disease itself is a rare one. The aim of the study is to establish a new reliable and accurate laboratory method for separation, identification and quantitation of urinary porphyrins by liquid chromatography ...

  16. determined by high perforiviance liquid chromatography

    African Journals Online (AJOL)

    ABSTRACT. The loss oi" L-ascorbic acid (L-AA) in 14 different cooketi local vegetables found in. Nairobi markets was determined by high performance liquid chromatography. The efifect of quantity of water on the loss of L-AA during cooking was studied with cowpea leaves. It, was found that more. L~AA was lost when ...

  17. Gradient High Performance Liquid Chromatography Method ...

    African Journals Online (AJOL)

    Purpose: To develop a gradient high performance liquid chromatography (HPLC) method for the simultaneous determination of phenylephrine (PHE) and ibuprofen (IBU) in solid dosage form. Methods: HPLC determination was carried out on an Agilent XDB C-18 column (4.6 x 150mm, 5 μ particle size) with a gradient ...

  18. Liquid chromatography-Fourier-transform infrared spectrometry.

    NARCIS (Netherlands)

    Gooijer, C.; Brinkman, U.A.T.; Somsen, G.W.

    1999-01-01

    Over the past years the coupling of liquid chromatography (LC) and Fourier-transform infrared spectrometry (FT-IR) has been pursued primarily to achieve specific detection and/or identification of sample constituents. Two approaches can be discerned in the combination of LC and FT-IR. The first and

  19. Liquid chromatography-Fourier-transform infrared spectrometry

    NARCIS (Netherlands)

    Somsen, G.W; Gooijer, C; Brinkman, U.A Th

    1999-01-01

    Over the past years the coupling of liquid chromatography (LC) and Fourier-transform infrared spectrometry (FT-IR) has been pursued primarily to achieve specific detection and/or identification of sample constituents. Two approaches can be discerned in the combination of LC and FT-IR. The first and

  20. High-Resolution Liquid Chromatography Tandem Mass Spectrometry Enables Large Scale Molecular Characterization of Dissolved Organic Matter

    Directory of Open Access Journals (Sweden)

    Daniel Petras

    2017-12-01

    Full Text Available Dissolved organic matter (DOM is arguably one of the most complex exometabolomes on earth, and is comprised of thousands of compounds, that together contribute more than 600 × 1015 g carbon. This reservoir is primarily the product of interactions between the upper ocean's microbial food web, yet abiotic processes that occur over millennia have also modified many of its molecules. The compounds within this reservoir play important roles in determining the rate and extent of element exchange between inorganic reservoirs and the marine biosphere, while also mediating microbe-microbe interactions. As such, there has been a widespread effort to characterize DOM using high-resolution analytical methods including nuclear magnetic resonance spectroscopy (NMR and mass spectrometry (MS. To date, molecular information in DOM has been primarily obtained through calculated molecular formulas from exact mass. This approach has the advantage of being non-targeted, accessing the inherent complexity of DOM. Molecular structures are however still elusive and the most commonly used instruments are costly. More recently, tandem mass spectrometry has been employed to more precisely identify DOM components through comparison to library mass spectra. Here we describe a data acquisition and analysis workflow that expands the repertoire of high-resolution analytical approaches available to access the complexity of DOM molecules that are amenable to electrospray ionization (ESI MS. We couple liquid chromatographic separation with tandem MS (LC-MS/MS and a data analysis pipeline, that integrates peak extraction from extracted ion chromatograms (XIC, molecular formula calculation and molecular networking. This provides more precise structural characterization. Although only around 1% of detectable DOM compounds can be annotated through publicly available spectral libraries, community-wide participation in populating and annotating DOM datasets could rapidly increase the

  1. Development of suitable method for large-scale urinary glucocorticoid analysis by liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Matsumoto, Akiko; Shimanoe, Chisato; Tanaka, Keitaro; Ichiba, Masayoshi; Hara, Megumi

    2017-07-01

    Levels of urinary glucocorticoids and their concentration ratios have been analyzed as potential markers for various pathological statuses. Large-scale studies may possibly accelerate the investigations; however, a suitable method needs to be established. Analytical conditions for measurement of urinary glucocorticoids with LCMS were examined. Electrospray ionization in the positive ion mode was applied for detection of cortisol (precursor>product ion: 363.3>121.0), cortisol-d4 (internal standard, IS, 367.4>121.1), and cortisone (361.2>163.2). To maximize ionization, acetic acid-ammonium acetate buffer (18mM) at pH 5.3 was employed as eluent A. A C18 column (100mm×2.1mm, 2.7μm) at 50°C was used for the 9.5min binary gradient separation starting with 60% eluent A with methanol being eluent B. Linear correlations were observed between the concentrations and the peak areas in the concentration range of 1-300ng/mL with correlation coefficients (r) of 0.998 and 0.997 for cortisol and cortisone, respectively, without IS adjustment, and 0.999 with IS adjustment for both cortisol and cortisone. Solid-phase extraction (SPE) using a 2mL centrifuge column was performed for the urine samples, with the original and final volumes being 100μL. The SPE of 12 urine specimens could be performed within 30min. The effect of the sample matrix on the quantification of endogenous compounds present in the urine extract was limited (coefficient of variation (CV) of IS-adjusted matrix factor: 4.4-8.1%; urine extracts of 8 individuals); however, substantial peak reduction of cortisol was observed at low concentrations. Exogenous contaminants originating from the SPE centrifuge column seemed to be a main cause for this phenomenon because the pure-water extract showed similar peak reduction. A recovery of ∼50% was obtained for both cortisol and cortisone. Adjustment with the IS improved the apparent recovery, with ∼100% being obtained for both cortisol and cortisone. The recovery rate

  2. Recent development of ionic liquid stationary phases for liquid chromatography.

    Science.gov (United States)

    Shi, Xianzhe; Qiao, Lizhen; Xu, Guowang

    2015-11-13

    Based on their particular physicochemical characteristics, ionic liquids have been widely applied in many fields of analytical chemistry. Many types of ionic liquids were immobilized on a support like silica or monolith as stationary phases for liquid chromatography. Moreover, different approaches were developed to bond covalently ionic liquids onto the supporting materials. The obtained ionic liquid stationary phases show multi-mode mechanism including hydrophobic, hydrophilic, hydrogen bond, anion exchange, π-π, and dipole-dipole interactions. Therefore, they could be used in different chromatographic modes including ion-exchange, RPLC, NPLC and HILIC to separate various classes of compounds. This review mainly summarizes the immobilized patterns and types of ionic liquid stationary phases, their retention mechanisms and applications in the recent five years. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Flow Rates in Liquid Chromatography, Gas Chromatography and Supercritical Fluid Chromatography: A Tool for Optimization

    Directory of Open Access Journals (Sweden)

    Joris Meurs

    2016-08-01

    Full Text Available This paper aimed to develop a standalone application for optimizing flow rates in liquid chromatography (LC, gas chromatography (GC and supercritical fluid chromatography (SFC. To do so, Van Deemter’s equation, Knox’ equation and Golay’s equation were implemented in a MATLAB script and subsequently a graphical user interface (GUI was created. The application will show the optimal flow rate or linear velocity and the corresponding plate height for the set input parameters. Furthermore, a plot will be shown in which the plate height is plotted against the linear flow velocity. Hence, this application will give optimized flow rates for any set conditions with minimal effort.

  4. Recent applications of hydrophilic interaction liquid chromatography in pharmaceutical analysis.

    Science.gov (United States)

    Zhang, Qian; Yang, Feng-Qing; Ge, Liya; Hu, Yuan-Jia; Xia, Zhi-Ning

    2017-01-01

    Hydrophilic interaction liquid chromatography, an alternative liquid chromatography mode, is of particular interest in separating hydrophilic and polar ionic compounds. Compared with traditional liquid chromatography techniques, hydrophilic interaction liquid chromatography offers specific advantages mainly including: (1) relatively green and water-soluble mobile phase composition, which enhances the solubility of hydrophilic and polar ionic compounds; (2) no need for ion-pairing reagents and high content of organic solvent, which benefits mass spectrometry detection; (3) high orthogonality to reverse-phase liquid chromatography, well adapted to two-dimensional liquid chromatography for complicated samples. Therefore, hydrophilic interaction liquid chromatography has been rapidly developed in many areas over the past decades. This review summarizes the recent progress (from 2012 to July 2016) of hydrophilic interaction liquid chromatography in pharmaceutical analysis, with the focus on detecting chemical drugs in various matrices, charactering active compounds of natural products and assessing biotherapeutics through typical structure unit. Moreover, the retention mechanism and behavior of analytes in hydrophilic interaction liquid chromatography as well as some novel hydrophilic interaction liquid chromatography columns used for pharmaceutical analysis are also described. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Liquid chromatography/high resolution tandem mass spectrometry - Tool for the study of polyphenol profile changes during micro-scale biogas digestion of grape marcs.

    Science.gov (United States)

    Kučera, Lukáš; Kurka, Ondřej; Barták, Petr; Bednář, Petr

    2017-01-01

    A microscale discontinuous fermenter was used for anaerobic digestion of wine waste - a hardly gasifiable feedstock material. Efficiency of biogas production, i.e. changes in content of nitrogen, oxygen, carbon dioxide and methane in gas phase, was monitored by gas chromatography/mass spectrometry. Liquid chromatography/high resolution tandem mass spectrometry in combination with principal component analysis and orthogonal projection to latent structures was used to reveal main chemical differences of gasified wine waste mixture from commonly used ones in agricultural biogas plants. Compounds with particular polyphenolic structures appeared among the most distinctive markers. Analysis of samples collected during acidogenic phase and unstabilized methanogenesis indicates formation of certain dihydro-flavonoids in early stages of the process and their consequent degradation. Due to formerly described higher toxicity of some dihydroflavonoids (e.g. taxifolin) compared to their more common counterparts (e.g. quercetin, malvidin etc.), unstabilized digestate would represent a potential environmental risk when used as a fertilizer deserving a proper control. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Optimizing separations in online comprehensive two-dimensional liquid chromatography.

    Science.gov (United States)

    Pirok, Bob W J; Gargano, Andrea F G; Schoenmakers, Peter J

    2017-10-13

    Online comprehensive two-dimensional liquid chromatography has become an attractive option for the analysis of complex nonvolatile samples found in various fields (e.g. environmental studies, food, life, and polymer sciences). Two-dimensional liquid chromatography complements the highly popular hyphenated systems that combine liquid chromatography with mass spectrometry. Two-dimensional liquid chromatography is also applied to the analysis of samples that are not compatible with mass spectrometry (e.g. high-molecular-weight polymers), providing important information on the distribution of the sample components along chemical dimensions (molecular weight, charge, lipophilicity, stereochemistry, etc.). Also, in comparison with conventional one-dimensional liquid chromatography, two-dimensional liquid chromatography provides a greater separation power (peak capacity). Because of the additional selectivity and higher peak capacity, the combination of two-dimensional liquid chromatography with mass spectrometry allows for simpler mixtures of compounds to be introduced in the ion source at any given time, improving quantitative analysis by reducing matrix effects. In this review, we summarize the rationale and principles of two-dimensional liquid chromatography experiments, describe advantages and disadvantages of combining different selectivities and discuss strategies to improve the quality of two-dimensional liquid chromatography separations. © 2017 The Authors. Journal of Separation Science published by WILEY-VCH Verlag GmbH & Co. KGaA.

  7. Comprehensive two-dimensional liquid chromatography: Ion chromatography × reversed-phase liquid chromatography for separation of low-molar-mass organic acids

    NARCIS (Netherlands)

    Brudin, S.S.; Shellie, R.A.; Haddad, P.R.; Schoenmakers, P.J.

    2010-01-01

    In the work presented here a novel approach to comprehensive two-dimensional liquid chromatography is evaluated. Ion chromatography is chosen for the first-dimension separation and reversed-phase liquid chromatography is chosen for the second-dimension separation mode. The coupling of these modes is

  8. Trends in High Performance Liquid Chromatography for Cultural Heritage.

    Science.gov (United States)

    Degano, Ilaria; La Nasa, Jacopo

    2016-04-01

    The separation, detection and quantitation of specific species contained in a sample in the field of Cultural Heritage requires selective, sensitive and reliable methods. Procedures based on liquid chromatography fulfil these requirements and offer a wide range of applicability in terms of analyte types and concentration range. The main applications of High Performance Liquid Chromatography in this field are related to the separation and detection of dyestuffs in archaeological materials and paint samples by reversed-phase liquid chromatography with suitable detectors. The relevant literature will be revised, with particular attention to sample treatment strategies and future developments. Reversed phase chromatography has also recently gained increasing importance in the analysis of lipid binders and lipid materials in archaeological residues: the main advantages and disadvantages of the new approaches will be discussed. Finally, the main applications of ion chromatography and size exclusion chromatography in the field of Cultural Heritage will be revised in this chapter.

  9. Alcoholic fermentation process control by high-performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Morawski, J.; Dincer, A.K.; Ivie, K.

    1983-02-01

    In large-scale fermentation for energy production high-performance liquid chromatography (HPLC) provides an accurate method of monitoring the original oligosaccharides and polysaccharides, as well as their hydrolysis to fermentable monosaccharides. Also measuring the saccharide and alcohol content of the fermentation vat allows overseeing of the process, providing the capability of allowing the fermentation to proceed to the most economical level prior to distillation. Another application for HPLC in a large-scale fermentation for energy is to analyze the stillage for its ethanol content during distillation, in order to observe the efficiency of the still. HPLC can separate and detect very low levels, (i.e., 100 parts per million), of ethanol to yield information concerning the distillation process. These capabilities indicate that HPLC is an extremely useful efficient instrument to the fermentation industries. (Refs. 2).

  10. Instrumentation for hand-portable liquid chromatography.

    Science.gov (United States)

    Sharma, Sonika; Plistil, Alex; Simpson, Robert S; Liu, Kun; Farnsworth, Paul B; Stearns, Stanley D; Lee, Milton L

    2014-01-31

    Liquid chromatography (LC) has lagged behind gas chromatography (GC) in developments related to hand-portable instrumentation. In this work, a new battery-operated (24V DC) nano-flow pumping system with a stop-flow injector was developed and integrated with an on-column UV-absorption detector (254nm) that was reduced in size to an acceptable weight and power usage for field operation. The pumping system, which includes nano-flow pump, stepper motor and high-pressure valve weighs only 1.372kg (3lbs) and can generate up to 110.32MPa (16,000psi) pressure. A major advantage of this pump is that it does not employ a splitter, since it was specifically designed for capillary column use. The volume capacity of the pump is 24μL, and a sample volume as low as 10nL can be injected. Flow rate calibration (300nL to 6.12μL per min) was performed, and an accuracy >99.94% was obtained. The percent injection carry-over was found to be low (RSD 0.31%), which makes it practical for quantitative analysis. The detector linear range and limit of detection (LOD) were determined using sodium anthraquinone-2-sulfonate. A linear regression coefficient (R) of 0.9996 was obtained for a plot of log peak area versus log concentration over the range of 3.2μM to 6.5mM, and the LOD (S/N=3) was found to be 7.8fmol (0.13μM). The short term noise of the detector is comparable to commercially available detectors (∼10(-5)AU). In this work, the system was tested in the laboratory using regular line power (120V AC) with an AC to DC adapter. Reversed-phase isocratic separations were performed using a 15.5cm×75μm i.d. fused silica capillary column containing a monolithic stationary phase synthesized from 1,6-hexanediol dimethacrylate. Good retention time repeatability (RSD 0.09-0.74%) was obtained for a mixture containing an unretained marker (i.e., uracil) and a homologous series of alkyl benzenes. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. ANALYSIS OF AMINO ACIDS BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    Qurat E Noor Baig

    2016-06-01

    Full Text Available Amino acids are the building blocks of proteins and are considered as the key precursors for the formation of hormones and low molecular weight nitrogenous substances with biological importance. Since the analysis of amino acids has been carried out for both qualitative and quantitative purposes with an aim to study their levels in the plasma concentration, the quantitative determination, in particular, also helps in the diagnosis of different diseases associated with their deficiency. This review article deals with the determination of amino acids by chromatographic methods which include ion-exchange chromatography (IEC, high performance liquid chromatography (HPLC, reverse phase-high performance liquid chromatography (RP-HPLC and ultra-performance liquid chromatography (UPLC. The review will also give an idea for the preparation of samples, derivatization methods for the analysis of amino acids (direct and indirect methods and separation of amino acids by high performance liquid chromatographic technique.

  12. Ion Exchange and Liquid Column Chromatography.

    Science.gov (United States)

    Walton, Harold F.

    1980-01-01

    Emphasizes recent advances in principles and methodology in ion exchange and chromatography. Two tables list representative examples for inorganic ions and organic compounds. Cites 544 references. (CS)

  13. Enantioselective high performance liquid chromatography and supercritical fluid chromatography separation of spirocyclic terpenoid flavor compounds.

    Science.gov (United States)

    Schaffrath, Mathias; Weidmann, Verena; Maison, Wolfgang

    2014-10-10

    Chiral spirocyclic terpenoids are abundant natural flavors with significant impact particularly on the food industry. Chromatographic methods for analytical and preparative separation of these compounds are therefore of high interest to natural product chemists in academia and industry. Gas chromatography on chiral stationary phases is currently the standard method for the separation of volatile terpenoids, limiting the scale to analytical quantities. We report herein high performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) protocols for the chiral separation of several racemic spirocyclic terpenoids such as the important flavors theaspirane and vitispirane. A screening of mobile phases and 16 commercially available chiral stationary phases (CSPs) largely based on polysaccharides led to identification of protocols for the separation of all terpenoids tested. SFC methods were found to be particularly useful for the separation of these spirocyclic flavors due to the volatility and low polarity of the compounds. The reported chiral HPLC and SFC protocols are scalable alternatives to gas chromatographic separations of volatile terpenoid flavors. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. High Performance Liquid Chromatography Method for the ...

    African Journals Online (AJOL)

    chromatography (HPLC) technique with UV-VIS detection method was developed for the determination of the compound in rat ... Keywords: Anethole, High performance liguid chromatography, Star anise, Essential oil, Rat plasma,. Illicium verum Hook. .... solution of anethole. Plasma proteins were precipitated by adding 0.3.

  15. Isolation and purification of six iridoid glycosides from gardenia jasminoides fruit by medium-pressure liquid chromatography combined with macroporous resin chromatography.

    Science.gov (United States)

    Wang, Yun; Liu, Hui; Shen, Lifeng; Yao, Lan; Ma, Yinlian; Yu, Dingrong; Chen, Jianhong; Li, Puling; Chen, Ying; Zhang, Cun

    2015-12-01

    Gardeniae fructus is one of the most frequently used herbs in traditional Chinese medicine. In the present study, a process for the enrichment of six iridoid glycosides from Gardeniae fructus was developed using medium-pressure liquid chromatography combined with macroporous resin and reversed-phase chromatography. The purities of different fractions from Gardeniae fructus were assessed using quantitative high-performance liquid chromatography. After fractionation using HPD-100 column chromatography, a 30% ethanol fraction was selected based on high-performance liquid chromatography and liquid chromatography with mass spectrometry qualitative analysis to separate and purify. Based on the orientation analysis results, six compounds-deacetyl asperulosidic acid methyl ester, gardenoside, ixoroside, scandoside methyl ester, genipin-1-O-β-d-gentiobioside, and geniposide-were successfully isolated and purified in three to four combined steps from Gardeniae fructus. The purities of these compounds were found by high-performance liquid chromatography analysis to be 97.9, 98.1, 95.5, 96.3, 97.1, and 98.7%, respectively. Moreover, their structures were elucidated by NMR spectroscopy and liquid chromatography with tandem mass spectrometry. The separation process was highly efficient, rapid, and accurate, making it a potential approach for the large-scale production of iridoids in the laboratory and providing several marker compounds for quality control. This procedure may be meaningful for the purification of other natural products used in traditional Chinese medicine. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Improved sensitivity using liquid chromatography mass spectrometry ...

    African Journals Online (AJOL)

    Triple quadrupole mass spectrometry (MS/MS) was used to confirm the identity of BMAA in cyanobacteria based on product ions. We show a 10-fold increase in sensitivity with the LC-MS method compared to the previously published gas chromatography mass spectrometry (GC-MS) method with pre-column derivatised ...

  17. Analysis of Tocopherols by High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    B. Edison

    2009-01-01

    Full Text Available : Gas chromatography is the key technique for organic components and also for tocopherols analysis. High performance liquid chromatography has an important role to take part in applications such as the handling of less usual samples, prevention of degradation of heat sensitive functional groups and for micro preparative purposes. Many approaches for development of improved methods are suggested, especially for reversed phase applications.

  18. Liquid Chromatography Applied to Space System

    Science.gov (United States)

    Poinot, Pauline; Chazalnoel, Pascale; Geffroy, Claude; Sternberg, Robert; Carbonnier, Benjamin

    Searching for signs of past or present life in our Solar System is a real challenge that stirs up the curiosity of scientists. Until now, in situ instrumentation was designed to detect and determine concentrations of a wide number of organic biomarkers. The relevant method which was and still is employed in missions dedicated to the quest of life (from Viking to ExoMars) corresponds to the pyrolysis-GC-MS. Along the missions, this approach has been significantly improved in terms of extraction efficiency and detection with the use of chemical derivative agents (e.g. MTBSTFA, DMF-DMA, TMAH…), and in terms of analysis sensitivity and resolution with the development of in situ high-resolution mass spectrometer (e.g. TOF-MS). Thanks to such an approach, organic compounds such as amino acids, sugars, tholins or polycyclic aromatic hydrocarbons (PAHs) were expected to be found. However, while there’s a consensus that the GC-MS of Viking, Huygens, MSL and MOMA space missions worked the way they had been designed to, pyrolysis is much more in debate (Glavin et al. 2001; Navarro-González et al. 2006). Indeed, (1) it is thought to remove low levels of organics, (2) water and CO2 could interfere with the detection of likely organic pyrolysis products, and (3) only low to mid-molecular weight organic molecules can be detected by this technique. As a result, researchers are now focusing on other in situ techniques which are no longer based on the volatility of the organic matter, but on the liquid phase extraction and analysis. In this line, micro-fluidic systems involving sandwich and/or competitive immunoassays (e.g. LMC, SOLID; Parro et al. 2005; Sims et al. 2012), micro-chip capillary electrophoreses (e.g. MOA; Bada et al. 2008), or nanopore-based analysis (e.g. BOLD; Schulze-Makuch et al. 2012) have been conceived for in situ analysis. Thanks to such approaches, molecular biological polymers (polysaccharides, polypeptides, polynucleotides, phospholipids, glycolipids

  19. Liquid chromatography and liquid chromatography-mass spectrometry analysis of donepezil degradation products

    Directory of Open Access Journals (Sweden)

    Mladenović Aleksandar R.

    2015-01-01

    Full Text Available This study describes the investigation of degradation products of donepezil (DP using stability indicating RP-HPLC method for determination of donepezil, which is a centrally acting reversible acetylcholinesterase inhibitor. In order to investigate the stability of drug and formed degradation products, a forced degradation study of drug sample and finished product under different forced degradation conditions has been conducted. Donepezil hydrochloride and donepezil tablets were subjected to stress degradation conditions recommended by International Conference on Harmonization (ICH. Donepezil hydrochloride solutions were subjected to acid and alkali hydrolysis, chemical oxidation and thermal degradation. Significant degradation was observed under alkali hydrolysis and oxidative degradation conditions. Additional degradation products were observed under the conditions of oxidative degradation. The degradation products observed during forced degradation studies were monitored using the high performance liquid chromatography (HPLC method developed. The parent method was modified in order to obtain LC-MS compatible method which was used to identify the degradation products from forced degradation samples using high resolution mass spectrometry. The mass spectrum provided the precise mass from which derived molecular formula of drug substance and degradation products formed and proved the specificity of the method unambiguously. [Projekat Ministarstva nauke Republike Srbije, br. 172013

  20. Instrumental liquid chromatography: a practical manual on high-performance liquid chromatographic methods

    National Research Council Canada - National Science Library

    Parris, N. A

    1976-01-01

    Available texts on liquid chromatography have tended to emphasize the developments in the theoretical understanding of the technique and methodology or to list numerous applications, complete with experimental details...

  1. Challenges in polymer analysis by liquid chromatography

    NARCIS (Netherlands)

    Uliyanchenko, E.; van der Wal, S.; Schoenmakers, P.J.

    2012-01-01

    Synthetic polymers are very important in our daily life. Many valuable properties of polymers are determined by their molecular weight and chemical composition. Liquid chromatographic (LC) techniques are very commonly used for molecular characterisation of polymers. LC analysis of macromolecules is

  2. Liquid Chromatography – Mass Spectrometry Method for the ...

    African Journals Online (AJOL)

    Purpose: To develop and validate a simple and selective high performance liquid chromatography photo diode array mass spectrometry (HPLC-PDA-MS/MS) method for simultaneous determination and confirmation of seven major active alkaloids (6-Hydroxy-ß-Carboline-1-carboxylic acid, ß-Carboline-1- carboxylic acid, ...

  3. The high performance liquid chromatography (HPLC) analysis of ...

    African Journals Online (AJOL)

    The high performance liquid chromatography (HPLC) analysis of ultraviolet (UV) irradiated chlorophyll a and secondary plant compounds. ... in all the samples leaving a bleached extract suitable for biological assays. Key words: Chlorophyll a, UV radiation, activated charcoal, HPLC, secondary compounds in plant extracts.

  4. Quantification of Tea Flavonoids by High Performance Liquid Chromatography

    Science.gov (United States)

    Freeman, Jessica D.; Niemeyer, Emily D.

    2008-01-01

    We have developed a laboratory experiment that uses high performance liquid chromatography (HPLC) to quantify flavonoid levels in a variety of commercial teas. Specifically, this experiment analyzes a group of flavonoids known as catechins, plant-derived polyphenolic compounds commonly found in many foods and beverages, including green and black…

  5. Characterization of porous silicon integrated in liquid chromatography chips

    NARCIS (Netherlands)

    Tiggelaar, Roald M.; Verdoold, Vincent; Eghbali, H.; Desmet, G.; Gardeniers, Johannes G.E.

    2009-01-01

    Properties of porous silicon which are relevant for use of the material as a stationary phase in liquid chromatography chips, like porosity, pore size and specific surface area, were determined with high-resolution SEM and N2 adsorption–desorption isotherms. For the anodization conditions

  6. An optimized gossypol high-performance liquid chromatography ...

    Indian Academy of Sciences (India)

    A comparative study on gossypol content of various genetic types of pigment glands of cotton varieties was conducted through an optimized high-performance liquid chromatography (HPLC) on a C18 column (4.6 mm × 250 mm, 5 m particle) with methanol–0.5% acetic acid aqueous solution, 90 : 10 (v/v), as mobile phase, ...

  7. Coupling of column liquid chromatography and Fourier transform infrared spectrometry.

    NARCIS (Netherlands)

    Gooijer, C.; Velthorst, N.H.; Brinkman, U.A.T.; Somsen, G.W.

    1998-01-01

    This paper provides an extensive overview of the literature on the coupling of column liquid chromatography (LC) and Fourier transform infrared spectrometry (FT-IR). Flow-cell-based FT-IR detection and early solvent- elimination interfaces for LC-FT-IR are discussed in brief. A comprehensive

  8. Coupling of column liquid chromatography and Fourier transform infrared spectrometry

    NARCIS (Netherlands)

    Somsen, G.W; Gooijer, C; Velthorst, N.H; Brinkman, U.A Th

    1998-01-01

    This paper provides an extensive overview of the literature on the coupling of column liquid chromatography (LC) and Fourier transform infrared spectrometry (FT-IR). Flow-cell-based FT-IR detection and early solvent-elimination interfaces for LC-FT-IR are discussed in brief. A comprehensive

  9. Alternative solvents can make preparative liquid chromatography greener

    NARCIS (Netherlands)

    Shen, Y.; Chen, B.; Beek, van T.A.

    2015-01-01

    To make preparative Reversed-Phase High Performance Liquid Chromatography (RP-pHPLC) greener, alternative solvents were considered among others in terms of toxicity, cost, safety, workability, chromatographic selectivity and elution strength. The less toxic solvents ethanol, acetone and ethyl

  10. New Highly-Sensitive Ultra-Performance Liquid Chromatography ...

    African Journals Online (AJOL)

    Purpose: To develop and validate a simple, rapid, sensitive and specific ultraperformance liquid chromatography mass spectrometry method for the quantification of the angiotensin II receptor antagonist, telmisartan (TEL), in human plasma. Methods: After simple protein precipitation using acetonitrile and methanol, TEL and ...

  11. Towards early detection of the hydrolytic degradation of poly(bisphenol A)carbonate by hyphenated liquid chromatography and comprehensive two-dimensional liquid chromatography

    NARCIS (Netherlands)

    Coulier, L.; Kaal, E.R.; Hankemeier, Th.

    2006-01-01

    The hydrolytic degradation of poly(bisphenol A)carbonate (PC) has been characterized by various liquid chromatography techniques. Size exclusion chromatography (SEC) showed a significant decrease in molecular mass as a result of hydrolytic degradation, while 'liquid chromatography at critical

  12. Detection of Griseofulvin and Dechlorogriseofulvin by Thin-Layer Chromatography and Gas-Liquid Chromatography

    Science.gov (United States)

    Cole, R. J.; Kirksey, J. W.; Holaday, C. E.

    1970-01-01

    A rapid and accurate method is described for the determination of griseofulvin and dechlorogriseofulvin extracted from Penicillium urticae with chloroform. Thinlayer chromatography was used to tentatively identify griseofulvin or dechlorogriseofulvin, or both. Two gas-liquid chromatographic systems provided additional qualitative information and simultaneous quantitation of the individual compounds. PMID:5415206

  13. On-line coupling of column liquid chromatography and Raman spectroscopy using a liquid core waveguide,

    NARCIS (Netherlands)

    Dijkstra, R.J.; Bader, A.N.; Brinkman, U.A.T.; Gooijer, C.; Hoornweg, G.Ph.

    1999-01-01

    The on-line coupling of Raman spectroscopy and conventional-size liquid chromatography (LC) by using a plastic liquid core waveguide (CW) was explored. The LCW considered (length 30 cm, internal diameter 280 μm, internal volume 18 μL) is suitable for light guiding through aqueous LC eluents since

  14. Optimizing heterosurface adsorbent synthesis for liquid chromatography

    Science.gov (United States)

    Bogoslovskii, S. Yu.; Serdan, A. A.

    2016-03-01

    The structural and geometric parameters of a silica matrix (SM) for the synthesis of heterosurface adsorbents (HAs) are optimized. Modification is performed by shielding the external surfaces of alkyl-modified silica (AS) using human serum albumin and its subsequent crosslinking. The structural and geometric characteristics of the SM, AS, and HA are measured via low-temperature nitrogen adsorption. It is found that the structural characteristics of AS pores with diameters D 9 nm reduces significantly due to adsorption of albumin. It is concluded that silica gel with a maximum pore size distribution close to 5 nm and a minimal proportion of pores with D > 9 nm is optimal for HA synthesis; this allows us to achieve the greatest similarity between the chromatographic retention parameters for HA and AS. The suitability of the synthesized adsorbents for analyzing drugs in biological fluids through direct sample injection is confirmed by chromatography. It was found that the percentage of the protein fraction detected at the outlet of the chromatographic column is 98%.

  15. Theories to support method development in comprehensive two-dimensional liquid chromatography - A review

    NARCIS (Netherlands)

    Bedani, F.; Schoenmakers, P.J.; Janssen, H.-G.

    2012-01-01

    On-line comprehensive two-dimensional liquid chromatography techniques promise to resolve samples that current one-dimensional liquid chromatography methods cannot adequately deal with. To make full use of the potential of two-dimensional liquid chromatography, optimization is required. Optimization

  16. 21 CFR 862.2250 - Gas liquid chromatography system for clinical use.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Gas liquid chromatography system for clinical use... Instruments § 862.2250 Gas liquid chromatography system for clinical use. (a) Identification. A gas liquid chromatography system for clinical use is a device intended to separate one or more drugs or compounds from a...

  17. 21 CFR 862.2260 - High pressure liquid chromatography system for clinical use.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false High pressure liquid chromatography system for... Clinical Laboratory Instruments § 862.2260 High pressure liquid chromatography system for clinical use. (a) Identification. A high pressure liquid chromatography system for clinical use is a device intended to separate...

  18. Chromatography

    Science.gov (United States)

    ... that are bonded together. For example, water is a chemical bond of oxygen and hydrogen. Proteins are another type of chemical compound. There are different kinds of chromatography. These include gas, high pressure liquid, or ion ...

  19. Standard flow liquid chromatography for shotgun proteomics in bioenergy research

    Directory of Open Access Journals (Sweden)

    Susana M. González Fernández-Niño

    2015-04-01

    Full Text Available Over the past ten years the bioenergy and biofuels field has realized significant achievements that have encouraged many follow on efforts centered on biosynthetic production of fuel-like compounds. Key to the success of these efforts has been transformational developments in feedstock characterization and metabolic engineering of biofuel-producing microbes. Lagging far behind these advancements are analytical methods to characterize and quantify systems of interest to the bioenergy field. In particular the utilization of proteomics, while valuable for identifying novel enzymes and diagnosing problems associated with biofuel-producing microbes, is limited by a lack of robustness and limited throughput. Nano-flow liquid chromatography coupled to high-mass accuracy, high-resolution mass spectrometers has become the dominant approach for the analysis of complex proteomic samples, yet such assays still require dedicated experts for data acquisition, analysis, and instrument upkeep. The recent adoption of standard flow chromatography (ca. 0.5 mL/min for targeted proteomics has highlighted the robust nature and increased throughput of this approach for sample analysis. Consequently, we assessed the applicability of standard flow liquid chromatography for shotgun proteomics using samples from Escherichia coli and Arabidopsis thaliana, organisms commonly used as model systems for lignocellulosic biofuels research. Employing 120 minute gradients with standard flow chromatography we were able to routinely identify nearly 800 proteins from E. coli samples, while for samples from Arabidopsis over 1,000 proteins could be reliably identified. An examination of identified peptides indicated that the method was suitable for reproducible applications in shotgun proteomics. Standard flow liquid chromatography for shotgun proteomics provides a robust approach for the analysis of complex samples. To the best of our knowledge this study represents the first attempt

  20. An electrostatic precipitator for preparative gas-liquid chromatography.

    Science.gov (United States)

    Borka, L; Privett, O S

    1966-03-01

    The effect of the operating variables of electrostatic precipitators on the recovery and structure of methyl esters and related aerosol forming compounds collected in preparative gas-liquid chromatography was studied.Aerosol formation was prevented by AC or DC voltages of 5000 to 12000 volts. AC was more effective than DC but caused changes in structure which were detectable by both thin-layer and gas-liquid chromatographic methods of analysis.An apparatus of simple construction and operation was designed for the collection of methyl esters and its use demonstrated with several model compounds.

  1. Protein separations using enhanced-fluidity liquid chromatography.

    Science.gov (United States)

    Bennett, Raffeal; Olesik, Susan V

    2017-11-10

    Enhanced-fluidity liquid chromatography (EFLC) methods using methanol/H 2 O/CO 2 and hydrophilic interaction liquid chromatography (HILIC) were explored for the separation of proteins and peptides. EFLC is a separation mode that uses a mobile phase made of conventional solvents combined with liquid carbon dioxide (CO 2 ) in subcritical conditions. The addition of liquid CO 2 enhances diffusivity and decreases viscosity while maintaining mixture polarity, which typically results in reduced time of analysis. TFA additive and elevated temperature were leveraged as key factors in the separation of a 13-analyte intact protein mixture in under 5min. Under these conditions EFLC showed modest improvement in terms of peak asymmetry and analysis time over the competing ACN/H 2 O separation. Protein analytes detected by electrospray ionization - quadrupole time of flight, were shown to be unaffected by the addition of CO 2 in the mobile phase. Herein, the feasibility of separating hydrophilic proteins up to 80kDa (with transferrin) is demonstrated for CO 2 -containing mobile phases. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Comprehensive two-dimensional liquid chromatography and hyphenated liquid chromatography to study the degradation of poly(bisphenol A)carbonate

    NARCIS (Netherlands)

    Coulier, L.; Kaal, E.R.; Hankemeier, T.

    2005-01-01

    Size exclusion chromatography (SEC), gradient polymer elution chromatography (GPEC) and liquid chromatography at critical conditions (LC-CC) have been developed and applied to observe chemical changes in poly(bisphenol A)carbonate (PC) due to hydrolytic degradation. Especially LC-CC appeared to be

  3. Size distributions of gold nanoclusters studied by liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    WILCOXON,JESS P.; MARTIN,JAMES E.; PROVENCIO,PAULA P.

    2000-05-23

    The authors report high pressure liquid chromatography, (HPLC), and transmission electron microscopy, (TEM), studies of the size distributions of nanosize gold clusters dispersed in organic solvents. These metal clusters are synthesized in inverse micelles at room temperature and those investigated range in diameter from 1--10 nm. HPLC is sensitive enough to discern changes in hydrodynamic volume corresponding to only 2 carbon atoms of the passivating agent or metal core size changes of less than 4 {angstrom}. The authors have determined for the first time how the total cluster volume (metal core + passivating organic shell) changes with the size of the passivating agent.

  4. On the Density Scaling of Liquid Dynamics

    Science.gov (United States)

    2011-01-01

    squalane in Fig. 1, which is representative of the literature results for dielectric relaxation times of supercooled liquids and polymers. Many liquids...reduced viscosity (filled symbols) of squalane . The data extend over many decades of viscosity, therefore both quantities scale with identical scaling

  5. Reversed-phase liquid chromatography and argentation chromatography of the minor capsaicinoids.

    Science.gov (United States)

    Thompson, Robert Q; Phinney, Karen W; Sander, Lane C; Welch, Michael J

    2005-04-01

    An investigation of the liquid chromatography of the minor capsaicinoids in a commercial capsaicinoid mixture is reported. Twelve stationary phases including C8, C18, C30, phenyl, and cation-exchange chemistries were examined in combination with isocratic aqueous methanol and aqueous acetonitrile mobile phases. A phenyl stationary phase and aqueous acetonitrile mobile phase baseline-resolved 7 of 11 capsaicinoids, and selected ion chromatograms (LC-ESI-MS) demonstrated this was the most effective reversed-phase separation. Argentation chromatography with an alkyl or phenyl column and aqueous silver nitrate-methanol mobile phase revealed the presence of the 6-ene-8-methyl and 6-ene-9-methyl homocapsaicin isomers and the absence of 7-ene-9-methyl homocapsaicin. A mixed phenyl-cation-exchange stationary phase (charged with silver ion) enabled unique and useful separations of the capsaicinoids.

  6. Comparison of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography for the separation of synthetic cathinones.

    Science.gov (United States)

    Carnes, Stephanie; O'Brien, Stacey; Szewczak, Angelica; Tremeau-Cayel, Lauriane; Rowe, Walter F; McCord, Bruce; Lurie, Ira S

    2017-09-01

    A comparison of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography for the separation of synthetic cathinones has been conducted. Nine different mixtures of bath salts were analyzed in this study. The three different chromatographic techniques were examined using a general set of controlled synthetic cathinones as well as a variety of other synthetic cathinones that exist as positional isomers. Overall 35 different synthetic cathinones were analyzed. A variety of column types and chromatographic modes were examined for developing each separation. For the ultra high performance supercritical fluid chromatography separations, analyses were performed using a series of Torus and Trefoil columns with either ammonium formate or ammonium hydroxide as additives, and methanol, ethanol or isopropanol organic solvents as modifiers. Ultra high performance liquid chromatographic separations were performed in both reversed phase and hydrophilic interaction chromatographic modes using SPP C18 and SPP HILIC columns. Gas chromatography separations were performed using an Elite-5MS capillary column. The orthogonality of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography was examined using principal component analysis. For the best overall separation of synthetic cathinones, the use of ultra high performance supercritical fluid chromatography in combination with gas chromatography is recommended. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. (Vapour + liquid) equilibria of ternary systems with ionic liquids using headspace gas chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Mokhtarani, Babak [Chemistry and Chemical Engineering Research Center of Iran, P.O. Box 14335-186, Tehran (Iran, Islamic Republic of); Gmehling, Juergen, E-mail: gmehling@tech.chem.uni-oldenburg.d [Carl von Ossietzky Universitaet Oldenburg, Technische Chemie, D-26111 Oldenburg (Germany)

    2010-08-15

    (Vapour + liquid) equilibrium (VLE) data for the ternary systems (hexane + benzene), (hexane + cyclohexane), (benzene + cyclohexane), and (ethanol + water) with an ionic liquid as entrainer for extractive distillation were measured by headspace gas chromatography. As ionic liquids, 1-hexyl-3-methyl-imidazolium bis (trifluoromethyl-sulfonyl) imide [HMIM][BTI], 1-octyl-3-methyl-imidazolium bis (trifluoromethyl-sulfonyl) imide [OMIM][BTI], 1-octyl-3-methyl-imidazolium trifluoro-methanesulfonate [OMIM][OTF], and 1-butyl-3-methyl-imidazolium trifluoro-methanesulfonate [BMIM][OTF] were used. The experimental data show that the ionic liquids investigated have a great influence on the separation factors of the systems (hexane + benzene), (hexane + cyclohexane), and (benzene + cyclohexane). The experimental data were compared with the predicted results using mod. UNIFAC (Do). The predicted results are in good agreement with the experimental data.

  8. Suppression of deleterious effects of free silanols in liquid chromatography by imidazolium tetrafluoroborate ionic liquids.

    Science.gov (United States)

    Kaliszan, Roman; Marszałł, Michał Piotr; Markuszewski, Michał Jan; Baczek, Tomasz; Pernak, Juliusz

    2004-03-19

    Silica-based stationary phases are commonly used in liquid chromatography, but their surface acidity causes known problems, especially when separating basic compounds. Deleterious effects of free silanols are not fully removed by standard prevention procedures consisting in adding alkylamines or other amino quenchers to the eluents. We found that ionic liquids of the imidazolium tetrafluoroborate class, added to mobile phases at concentrations of 0.5-1.5% (v/v), blocked silanols and provided excellent thin-layer chromatographic separations of strongly basic drugs which were otherwise not eluted, even with neat acetonitrile as the mobile phase. The silanol suppressing potency of imidazolium tetrafluoroborates was demonstrated to markedly exceed that of the standard mobile phase additives, like triethylamine, dimethyloctylamine and ammonia. The proposed new mobile phase additives were also demonstrated to provide reliable lipophilicity parameters of base drug analytes as determined by gradient mode of high-performance liquid chromatography. By applying the readily available and environmentally friendly imidazolium tetrafluoroborate ionic liquids, simple and efficient means of improvement of liquid chromatographic analysis of organic bases were elaborated.

  9. Ultrafast Chiral Chromatography as the Second Dimension in Two-Dimensional Liquid Chromatography Experiments.

    Science.gov (United States)

    Barhate, Chandan L; Regalado, Erik L; Contrella, Nathan D; Lee, Joon; Jo, Junyong; Makarov, Alexey A; Armstrong, Daniel W; Welch, Christopher J

    2017-03-21

    Chromatographic separation and analysis of complex mixtures of closely related species is one of the most challenging tasks in modern pharmaceutical analysis. In recent years, two-dimensional liquid chromatography (2D-LC) has become a valuable tool for improving peak capacity and selectivity. However, the relatively slow speed of chiral separations has limited the use of chiral stationary phases (CSPs) as the second dimension in 2D-LC, especially in the comprehensive mode. Realizing that the recent revolution in the field of ultrafast enantioselective chromatography could now provide significantly faster separations, we herein report an investigation into the use of ultrafast chiral chromatography as a second dimension for 2D chromatographic separations. In this study, excellent selectivity, peak shape, and repeatability were achieved by combining achiral and chiral narrow-bore columns (2.1 mm × 100 mm and 2.1 mm × 150 mm, sub-2 and 3 μm) in the first dimension with 4.6 mm × 30 mm and 4.6 mm × 50 mm columns packed with highly efficient chiral selectors (sub-2 μm fully porous and 2.7 μm fused-core particles) in the second dimension, together with the use of 0.1% phosphoric acid/acetonitrile eluents in both dimensions. Multiple achiral × chiral and chiral × chiral 2D-LC examples (single and multiple heart-cutting, high-resolution sampling, and comprehensive) using ultrafast chiral chromatography in the second dimension are successfully applied to the separation and analysis of complex mixtures of closely related pharmaceuticals and synthetic intermediates, including chiral and achiral drugs and metabolites, constitutional isomers, stereoisomers, and organohalogenated species.

  10. Determination of sulfonamides in butter samples by ionic liquid magnetic bar liquid-phase microextraction high-performance liquid chromatography.

    Science.gov (United States)

    Wu, Lijie; Song, Ying; Hu, Mingzhu; Xu, Xu; Zhang, Hanqi; Yu, Aimin; Ma, Qiang; Wang, Ziming

    2015-01-01

    A novel, simple, and environmentally friendly pretreatment method, ionic liquid magnetic bar liquid-phase microextraction, was developed for the determination of sulfonamides in butter samples by high-performance liquid chromatography. The ionic liquid magnetic bar was prepared by inserting a stainless steel wire into the hollow of a hollow fiber and immobilizing ionic liquid in the micropores of the hollow fiber. In the extraction process, the ionic liquid magnetic bars were used to stir the mixture of sample and extraction solvent and enrich the sulfonamides in the mixture. After extraction, the analyte-adsorbed ionic liquid magnetic bars were readily isolated with a magnet from the extraction system. It is notable that the present method was environmentally friendly since water and only several microliters of ionic liquid were used in the whole extraction process. Several parameters affecting the extraction efficiency were investigated and optimized, including the type of ionic liquid, sample-to-extraction solvent ratio, the number of ionic liquid magnetic bars, extraction temperature, extraction time, salt concentration, stirring speed, pH of the extraction solvent, and desorption conditions. The recoveries were in the range of 73.25-103.85 % and the relative standard deviations were lower than 6.84 %. The experiment results indicated that the present method was effective for the extraction of sulfonamides in high-fat content samples.

  11. Scaling of viscous dynamics in simple liquids

    DEFF Research Database (Denmark)

    Bøhling, Lasse; Ingebrigtsen, Trond; Grzybowski, A.

    2012-01-01

    Supercooled liquids are characterized by relaxation times that increase dramatically by cooling or compression. From a single assumption follows a scaling law according to which the relaxation time is a function of h(ρ) over temperature, where ρ is the density and the function h(ρ) depends...... on the liquid in question. This scaling is demonstrated to work well for simulations of the Kob–Andersen binary Lennard-Jones mixture and two molecular models, as well as for the experimental results for two van der Waals liquids, dibutyl phthalate and decahydroisoquinoline. The often used power-law density...... scaling, h(ρ)∝ργ, is an approximation to the more general form of scaling discussed here. A thermodynamic derivation was previously given for an explicit expression for h(ρ) for liquids of particles interacting via the generalized Lennard-Jones potential. Here a statistical mechanics derivation is given...

  12. A rapid liquid chromatography determination of free formaldehyde in cod.

    Science.gov (United States)

    Storey, Joseph M; Andersen, Wendy C; Heise, Andrea; Turnipseed, Sherri B; Lohne, Jack; Thomas, Terri; Madson, Mark

    2015-01-01

    A rapid method for the determination of free formaldehyde in cod is described. It uses a simple water extraction of formaldehyde which is then derivatised with 2,4-dinitrophenylhydrazine (DNPH) to form a sensitive and specific chromophore for high-performance liquid chromatography (HPLC) detection. Although this formaldehyde derivative has been widely used in past tissue analysis, this paper describes an improved derivatisation procedure. The formation of the DNPH formaldehyde derivative has been shortened to 2 min and a stabilising buffer has been added to the derivative to increase its stability. The average recovery of free formaldehyde in spiked cod was 63% with an RSD of 15% over the range of 25-200 mg kg(-1) (n = 48). The HPLC procedure described here was also compared to a commercial qualitative procedure - a swab test for the determination of free formaldehyde in fish. Several positive samples were compared by both methods.

  13. Separation of human tear proteins by high performance liquid chromatography.

    Science.gov (United States)

    Boonstra, A; Kijlstra, A

    1984-12-01

    The optimal conditions for separating human tear proteins by high performance liquid chromatography (HPLC) using a Waters I-125 gel filtration column were investigated. Several elution buffers were tested including phosphate buffer alone and phosphate buffer to which varying amounts of NaCl or 0.1% Tween was added. The combination of phosphate buffer (pH 5.28), 0.5 M NaCl and 0.1% Tween gave the best resolution and a recovery of 90% of the proteins applied. Tear lactoferrin was shown to adhere to the column packing when the molarity of the elution buffer was not high enough. Using optimal conditions, the tear proteins IgA, lactoferrin and lysozyme were identified in distinct peaks after a preparative HPLC run. When used in combination with Schirmer strips as a tear sampling method, HPLC was shown to be a rapid, simple and reproducible way of investigating the composition of tear proteins.

  14. Biological Fingerprinting of Herbal Samples by Means of Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Łukasz Cieśla

    2012-01-01

    Full Text Available Biological chromatographic fingerprinting is a relatively new concept in the quality control of herbal samples. Originally it has been developed with the application of HPLC, and recently herbal samples' biological profiles have been obtained by means of thin-layer chromatography (TLC. This paper summarizes the application of liquid chromatographic techniques for the purpose of biological fingerprint analysis (BFA of complex herbal samples. In case of biological TLC fingerprint, which is a relatively novel solution, perspectives of its further development are outlined in more detail. Apart from already published data, some novel results are also shown and briefly discussed. The paper aims at drawing scientists' attention to the unique solutions offered by biological fingerprint construction.

  15. [Analysis of benzodiazepine derivative mixture by gas-liquid chromatography].

    Science.gov (United States)

    Zevzikovas, Andrejus; Kiliuviene, Guoda; Ivanauskas, Liudas; Dirse, Vidmantas

    2002-01-01

    The analysis of mixture of benzodiazepine derivates (chlordiazepoxide, flunitrazepam, medazepam, nitrazepam, oxazepam and tetrazepam) by gas--liquid chromatography (GLC) in purpose to separate and identify these psychotropic drugs in mixture is presented in this article. The experiment was carried out in vitro, accommodating this method for identification and separation of drugs, isolated from biological objects (blood and urine). Referring to data of annual reports of chemical investigations (1) above-mentioned psychotropic drugs are very frequent among drug intoxication. In most cases they are detected in the mixture of the same or different pharmacological group, and this causes difficulty for separation and identification. The analysis of the mixture was carried out by GLC, which is widely used in practice of forensic-chemical examination. Adsorbents and stationery phases were changed; the conditions and parameters of chromatography were modified, in purpose totally separate preparations in the mixture. For the separation and identification of all three preparation the column packed with Inerton Super with stationary phase 3% OV-17 is suitable. The column temperature-290 degrees C. The mixture of these drugs was excreted from body fluids (blood and urine) in vitro and investigated by GLC under these conditions. The results of investigation were similar.

  16. Group type analysis of asphalt by column liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, C.; Yang, J.; Xue, Y.; Li, Y. [Chinese Academy of Science, Taiyuan (China)

    2008-07-01

    An improved analysis method for characterization of asphalt was established. The method is based on column chromatography technique. The asphalts were separated into four groups: saturates, aromatics, resins, and asphaltenes, quantitatively. About 0.1 g of sample was required in each analysis. About 20 mL of n-heptanes was used to separate out saturates first. Then about 35 mL of n-heptanes/dichloromethane (.5, v/v) mixture was used to separate out aromatics. About 30 mL of dichloromethane/tetrahydrofuran (1/3, v/v) mixture was used to separate out resin. The quality of the separation was confirmed by infrared spectra (IR) and {sup 1}H NMR analysis. The model compounds, tetracosan for saturates, dibenz(o)anthracen for aromatics, and acetanilide for resins were used for verification. The IR and {sup 1}H NMR analysis of the prepared fractions from the column liquid chromatography were in good agreement that of pure reagents.

  17. Recent advances in ultra-high performance liquid chromatography for the analysis of traditional chinese medicine

    Science.gov (United States)

    Traditional Chinese medicines (TCMs) have been widely used for the prevention and treatment of various diseases for thousands of years in China. Ultra Performance Liquid Chromatography (UHPLC) is a relatively new technique offering new possibilities in liquid chromatography. This paper reviews recen...

  18. Photoionisation detection in packed-capillary liquid and supercritical-fluid chromatography.

    NARCIS (Netherlands)

    Zegers, B.N.; Hessels, R.; Jagesar, J.; Rozenbrand, J.; Lingeman, H.; Brinkman, U.A.T.

    1995-01-01

    A photoionisation detector (PID) was coupled to packed-capillary liquid and supercritical-fluid chromatography to study its performance. Several mobile phases were tested to evaluate the potential of liquid chromatography with photoionisation detection, LC-PID. The behaviour of the PID was not as

  19. Investigating the Retention Mechanisms of Liquid Chromatography Using Solid-Phase Extraction Cartridges

    Science.gov (United States)

    O'Donnell, Mary E.; Musial, Beata A.; Bretz, Stacey Lowery; Danielson, Neil D.; Ca, Diep

    2009-01-01

    Liquid chromatography (LC) experiments for the undergraduate analytical laboratory course often illustrate the application of reversed-phase LC to solve a separation problem, but rarely compare LC retention mechanisms. In addition, a high-performance liquid chromatography instrument may be beyond what some small colleges can purchase. Solid-phase…

  20. Salting-out assisted liquid-liquid extraction coupled to dispersive liquid-liquid microextraction for the determination of chlorophenols in wine by high-performance liquid chromatography.

    Science.gov (United States)

    Fan, Yingying; Hu, Shibin; Liu, Shuhui

    2014-12-01

    A novel procedure of sample preparation combined with high-performance liquid chromatography with diode array detection is introduced for the analysis of highly chlorinated phenols (trichlorophenols, tetrachlorophenols, and pentachlorophenol) in wine. The main features of the proposed method are (i) low-toxicity diethyl carbonate as extraction solvent to selectively extract the analytes without matrix effect, (ii) the combination of salting-out assisted liquid-liquid extraction and dispersive liquid-liquid microextraction to achieve an enrichment factor of 334-361, and (iii) the extract is analyzed by high-performance liquid chromatography to avoid derivatization. Under the optimum conditions, correlation coefficients (r) were >0.997 for calibration curves in the range 1-80 ng/mL, detection limits and quantification limits ranged from 0.19 to 0.67 and 0.63 to 2.23 ng/mL, respectively, and relative standard deviation was <8%. The method was applied for the determination of chlorophenols in real wines, with recovery rates in the range 82-104%. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Three dimensional liquid chromatography coupling ion exchange chromatography/hydrophobic interaction chromatography/reverse phase chromatography for effective protein separation in top-down proteomics.

    Science.gov (United States)

    Valeja, Santosh G; Xiu, Lichen; Gregorich, Zachery R; Guner, Huseyin; Jin, Song; Ge, Ying

    2015-01-01

    To address the complexity of the proteome in mass spectrometry (MS)-based top-down proteomics, multidimensional liquid chromatography (MDLC) strategies that can effectively separate proteins with high resolution and automation are highly desirable. Although various MDLC methods that can effectively separate peptides from protein digests exist, very few MDLC strategies, primarily consisting of 2DLC, are available for intact protein separation, which is insufficient to address the complexity of the proteome. We recently demonstrated that hydrophobic interaction chromatography (HIC) utilizing a MS-compatible salt can provide high resolution separation of intact proteins for top-down proteomics. Herein, we have developed a novel 3DLC strategy by coupling HIC with ion exchange chromatography (IEC) and reverse phase chromatography (RPC) for intact protein separation. We demonstrated that a 3D (IEC-HIC-RPC) approach greatly outperformed the conventional 2D IEC-RPC approach. For the same IEC fraction (out of 35 fractions) from a crude HEK 293 cell lysate, a total of 640 proteins were identified in the 3D approach (corresponding to 201 nonredundant proteins) as compared to 47 in the 2D approach, whereas simply prolonging the gradients in RPC in the 2D approach only led to minimal improvement in protein separation and identifications. Therefore, this novel 3DLC method has great potential for effective separation of intact proteins to achieve deep proteome coverage in top-down proteomics.

  2. Application of Genetic Algorithms for Parameter Estimation in Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Orestes Llanes Santiago

    2011-11-01

    Full Text Available Normal 0 21 false false false ES X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabla normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} In chromatography, complex inverse problems related to the parameters estimation and process optimization are presented. Metaheuristics methods are known as general purpose approximated algorithms which seek and hopefully find good solutions at a reasonable computational cost. These methods are iterative process to perform a robust search of a solution space. Genetic algorithms are optimization techniques based on the principles of genetics and natural selection. They have demonstrated very good performance as global optimizers in many types of applications, including inverse problems. In this work, the effectiveness of genetic algorithms is investigated to estimate parameters in liquid chromatography.

  3. Liquid chromatography-mass spectrometry in forensic toxicology.

    Science.gov (United States)

    Van Bocxlaer, J F; Clauwaert, K M; Lambert, W E; Deforce, D L; Van den Eeckhout, E G; De Leenheer, A P

    2000-01-01

    Liquid chromatography-mass spectrometry has evolved from a topic of mainly research interest into a routinely usable tool in various application fields. With the advent of new ionization approaches, especially atmospheric pressure, the technique has established itself firmly in many areas of research. Although many applications prove that LC-MS is a valuable complementary analytical tool to GC-MS and has the potential to largely extend the application field of mass spectrometry to hitherto "MS-phobic" molecules, we must recognize that the use of LC-MS in forensic toxicology remains relatively rare. This rarity is all the more surprising because forensic toxicologists find themselves often confronted with the daunting task of actually searching for evidence materials on a scientific basis without any indication of the direction in which to search. Through the years, mass spectrometry, mainly in the GC-MS form, has gained a leading role in the way such quandaries are tackled. The advent of robust, bioanalytically compatible combinations of liquid chromatographic separation with mass spectrometric detection really opens new perspectives in terms of mass spectrometric identification of difficult molecules (e.g., polar metabolites) or biopolymers with toxicological relevance, high throughput, and versatility. Of course, analytical toxicologists are generally mass spectrometry users rather than mass spectrometrists, and this difference certainly explains the slow start of LC-MS in this field. Nevertheless, some valuable applications have been published, and it seems that the introduction of the more universal atmospheric pressure ionization interfaces really has boosted interests. This review presents an overview of what has been realized in forensic toxicological LC-MS. After a short introduction into LC-MS interfacing operational characteristics (or limitations), it covers applications that range from illicit drugs to often abused prescription medicines and some

  4. Comparison of liquid and supercritical fluid chromatography mobile phases for enantioselective separations on polysaccharide stationary phases.

    Science.gov (United States)

    Khater, Syame; Lozac'h, Marie-Anne; Adam, Isabelle; Francotte, Eric; West, Caroline

    2016-10-07

    Analysis and production of enantiomerically pure compounds is a major topic of interest when active pharmaceutical ingredients are concerned. Enantioselective chromatography has become a favourite both at the analytical and preparative scales. High-performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) are dominating the scene and are often seen as complementary techniques. Nowadays, for economic and ecologic reasons, SFC may be preferred over normal-phase HPLC (NPLC) as it allows significant reductions in solvent consumption. However, the transfer of NPLC methods to SFC is not always straightforward. In this study, we compare the retention of achiral molecules and separation of enantiomers under supercritical fluid (carbon dioxide with ethanol or isopropanol) and liquid normal-phase (heptane with ethanol or isopropanol) elution modes with polysaccharide stationary phases in order to explore the differences between the retention and enantioseparation properties between the two modes. Chemometric methods (namely quantitative structure-retention relationships and discriminant analysis) are employed to compare the results obtained on a large set of analytes (171 achiral probes and 97 racemates) and gain some understanding on the retention and separation mechanisms. The results indicate that, contrary to popular belief, carbon dioxide - solvent SFC mobile phases are often weaker eluents than liquid mobile phases. It appears that SFC and NPLC elution modes provide different retention mechanisms. While some enantioseparations are unaffected, facilitating the transfer between the two elution modes, other enantioseparations may be drastically different due to different types and strength of interactions contributing to enantioselectivity. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Integrated liquid chromatography-heated nebulizer microchip for mass spectrometry.

    Science.gov (United States)

    Haapala, Markus; Saarela, Ville; Pól, Jaroslav; Kolari, Kai; Kotiaho, Tapio; Franssila, Sami; Kostiainen, Risto

    2010-03-10

    A new integrated microchip for liquid chromatography-mass spectrometry (LC-MS) is presented. The chip is made from bonded silicon and glass wafers with structures for a packed LC column channel, a micropillar frit, a channel for optional optical detection, and a heated vaporizer section etched in silicon and platinum heater elements on the glass cover. LC eluent is vaporized and mixed with nebulizer gas in the vaporizer section and the vapor is sprayed out from the chip. Nonpolar and polar analytes can be efficiently ionized in the gas phase by atmospheric pressure photoionization (APPI) as demonstrated with polycyclic aromatic hydrocarbons (PAHs) and selective androgen receptor modulators (SARMs). This is not achievable with present LC-MS chips, since they are based on electrospray ionization, which is not able to ionize nonpolar compounds efficiently. The preliminary quantitative performance of the new chip was evaluated in terms of limit of detection (down to 5 ng mL(-1)), linearity (r>0.999), and repeatability of signal response (RSD=2.6-4.0%) and retention time (RSD=0.3-0.5%) using APPI for ionization and PAHs as standard compounds. Determination of fluorescent compounds is demonstrated by using laser-induced fluorescence (LIF) for detection in the optical detection channel before the vaporizer section. 2010 Elsevier B.V. All rights reserved.

  6. Determination of saffron quality by high-performance liquid chromatography.

    Science.gov (United States)

    Valle García-Rodríguez, M; Serrano-Díaz, Jéssica; Tarantilis, Petros A; López-Córcoles, Horacio; Carmona, Manuel; Alonso, Gonzalo L

    2014-08-13

    The aim of this work was to propose a high-performance liquid chromatography with diode array detection (HPLC-DAD) method for determining the three main compounds responsible for determining the quality of saffron (crocetin esters, picrocrocin, and safranal) by preparing an aqueous extract according to the ISO 3632 standard to solve the difficulty that this standard has for aroma and taste determination by ultraviolet-visible spectroscopy. Toward this aim, laboratory-isolated picrocrocin, a safranal standard with a purity of ≥ 88%, trans-crocetin di(β-D-gentiobiosyl) ester (trans-4-GG) and trans-crocetin (β-D-glucosyl)-(β-D-gentiobiosyl) ester (trans-3-Gg) standards, both with a purity of ≥ 99%, and 50 different saffron spice samples from Italy, Iran, Greece, and Spain were used in the intralaboratory validation of the HPLC method. The analytical method proposed was adequate in terms of linearity, selectivity, sensitivity, and accuracy for determining the three foremost parameters that define the quality of saffron using only a saffron solution prepared according to the ISO 3632 standard.

  7. Characterization of porous silicon integrated in liquid chromatography chips.

    Science.gov (United States)

    Tiggelaar, R M; Verdoold, V; Eghbali, H; Desmet, G; Gardeniers, J G E

    2009-02-07

    Properties of porous silicon which are relevant for use of the material as a stationary phase in liquid chromatography chips, like porosity, pore size and specific surface area, were determined with high-resolution SEM and N(2) adsorption-desorption isotherms. For the anodization conditions investigated, porosity is between 20 and 60%, pore sizes between 2 and 5 nm and specific surface area between 130 and 410 m(2)/cm(3). It was established that under identical anodization conditions, porous layer formation is 10-15% slower on micromachined pillars than on flat substrates, and depends on geometrical parameters like pillar diameter and height and interpillar spacing. In microchannels containing pillars with a porous silicon shell, chromatographic experiments on a coumarin dye mixture were performed, which in comparison with non-porous pillars showed a significant increase of the retention factors, resulting from the large internal surface of the porous pillars. The increased relative retention of one of the coumarin dyes, C480, could be correlated quantitatively with the measured internal surface of the porous layer. Due to the small pore size, these porous shell columns are particularly suitable for analytical or preparative separation of low-molecular weight molecules, with applications in metabolomics, food quality control, or medical diagnostics.

  8. Strategies for metabolite profiling based on liquid chromatography.

    Science.gov (United States)

    Saurina, Javier; Sentellas, Sonia

    2017-02-15

    This paper aims at covering the principal strategies based on liquid chromatography (LC) for metabolite profiling in the field of drug discovery and development. The identification of metabolites generated in the organism is an important task during the early stages of preclinical research to define the most proper strategy for optimizing, adjusting metabolic clearance and minimizing bioactivation. An early assessment of the metabolite profile may be critical since metabolites can contribute to pharmacological and/or toxicological effects. The study of metabolites first involves their synthesis/generation and their further characterization and structural elucidation. For such a purpose, both in vitro and in vivo methods are commonly used for the generation of the corresponding metabolites. Next, analytical methods are used to tackle identification and characterization studies. Among the arsenal of techniques available in our labs, we will focus on LC, especially coupled to mass spectrometry (LC-MS), as one of the most powerful approaches for metabolite identification, characterization and quantification. Here, the topic of metabolite profiling based on LC will be addressed and representative examples of different possibilities will be discussed. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Determination of carotenoids in tomato juice by liquid chromatography.

    Science.gov (United States)

    Lin, C H; Chen, B H

    2003-09-12

    A high-performance liquid chromatography method was developed to determine the various carotenoids in tomato juice. A C30 column and a mobile phase of acetonitrile-1-butanol (7:3, v/v) (A) and methylene chloride (B) with the following gradient elution were used: 99% A and 1% B intitally, increased to 4% B in 20 min, 10% B in 50 min and returned to 1% B in 55 min. Sixteen carotenoids, including all-trans-lutein, all-trans-beta-carotene, all-trans-lycopene and their 13 cis isomers were identified and resolved within 52 min with flow-rate at 2.0 ml/min and detection at 476 nm. Of the various extraction solvent systems, the best extraction efficiency of carotenoids in tomato juice was achieved by employing ethanol-hexane (4:3, v/v). Lycopene was found to be present in largest amount in tomato juice, followed by beta-carotene and lutein.

  10. Investigation of Symphytum cordatum alkaloids by liquid-liquid partitioning, thin-layer chromatography and liquid chromatography-ion-trap mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Mroczek, Tomasz [Department of Pharmacognosy with Medicinal Plants Laboratory, Medical University, 1 Chodzki St., 20-093 Lublin (Poland)]. E-mail: tmroczek@pharmacognosy.org; Ndjoko-Ioset, Karine [Laboratoire de Pharmacognosie et Phytochimie, Ecole de Pharmacie Geneve-Lausanne, Universite de Geneve, Quai Ernest-Ansermet 30, CH-1211 Geneva 4 (Switzerland); Glowniak, Kazimierz [Department of Pharmacognosy with Medicinal Plants Laboratory, Medical University, 1 Chodzki St., 20-093 Lublin (Poland); Mietkiewicz-Capala, Agnieszka [Department of Pharmacognosy with Medicinal Plants Laboratory, Medical University, 1 Chodzki St., 20-093 Lublin (Poland); Hostettmann, Kurt [Laboratoire de Pharmacognosie et Phytochimie, Ecole de Pharmacie Geneve-Lausanne, Universite de Geneve, Quai Ernest-Ansermet 30, CH-1211 Geneva 4 (Switzerland)

    2006-05-04

    From the alkalised crude extract of Symphytum cordatum (L.) W.K. roots, pyrrolizidine alkaloids (PAs) were extracted as free tertiary bases and polar N-oxides in a merely one-step liquid-liquid partitioning (LLP) in separation funnel and subsequently pre-fractionated by preparative multiple-development (MD) thin-layer chromatography (TLC) on silica gel plates. In this way three alkaloid fractions of different polarities and retention on silica gel plates were obtained as: the most polar N-oxides of the highest retention, the tertiary bases of medium retention, and diesterified N-oxides of the lowest retention. The former fraction was reduced into free bases by sodium hydrosulfite and purified by LLP on Extrelut-NT3 cartridge. It was further analysed together with the two other fractions by high-performance liquid chromatography (HPLC)-ion-trap mass spectrometry with atmospheric pressure chemical ionization (APCI) interface on XTerra C{sub 18} column using a gradient elution. Based on MS {sup n} spectra, 18 various alkaloids have been tentatively determined for the first time in this plant as the following types of structure: echimidine-N-oxide (three diasteroisomers), 7-sarracinyl-9-viridiflorylretronecine (two diasteroisomers), echimidine (two diasteroisomers), lycopsamine (two diasteroisomers), dihydroechinatine-N-oxide, dihydroheliospathuline-N-oxide, lycopsamine-N-oxide (three diasteroisomers), 7-acetyllycopsamine-N-oxide, symphytine-N-oxide (two diasteroisomers) and 2'',3''-epoxyechiumine-N-oxide.

  11. Flexible and Accessible Automated Operation of Miniature Chromatography Columns on a Liquid Handling Station.

    Science.gov (United States)

    Konstantinidis, Spyridon; Goh, Hai-Yuan; Martin Bufájer, José M; de Galbert, Paul; Parau, Maria; Velayudhan, Ajoy

    2017-11-13

    The High Throughput (HT) investigation of chromatographic separations is an important element of downstream bioprocess development due to the importance of chromatography as a technique for achieving stringent regulatory requirements on product purity. Various HT formats for chromatography exist, but the miniature column approach has characteristics resembling large scale packed bed column chromatography the most. The operation of such columns on robotic stations can be automated, but this is not always a straightforward procedure; the robotic manipulations are highly dependent on the settings of each experiment and the standard commands of the supporting software may not provide readily the required flexibility and accessibility for "plug and play" functionality. These can limit the potential of this technique in laboratories engaging on HT activities. In this work, we present an application which aims to overcome this challenge by providing end-users with a flexible operation of the miniature column technique on an automated liquid handler. The application includes a script which is written on Freedom EVOware, and is supplemented by custom compiled executables. Here, the manipulations carried out by the application are described in detail and its functionality is demonstrated through typical experiments based on bind and elute miniature column chromatography. The application is shown to allow for the unsupervised "on-the-fly" programming of the robotic station and to ultimately make the technique accessible to non-automation experts. This application is therefore well suited to simplifying development activities based on the robotic deployment of the miniature column chromatography technique. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Countercurrent tangential chromatography for large-scale protein purification.

    Science.gov (United States)

    Shinkazh, Oleg; Kanani, Dharmesh; Barth, Morgan; Long, Matthew; Hussain, Daniar; Zydney, Andrew L

    2011-03-01

    Recent advances in cell culture technology have created significant pressure on the downstream purification process, leading to a "downstream bottleneck" in the production of recombinant therapeutic proteins for the treatment of cancer, genetic disorders, and cardiovascular disease. Countercurrent tangential chromatography overcomes many of the limitations of conventional column chromatography by having the resin (in the form of a slurry) flow through a series of static mixers and hollow fiber membrane modules. The buffers used in the binding, washing, and elution steps flow countercurrent to the resin, enabling high-resolution separations while reducing the amount of buffer needed for protein purification. The results obtained in this study provide the first experimental demonstration of the feasibility of using countercurrent tangential chromatography for the separation of a model protein mixture containing bovine serum albumin and myoglobin using a commercially available anion exchange resin. Batch uptake/desorption experiments were used in combination with critical flux data for the hollow fiber filters to design the countercurrent tangential chromatography system. A two-stage batch separation yielded the purified target protein at >99% purity with 94% recovery. The results clearly demonstrate the potential of using countercurrent tangential chromatography for the large-scale purification of therapeutic proteins. Copyright © 2010 Wiley Periodicals, Inc.

  13. Simultaneous analysis for water- and fat-soluble vitamins by a novel single chromatography technique unifying supercritical fluid chromatography and liquid chromatography.

    Science.gov (United States)

    Taguchi, Kaori; Fukusaki, Eiichiro; Bamba, Takeshi

    2014-10-03

    Chromatography techniques usually use a single state in the mobile phase, such as liquid, gas, or supercritical fluid. Chromatographers manage one of these techniques for their purpose but are sometimes required to use multiple methods, or even worse, multiple techniques when the target compounds have a wide range of chemical properties. To overcome this challenge, we developed a single method covering a diverse compound range by means of a "unified" chromatography which completely bridges supercritical fluid chromatography and liquid chromatography. In our method, the phase state was continuously changed in the following order; supercritical, subcritical and liquid. Moreover, the gradient of the mobile phase starting at almost 100% CO2 was replaced with 100% methanol at the end completely. As a result, this approach achieved further extension of the polarity range of the mobile phase in a single run, and successfully enabled the simultaneous analysis of fat- and water-soluble vitamins with a wide logP range of -2.11 to 10.12. Furthermore, the 17 vitamins were exceptionally separated in 4min. Our results indicated that the use of dense CO2 and the replacement of CO2 by methanol are practical approaches in unified chromatography covering diverse compounds. Additionally, this is a first report to apply the novel approach to unified chromatography, and can open another door for diverse compound analysis in a single chromatographic technique with single injection, single column and single system. Copyright © 2014. Published by Elsevier B.V.

  14. Electrochemically modulated liquid chromatography: Theoretical investigations and applications from the perspectives of chromatography and interfacial electrochemistry

    Energy Technology Data Exchange (ETDEWEB)

    Keller, David W. [Iowa State Univ., Ames, IA (United States)

    2005-01-01

    Electrochemically modulated liquid chromatography (EMLC) employs a conductive material as both a stationary phase for chromatographic separations and as a working electrode for performing electrochemistry experiments. This dual functionality gives EMLC the capacity to manipulate chromatographic separations by changing the potential applied (Eapp) to the stationary phase with respect to an external reference. The ability to monitor retention as a function of Eapp provides a means to chromatographically monitor electrosorption processes at solid-liquid interfaces. In this dissertation, the retention mechanism for EMLC is examined from the perspective of electrical double layer theory and interfacial thermodynamics. From the chromatographic data, it is possible to determine the interfacial excess (Λ) of a solute and changes in interfacial tension (dγ) as a function of both Eapp and the supporting electrolyte concentration. Taken together, these two experimentally manipulated parameters can be examined within the context of the Gibbs adsorption equation to delineate the contribution of a variety of interfacial properties, including the charge of solute on the stationary phase and the potential of zero charge (PZC), to the mechanism behind EMLC-based retention. The chromatographic probing of interfacial phenomena is complemented by electroanalytical experiments that exploit the ability to monitor the electronic current flowing through an EMLC column. Cyclic voltammetry and chronoamperometry of an EMLC column are used to determine the electronic performance characteristics of an EMLC column. An electrochemical flow injection analysis of a column is provided in which the current required to maintain a constant Eapp is monitored and provides a way to examine the influence that acetonitrile and supporting electrolyte composition, flow rate, column backpressure, and ionic strength have on the structure of electrified interfaces.

  15. Liquid chromatography-mass spectrometry coupling by the intermediary of a liquid micro chromatography-electro spray interface; Couplage chromatographie liquide-spectrometrie de masse par l`intermediaire d`une interface electrospray-microchromatographie liquide

    Energy Technology Data Exchange (ETDEWEB)

    Gillard Factor, C.

    1996-12-06

    The objective of this work is to realize a liquid chromatography- mass spectrometry coupling by the intermediary of an electro spray interface and the evaluation of performances of tis analytical tool to study pollutants in water, and more particularly pesticides whom maximum admissible concentration in a table water is 0.1{mu}g/l. This study has allowed to bring to the fore the interest of the ionization mode by electro spray in a LC/MS coupling to identify and quantify pesticides in the state of traces without treating the sample. Then, it was demonstrated the usefulness of this analytical tool to detect high molecular masses molecules. (N.C.)

  16. New methods and materials for solid phase extraction and high performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Dumont, Philip John [Iowa State Univ., Ames, IA (United States)

    1996-04-23

    This paper describes methods for solid phase extraction and high performance liquid chromatography (HPLC). The following are described: Effects of Resin Sulfonation on the Retention of Polar Organic Compounds in Solid Phase Extraction; Ion-Chromatographic Separation of Alkali Metals In Non-Aqueous Solvents; Cation-Exchange Chromatography in Non-Aqueous Solvents; and Silicalite As a Stationary Phase For HPLC.

  17. Reducing Dilution and Analysis Time in Online Comprehensive Two-Dimensional Liquid Chromatography by Active Modulation

    NARCIS (Netherlands)

    Gargano, A.F.G.; Duffin, M.; Navarro, P.; Schoenmakers, P.J.

    2016-01-01

    Online comprehensive two-dimensional liquid chromatography (LC × LC) offers ways to achieve high-performance separations in terms of peak capacity (exceeding 1000) and additional selectivity to realize applications that cannot be addressed with one-dimensional chromatography (1D-LC). However, the

  18. Liquid chromatography-tandem mass spectrometry - Application in the clinical laboratory

    OpenAIRE

    Vogeser, Michael

    2003-01-01

    This review provides a concise survey of liquid chromatography tandem mass spectrometry (LCTMS) as an emerging technology in clinical chemistry. The combination of two mass spectrometers with an interposed collision cell characterizes LCTMS as an analytical technology on its own and not just as a more specific detector for HPLC compared with conventional techniques. In LCTMS, liquid chromatography is rather used for sample preparation but not for complete resolution of compounds of interest. ...

  19. Tryptophan and kynurenine determination in human hair by liquid chromatography.

    Science.gov (United States)

    Dario, Michelli F; Freire, Thamires Batello; Pinto, Claudinéia Aparecida Sales de Oliveira; Prado, María Segunda Aurora; Baby, André R; Velasco, Maria Valéria R

    2017-10-15

    Tryptophan, an amino acid found in hair proteinaceous structure is used as a marker of hair photodegradation. Also, protein loss caused by several chemical/physical treatments can be inferred by tryptophan quantification. Kynurenine is a photo-oxidation product of tryptophan, expected to be detected when hair is exposed mainly to UVB (290-320nm) radiation range. Tryptophan from hair is usually quantified directly as a solid or after alkaline hydrolysis, spectrofluorimetrically. However, these types of measure are not sufficiently specific and present several interfering substances. Thus, this work aimed to propose a quantification method for both tryptophan and kynurenine in hair samples, after alkali hydrolysis process, by using high-performance liquid chromatography (HPLC) with fluorimetric and UV detection. The tryptophan and kynurenine quantification method was developed and validated. Black, white, bleached and dyed (blond and auburn) hair tresses were used in this study. Tryptophan and kynurenine were separated within ∼9min by HPLC. Both black and white virgin hair samples presented similar concentrations of tryptophan, while bleaching caused a reduction in the tryptophan content as well as dyeing process. Unexpectedly, UV/vis radiation did not promote significantly the conversion of tryptophan into its photo-oxidation product and consequently, kynurenine was not detected. Thus, this works presented an acceptable method for quantification of tryptophan and its photooxidation metabolite kynurenine in hair samples. Also, the results indicated that bleaching and dyeing processes promoted protein/amino acids loss but tryptophan is not extensively degraded in human hair by solar radiation. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Liquid chromatography method to determine polyamines in thermosetting polymers

    Energy Technology Data Exchange (ETDEWEB)

    Dopico-Garcia, M.S. [Laboratorio de Quimica - Centro de Investigacions Tecnoloxicas, Universidade da Coruna, Campus de Esteiro s/n, 15403 Ferrol (Spain); Centro Galego do Plastico, A Cabana s/n, 15590 Ferrol (Spain); Lopez-Vilarino, J.M. [Laboratorio de Quimica - Centro de Investigacions Tecnoloxicas, Universidade da Coruna, Campus de Esteiro s/n, 15403 Ferrol (Spain); Fernandez-Martinez, G. [Unidad de Tecnicas Cromatograficas, Servizos de Apoio a Investigacion, Edificio Servizos Centrais de Investigacion, Universidade da Coruna, Campus de Elvina s/n, 15071 A Coruna (Spain); Gonzalez-Rodriguez, M.V., E-mail: victoria@udc.es [Dpto. de Quimica Analitica - E.U. Politecnica, Universidade da Coruna, Avda. 19 de Febrero s/n, 15405 Ferrol (Spain)

    2010-05-14

    A simple, robust and sensitive analytical method to determine three polyamines commonly used as hardeners in epoxy resin systems and in the manufacture of polyurethane is reported. The studied polyamines are: one tetramine, TETA (triethylenetetramine), and two diamines, IPDA (Isophorone diamine) and TCD-diamine (4,7-methano-1H-indene-5,?-dimethanamine, octahydro-). The latter has an incompletely defined structure, and, as far as we know, has not been previously determined by other methods. All three polyamines contain primary amines; TETA also contains secondary amines. The analytical method involves derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, used for the first time for these compounds, followed by high performance liquid chromatography (HPLC) analysis with a fluorescence (FL) detector ({lambda} excitation 248 nm, {lambda} emision 395 nm). The HPLC-DAD-LTQ Orbitrap MS was used in order to provide structural information about the obtained derivatized compounds. The hybrid linear ion trap LTQ Orbitrap mass spectrometer has been introduced in recent years and provides a high mass accuracy. The structures of the derivatized analytes were identified from the protonated molecular ions [M+H]{sup +} and corresponded to the fully labelled analytes. The following analytical parameters were determined for the method using the HPLC-FL: linearity, precision (2.5-10%), instrumental precision intraday (0.8-1.5%) and interday (2.9-6.3%), and detection limits (0.02-0.14 mg L{sup -1}). The stability of stock solutions and derivatized compounds was also investigated. The method was applied to determine the amine free content in epoxy resin dust collected in workplaces.

  1. Development of an on-line mixed-mode gel liquid chromatography×reversed phase liquid chromatography method for separation of water extract from Flos Carthami.

    Science.gov (United States)

    Wang, Yu-Qing; Tang, Xu; Li, Jia-Fu; Wu, Yun-Long; Sun, Yu-Ying; Fang, Mei-Juan; Wu, Zhen; Wang, Xiu-Min; Qiu, Ying-Kun

    2017-10-13

    A novel on-line comprehensive two-dimensional liquid chromatography (2D-LC) method by coupling mixed-mode gel liquid chromatography (MMG-LC) with reversed phase liquid chromatography (RPLC) was developed. A mixture of 17 reference compounds was used to study the separation mechanism. A crude water extract of Flos Carthami was applied to evaluate the performance of the novel 2D-LC system. In the first dimension, the extract was eluted with a gradient of water/methanol over a cross-linked dextran gel Sephadex LH-20 column. Meanwhile, the advantages of size exclusion, reversed phase partition and adsorption separation mechanism were exploited before further on-line reversed phase purification on the second dimension. This novel on-line mixed-mode Sephadex LH-20×RPLC method provided higher peak resolution, sample processing ability (2.5mg) and better orthogonality (72.9%) versus RPLC×RPLC and hydrophilic interaction liquid chromatography (HILIC)×RPLC. To the best of our knowledge, this is the first report of a mixed-mode Sephadex LH-20×RPLC separation method with successful applications in on-line mode, which might be beneficial for harvesting targets from complicated medicinal plants. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Evaluation of a silicon oxynitride hydrophilic interaction liquid chromatography column in saccharide and glycoside separations.

    Science.gov (United States)

    Wan, Huihui; Sheng, Qianying; Zhong, Hongmin; Guo, Xiujie; Fu, Qing; Liu, Yanfang; Xue, Xingya; Liang, Xinmiao

    2015-05-01

    The retention characteristics of a silicon oxynitride stationary phase for carbohydrate separation were studied in hydrophilic interaction chromatography mode. Four saccharides including mono-, di-, and trisaccharides were employed to investigate the effects of water content and buffer concentration in the mobile phase on hydrophilic interaction liquid chromatography retention. For the tested saccharides, the silicon oxynitride column demonstrated excellent performance in terms of separation efficiency, hydrophilicity, and interesting separation selectivity for carbohydrates compared to the bare silica stationary phase. Finally, the silicon oxynitride hydrophilic interaction liquid chromatography column was employed in the separation of complex samples of fructooligosaccharides, saponins, and steviol glycoside from natural products. The resulting chromatograms demonstrated good separation efficiency and longer retention compared with silica, which further confirmed the advantages and potential application of silicon oxynitride stationary phase for hydrophilic interaction liquid chromatography separation. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Analysis of Peroxidases Using Liquid Chromatography, Post-Column Substrate Conversion and Fluorescence Detection

    NARCIS (Netherlands)

    Haselberg, Rob; Hempen, C.M.; van Leeuwen, S.M.; Vogel, M.; Karst, U.

    2006-01-01

    A liquid chromatographic method with on-line activity determination for microperoxidases has been developed. After enzymatic digestion of a cytochrome, possibly under formation of microperoxidases, the product mixture is separated by reversed-phase liquid chromatography. The products first pass a

  4. STRUCTURAL CHARACTERIZATION OF THE DECOMPOSITION PRODUCTS OF SALBUTAMOL BY LIQUID-CHROMATOGRAPHY IONSPRAY MASS-SPECTROMETRY

    NARCIS (Netherlands)

    MALKKILAINE, L; BRUINS, AP

    Liquid chromatography-ionspray mass spectrometry was used to elucidate the structures of the decomposition products of salbutamol. The best sensitivity in mass spectrometry was achieved by using a mixture of acetonitrile and ammonium formate (10 mM, pH 3.3) as the mobile phase in liquid

  5. Denaturing high performance liquid chromatography (DHPLC in detection of microsatellite instability

    Directory of Open Access Journals (Sweden)

    Metka Ravnik-Glavač

    2007-12-01

    Full Text Available Background: Microsatellite instability (MSI is a phenomenon characterized by small deletions or insertions within short tandem repeats in tumour DNA compared to matching normal DNA. MSI analysis is becoming more and more important for detection of hereditary non-polyposis colorectal cancer as well as for sporadic primary colorectal tumours with MSI high phenotype. Use of five quasimonomorphic mononucleotide markers eliminates ultimate need for analysis of germline DNA corresponding to tumour DNA. Here we discuss our method for MSI analysis using denaturating high performance liquid chromatography (DHPLC in combination with quasimonomorphic mononucleotide microsatellite markers in comparison with previously used methods. The method is high-throughput, accurate, quick and cost-effective and suitable for large-scale studies as well as for daily use with smaller numbers of samples.

  6. Chromatography.

    Science.gov (United States)

    Brantley, L. Reed, Sr.; Demanche, Edna L.; Klemm, E. Barbara; Kyselka, Will; Phillips, Edwin A.; Pottenger, Francis M.; Yamamoto, Karen N.; Young, Donald B.

    This booklet presents some activities on chromatography. Directions for preparing leaf pigment extracts using alcohol are given, and paper chromatography and thin-layer chromatography are described as modifications of the basic principles of chromatography. (KHR)

  7. Potentiometric detection of organic acids in liquid chromatography using polymeric liquid membrane electrodes incorporating macrocyclic hexaamines.

    Science.gov (United States)

    Zielinska, D; Poels, I; Pietraszkiewicz, M; Radecki, J; Geise, H J; Nagels, L J

    2001-04-27

    Potentiometric detection employing coated-wire electrodes was applied to the determination of organic acids in liquid chromatography (LC). Poly(vinyl chloride)-based liquid membranes, incorporating lipophilic macrocyclic hexaamines as neutral ionophores were used as electrode coatings. The selectivity and sensitivity of the macrocycle-based electrodes were found to be superior to an electrode based on a lipophilic anion exchanger (a quaternary ammonium salt). Sensitive detection was obtained for the di- and tricarboxylic acids tartaric, malonic, malic, citric, fumaric, succinic, pyruvic, 2-oxoglutaric and maleic acids after separation in reversed-phase LC. Detection limits (signal/4sigmanoise=3) of 6 pmol for malonic acid and 2 pmol for maleic acid were attained. The detection was explained using a molecular recognition model. The hexaamine-based potentiometric electrodes had a 1-s response time at 1 ml min(-1) flow-rates. They were stable for at least 4 months, with an intra-electrode variation of 3.2% (n=5).

  8. Characterization of synthetic dyes by comprehensive two-dimensional liquid chromatography combining ion-exchange chromatography and fast ion-pair reversed-phase chromatography.

    Science.gov (United States)

    Pirok, Bob W J; Knip, Jitske; van Bommel, Maarten R; Schoenmakers, Peter J

    2016-03-04

    In the late 19th century, newly invented synthetic dyes rapidly replaced the natural dyes on the market. The characterization of mixtures of these so-called early synthetic dyes is complicated through the occurrence of many impurities and degradation products. Conventional one-dimensional liquid chromatography does not suffice to obtain fingerprints with sufficient resolution and baseline integrity. Comprehensive two-dimensional liquid chromatography (LC×LC) is employed in this study, with ion-exchange chromatography in the first dimension and fast ion-pair liquid chromatography in the second. Retention in the first dimension is largely determined by the number of charges, while the selection of a small ion-pair reagent (tetramethylammonium hydroxide) in the second dimension causes retention to be largely determined by the molecular structure of the dye. As a result, there is a high degree of orthogonality of the two dimensions, similar to the values typically encountered in GC×GC. The proposed LC×LC method shows a theroretical peak capacity of about 2000 in an analysis time of about three hours. Clear, informative fingerprints are obtained that open a way to a more efficient characterization of dyes used in objects of cultural heritage. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Screening and quantitative determination of twelve acidic and neutral pharmaceuticals in whole blood by liquid-liquid extraction and liquid chromatography-tandem mass spectrometry

    DEFF Research Database (Denmark)

    Simonsen, Kirsten Wiese; Steentoft, Anni; Buck, Maike

    2010-01-01

    . The method was fully validated for salicylic acid, paracetamol, phenobarbital, carisoprodol, meprobamate, topiramate, etodolac, chlorzoxazone, furosemide, ibuprofen, warfarin, and salicylamide. The method also tentatively includes thiopental, theophylline, piroxicam, naproxen, diclophenac, and modafinil......We describe a multi-method for simultaneous identification and quantification of 12 acidic and neutral compounds in whole blood. The method involves a simple liquid-liquid extraction, and the identification and quantification are performed using liquid chromatography-tandem mass spectrometry...

  10. Improved method for the determination of the cortisol production rate using high-performance liquid chromatography and liquid scintillation counting

    NARCIS (Netherlands)

    van Ingen, H. E.; Endert, E.

    1988-01-01

    Two new methods for the determination of the cortisol production rate using reversed-phase high-performance liquid chromatography are described. One uses ultraviolet detection at 205 nm, the other on-line post-column derivatization with benzamidine, followed by fluorimetric detection. The specific

  11. Rapid, simple and stability-indicating determination of polyhexamethylene biguanide in liquid and gel-like dosage forms by liquid chromatography with diode-array detection

    National Research Council Canada - National Science Library

    Küsters, Markus; Beyer, Sören; Kutscher, Stephan; Schlesinger, Harald; Gerhartz, Michael

    2013-01-01

    A rapid and simple method for the determination of potyhexamethylene biguanide (polyhexanide, PHMB) in liquid and gel-like pharmaceutical formulations by means of high performance liquid chromatography coupled to diode-array detection...

  12. Evaluation of nanoflow liquid chromatography high resolution mass spectrometry for pesticide residue analysis in food.

    Science.gov (United States)

    Moreno-González, David; Pérez-Ortega, Patricia; Gilbert-López, Bienvenida; Molina-Díaz, Antonio; García-Reyes, Juan F; Fernández-Alba, Amadeo R

    2017-08-25

    This article reports on the evaluation of nanoflow liquid chromatography-mass spectrometry (LC-MS) for pesticide residue analysis in food. The approach is based on the use of reversed-phase C18nano columns with an integrated emitter, so that separation, ionization and detection are performed minimizing dead volumes. The use of nanoflow not only increases ionization efficiency and minimizes ionization suppression but also boost sensitivity compared to analytical-scale LC-MS methods. The nanoflow LC system was combined with full-scan high resolution mass spectrometry using a Q-Exactive Orbitrap instrument. The analytical performance was assessed for over 60 representative pesticides in five representative commodities (tomato, baby food, orange, fruit-based jam and olive oil). The sensitivity achieved with this configuration enables the implementation of high dilution factors (eg. 1:20, 1:50 or beyond) in pesticide residue workflows without compromising sensitivity, featuring limits of quantitation in the low ng kg-1 range. Using this dilution factors, signal suppression was found negligible in most cases (<10% in most cases, especially with 1:50 dilution), so that matrix-matched standards may be skipped, thus simplifying laboratory workflows. The robustness of the nanoflow LC system and its capability to withstand long analytical runs was also evaluated. Appropriate precision in terms of peak area and retention time was obtained at different concentration levels for over 125 injections without any instrument servicing. The main benefits of the nanoflow liquid chromatography approach are the high sensitivity gain and the outstanding reduction in matrix effects thanks to the high sample dilution factors that can be implemented, along with the substantial reduction in solvent usage. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Analytical confirmation of lethal heroin overdose by the use of liquid chromatography methods

    Directory of Open Access Journals (Sweden)

    Đorđević Snežana

    2007-01-01

    Full Text Available Background/Aim. Heroin is diacetylated morphine. Its ability to induce euphoria has led to its frequent abuse, giving rise to psychological and physical dependence. It has a short half-life, of approximately 2−6 min. In the brain, heroin undergoes deacetylation to 6-monoacetylmorphine (6−MAM and morphine. Detection of 6-acetylmorphine in the urine is indicative of heroin use. The aim of this study was to compare sensitivity and reliability of two analytical methods, a multicolumn liquid chromatography system with UV scanning detector (HPLCUV and liquid chromatography-mass spectrometry detection (LC-MS in opiate determining in post mortem material. Methods. Post mortem samples (blood, urine and vitreous humor were analyzed by liquid chromatography with UV and MS detection. The samples were prepared by liquid-liquid extraction with mixture chloroform-isopropanol (9:1. Separation was performed on C8 column with mobile phase composed of 55% acetonitrile-glacial acetic acid (99:1 and 45% 20 mM ammonium acetate. Results. The analysis of blood samples, urine, and eye liquid by the use of multicolumn HPLC-UV method confirmed the presence of morphine in the samples of blood and urine, codeine only in urine, and 6-MAM in the samples of urine and eye liquid. Using LC-MS method morphine was confirmed in all of the samples, while codeine was confirmed in urine and in the sample of eye liquid. In the samples of eye liquid and urine 6-MAM was confirmed. Conclusion. For determination of opiates in post mortem material LC-MS technique is more sensitive and reliable as compared to multicolumn liquid chromatography.

  14. Separation of cashew nut shell liquid by column chromatography ...

    African Journals Online (AJOL)

    compounds was separated into cardanol, cardol, and 2 - methylcardol using column chromatography. The separation was aimed at recovering cardanol that can be used in the synthesis of cation-exchange resin. The separation was effected using a mixture of equal portions of benzene and chloroform as the mobile phase ...

  15. Monodisperse microbeads of hypercrosslinked polystyrene for liquid and supercritical fluid chromatography

    Science.gov (United States)

    Tsyurupa, M. P.; Blinnikova, Z. K.; Il'in, M. M.; Davankov, V. A.; Parenago, O. O.; Pokrovskii, O. I.; Usovich, O. I.

    2015-11-01

    Monodisperse styrene-divinylbenzene (1 wt %) copolymer microbeads are obtained via the elaborate method of high-productivity precipitation polymerization. The crosslinking of this copolymer with chloromethyl methyl ether in the presence of Friedel-Crafts catalyst yields porous hypercrosslinked polymers with degrees of crosslinking that range from 200 to 500%. Microbead sorbents are shown to be suited for selective stationary phases for high-performance liquid chromatography and supercritical fluid chromatography.

  16. Determination of organophosphorus pesticides in soil by dispersive liquid-liquid microextraction and gas chromatography.

    Science.gov (United States)

    Yang, Zhonghua; Liu, Yu; Liu, Donghui; Zhou, Zhiqiang

    2012-01-01

    In this article, a rapid and sensitive sample pretreatment technique for the determination of organophosphorus pesticides (OPPs) in soil samples is developed by using dispersive liquid-liquid microextraction (DLLME) combined with gas chromatography-flame photometric detection. Experimental conditions, including the kind of extraction and disperser solvent and their volumes, the extraction time, and the salt addition, are investigated, and the following experiment factors are used: 20 µL chlorobenzene as the extraction solvent; 1.0 mL acetonitrile as the disperser solvent; no addition of salt; and an extraction time of 1 min. Under the optimum conditions, the linearities for the three target OPPs (ethoprophos, chlorpyriphos, and profenofos) are obtained by five points in the concentration range of 2.5-1500 µg/kg, and three replicates are used for each point. Correlation coefficients vary from 0.9987 to 0.9997. The repeatability is tested by spiking soil samples at a concentration level of 5.0 µg/kg. The relative standard deviation (n = 3) varied between 2.0% and 6.6%. The limits of detection, based on a signal-to-noise ratio (S/N) of 3, range from 200 to 500 pg/g. This method is applied to the analysis of the spiked samples S1, S2, and S3, which are collected from the China Agriculture University's orchard, lawn, and garden, respectively. The recoveries for each target analyte are in the range between 87.9% and 108.0%, 87.4% and 108.0%, and 86.7% and 107.2%, respectively. © The Author [2011]. Published by Oxford University Press. All rights reserved.

  17. Determination of muscimol and ibotenic acid in Amanita mushrooms by high-performance liquid chromatography and liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Tsujikawa, Kenji; Kuwayama, Kenji; Miyaguchi, Hajime; Kanamori, Tatsuyuki; Iwata, Yuko; Inoue, Hiroyuki; Yoshida, Takemi; Kishi, Tohru

    2007-06-01

    A reliable analytical method was developed for the quantification and identification of muscimol (MUS) and ibotenic acid (IBO), the toxic constituents of Amanita muscaria and Amanita pantherina. MUS and IBO were extracted from mushrooms by aqueous methanol and derivatized with dansyl chloride (DNS-Cl). After extraction with ethyl acetate and evaporation of the solvent, the residue was ethylated with 1.25 M hydrogen chloride in ethanol. The resulting derivatives were quantified by high-performance liquid chromatography with UV detection and identified by liquid chromatography electrospray ionization tandem mass spectrometry. Calibration curves were linear in the range of 25-2500 ppm for MUS and 40-2500 ppm for IBO, respectively. This method was successfully applied to identify and quantify MUS and IBO in Amanita mushrooms naturally grown and circulated in the drug market.

  18. Optimizing hydrophilic interaction liquid chromatography for ultrasensitive proteome analysis

    NARCIS (Netherlands)

    Di Palma, S.

    2013-01-01

    In the last decade, the field of proteomics has rapidly progressed with substantial advances in many aspects, particularly nano-liquid chromatographic (LC) separation, mass spectrometric (MS) instrumentation and bioinformatics tools. However, significant improvements are still needed to generate

  19. Analysis of drugs of abuse in human plasma by dispersive liquid-liquid microextraction and high-performance liquid chromatography.

    Science.gov (United States)

    Fernández, P; Regenjo, M; Bermejo, A M; Fernández, A M; Lorenzo, R A; Carro, A M

    2015-04-01

    Opioids and cocaine are widely used at present, both for recreational purposes and as drugs of abuse. This raises the need to develop new analytical methods specifically designed for the simultaneous detection of several drugs of abuse in biological samples. In this work, dispersive liquid-liquid microextraction (DLLME) was assessed as a new sample treatment for the simultaneous extraction of morphine (MOR), 6-acetylmorphine (6AM), cocaine (COC), benzoylecgonine (BZE) and methadone (MET) from human plasma. Preliminary assays were done before developing an experimental design based on a Uniform Network Doehlert which allowed the optimum extraction conditions to be identified, namely: a volume of extractant solvent (chloroform) and dispersant solvent (acetonitrile) of 220 µl and 3.2 ml, respectively; 0.2 g of NaCl as a salting-out additive; pH 10.6 and ultrasound stirring for 3.5 min. The resulting extracts were analyzed by high-performance liquid chromatography with photodiode array detection (HPLC-PDA), using an XBridge® RP18 column (250 × 4.6 mm i.d., 5 µm particle size). Calibration graphs were linear over the concentration range 0.1-10 µg ml⁻¹, and detection limits ranged from 13.9 to 28.5 ng ml⁻¹. Precision calculated at three different concentration levels in plasma was included in the range 0.1-6.8% RSD. Recoveries of the five drugs were all higher than 84% on average. Finally the proposed method was successfully applied to 22 plasma samples from heroin, cocaine and/or methadone users, and the most frequently detected drug was benzoylecgonine, followed by methadone, cocaine and morphine. Copyright © 2014 John Wiley & Sons, Ltd.

  20. Comprehensive Profiling of Phytohormones in Honey by Sequential Liquid-Liquid Extraction Coupled with Liquid Chromatography-Mass Spectrometry.

    Science.gov (United States)

    Wang, Qing; Cai, Wen-Jing; Yu, Lei; Ding, Jun; Feng, Yu-Qi

    2017-01-25

    Honey exhibits various nutritional and medicinal functions, which are highly related to the active components; thus, the exploration of new compounds in honey is of great importance. Because honey is a byproduct of flower nectar, which is rich in phytohormones, the existence of phytohormones in honey is anticipated. In this research, a method for comprehensive profiling of 49 phytohormones in honey was developed by sequential liquid-liquid extraction (LLE) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Good linearities for 49 phytohormones were obtained with correlation coefficients (R) larger than 0.9913. The limits of detection (LODs) were in the range of 0.2-628.2 pg/mL. Satisfied reproducibility and reliability were achieved by evaluation of the intra- and interday precisions with relative standard deviations (RSDs) less than 15.8% and relative recoveries ranging from 80.4 to 123.7%. The method was further applied to analyze the phytohormones in 14 monofloral raw honey samples and 3 commercial honey samples. The existence of 34 phytohormones was confirmed, including 14 cytokinins (CKs), 8 gibberellins (GAs), 5 brassinosteroids (BRs), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), abscisic acid (ABA), salicylic acid (SA), jasmonic acid (JA), jasmonoyl-leucine (JA-Leu), and jasmonoyl-phenylalanine (JA-Phe). In addition, the content and species of phytohormones varies in different kinds of honey. The study is beneficial to fully illustrate the phytohormone profile of honey and contributive to elucidate the mechanism of its nutritional and medicinal functions.

  1. Determination of tetraalkyllead compounds in gasoline by liquid chromatography-atomic absorption spectrometry

    Science.gov (United States)

    Messman, J.D.; Rains, T.C.

    1981-01-01

    A liquid chromatography-atomic absorption spectrometry (LC-AAS) hybrid analytical technique is presented for metal speciation measurements on complex liquid samples. The versatility and inherent metal selectivity of the technique are Illustrated by the rapid determination of five tetraalkyllead compounds in commercial gasoline. Separation of the individual tetraalkyllead species is achieved by reversed-phase liquid chromatography using an acetonitrile/water mobile phase. The effluent from the liquid Chromatograph Is introduced directly into the aspiration uptake capillary of the nebulizer of an air/acetylene flame atomic absorption spectrometer. Spectral interferences due to coeluting hydrocarbon matrix constituents were not observed at the 283.3-nm resonance line of lead used for analysis. Detection limits of this LC-AAS hydrid analytical technique, based on a 20-??L injection, are approximately 10 ng Pb for each tetraalkyllead compound.

  2. Micro scale self-interaction chromatography of proteins: A mAb case-study.

    Science.gov (United States)

    Hedberg, S H M; Heng, J Y Y; Williams, D R; Liddell, J M

    2016-02-19

    Self-interaction chromatography is known to be a fast, automated and promising experimental technique for determination of B22, but with the primary disadvantage of needing a significant amount of protein (>50 mg). This requirement compromises its usage as a technique for the early screening of new biotherapeutic candidates. A new scaled down SIC method has been evaluated here using a number of micro LC columns of different diameters and lengths, using typically 10 times less stationary phase than traditional SIC. Scale-down was successfully accomplished using these micro-columns, where the SIC results for a range of differing columns sizes were in agreement, as reflected by k', B22 and column volumes data. The results reported here demonstrate that a scaled down version of SIC can be easily implemented using conventional liquid chromatography system where the final amount of mAbs used was 10 times less than required by conventional SIC methodologies. Copyright © 2016. Published by Elsevier B.V.

  3. Lipidomic analysis of biological samples: Comparison of liquid chromatography, supercritical fluid chromatography and direct infusion mass spectrometry methods.

    Science.gov (United States)

    Lísa, Miroslav; Cífková, Eva; Khalikova, Maria; Ovčačíková, Magdaléna; Holčapek, Michal

    2017-11-24

    Lipidomic analysis of biological samples in a clinical research represents challenging task for analytical methods given by the large number of samples and their extreme complexity. In this work, we compare direct infusion (DI) and chromatography - mass spectrometry (MS) lipidomic approaches represented by three analytical methods in terms of comprehensiveness, sample throughput, and validation results for the lipidomic analysis of biological samples represented by tumor tissue, surrounding normal tissue, plasma, and erythrocytes of kidney cancer patients. Methods are compared in one laboratory using the identical analytical protocol to ensure comparable conditions. Ultrahigh-performance liquid chromatography/MS (UHPLC/MS) method in hydrophilic interaction liquid chromatography mode and DI-MS method are used for this comparison as the most widely used methods for the lipidomic analysis together with ultrahigh-performance supercritical fluid chromatography/MS (UHPSFC/MS) method showing promising results in metabolomics analyses. The nontargeted analysis of pooled samples is performed using all tested methods and 610 lipid species within 23 lipid classes are identified. DI method provides the most comprehensive results due to identification of some polar lipid classes, which are not identified by UHPLC and UHPSFC methods. On the other hand, UHPSFC method provides an excellent sensitivity for less polar lipid classes and the highest sample throughput within 10min method time. The sample consumption of DI method is 125 times higher than for other methods, while only 40μL of organic solvent is used for one sample analysis compared to 3.5mL and 4.9mL in case of UHPLC and UHPSFC methods, respectively. Methods are validated for the quantitative lipidomic analysis of plasma samples with one internal standard for each lipid class. Results show applicability of all tested methods for the lipidomic analysis of biological samples depending on the analysis requirements

  4. Application in pesticide analysis: Liquid chromatography - A review of the state of science for biomarker discovery and identification

    Science.gov (United States)

    Book Chapter 18, titled Application in pesticide analysis: Liquid chromatography - A review of the state of science for biomarker discovery and identification, will be published in the book titled High Performance Liquid Chromatography in Pesticide Residue Analysis (Part of the C...

  5. Vortex-assisted liquid-liquid-liquid microextraction (VALLLME) technique: A new microextraction approach for direct liquid chromatography and capillary electrophoresis analysis.

    Science.gov (United States)

    Makahleh, Ahmad; Yap, Hui Fang; Saad, Bahruddin

    2015-10-01

    A new, rapid and sensitive microextraction technique named vortex-assisted liquid-liquid-liquid microextraction (VALLLME) is proposed. The complete extraction process involves two steps. First, a vortex-assisted liquid-liquid microextraction (VALLME) procedure was used to extract the analytes from a relatively large volume of sample (donor phase) to a small volume of organic solvent (intermediate phase). Next, a micro-vortex-assisted liquid-liquid extraction (µ-VALLE) was used to extract the target analytes from the intermediate phase to a smaller volume of aqueous solution (acceptor phase). The final extract (acceptor phase) can be directly injected into the high performance liquid chromatography or capillary electrophoresis units without any further treatments. The selection of the intermediate phase and the manipulation of pH are key parameters that ensure good extraction efficiency of the technique. The proposed technique has been successfully applied for the determination of carvedilol (used as model analyte) in biological fluid samples. The optimum extraction conditions were: toluene as intermediate phase (150 μL); pH of the donor phase, 9.5; vortex time of the VALLME, 45 s (maximum speed, 2500 rpm); 0.1M HCl (15 μL) as acceptor phase; vortexing time of the µ-VALLME, 75 s (maximum stirring speed, 2500 rpm) and salt concentration in the donor phase, 5% (w/v). Under these conditions, enrichment factors of 51- and 418-fold for VALLME step and VALLLME procedure, respectively, were achieved. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Buffer-Free High Performance Liquid Chromatography Method for ...

    African Journals Online (AJOL)

    Purpose: To develop and validate a simple, economical and reproducible high performance liquid chromatographic (HPLC) method for the determination of theophylline in pharmaceutical dosage forms. Method: Caffeine was used as the internal standard and reversed phase C-18 column was used to elute the drug and ...

  7. The analysis of aqueous mixtures using liquid chromatography-electrospray mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, Steven [Iowa State Univ., Ames, IA (United States)

    1999-02-12

    The focus of this dissertation is the use of chromatographic methods coupled with electrospray mass spectrometry (ES-MS) for the determination of both organic and inorganic compounds in aqueous solutions. The combination of liquid chromatography (LC) methods and ES-MS offers one of the foremost methods for determining compounds in complex aqueous solutions. In this work, LC-ES-MS methods are devised using ion exclusion chromatography, reversed phase chromatography, and ion exchange chromatography, as well as capillary electrophoresis (CE). For an aqueous sample, these LC-ES-MS and CE-ES-MS techniques require no sample preparation or analyte derivatization, which makes it possible to observe a wide variety of analytes as they exist in solution. The majority of this work focuses on the use of LC-ES-MS for the determination of unknown products and intermediates formed during electrochemical incineration (ECI), an experimental waste remediation process. This report contains a general introduction to the project and the general conclusions. Four chapters have been removed for separate processing. Titles are: Chapter 2: Determination of small carboxylic acids by ion exclusion chromatography with electrospray mass spectrometry; Chapter 3: Electrochemical incineration of benzoquinone in aqueous media using a quaternary metal oxide electrode in the absence of a soluble supporting electrolyte; Chapter 4: The determination of electrochemical incineration products of 4-chlorophenol by liquid chromatography-electrospray mass spectrometry; and Chapter 5: Determination of small carboxylic acids by capillary electrophoresis with electrospray mass spectrometry.

  8. Comparison of gas chromatography/isotope ratio mass spectrometry and liquid chromatography/isotope ratio mass spectrometry for carbon stable-isotope analysis of carbohydrates

    NARCIS (Netherlands)

    Moerdijk-Poortvliet, T.C.W.; Schierbeek, H.; Houtekamer, M.; van Engeland, T.; Derrien, D.; Stal, L.J.; Boschker, H.T.S.

    2015-01-01

    We compared gas chromatography/isotope ratio mass spectrometry (GC/IRMS) and liquid chromatography/isotope ratio mass spectrometry (LC/IRMS) for the measurement of d13C values in carbohydrates. Contrary to GC/IRMS, no derivatisation is needed for LC/IRMS analysis of carbohydrates. Hence, although

  9. Comparison of gas chromatography/isotope ratio mass spectrometry and liquid chromatography/isotope ratio mass spectrometry for carbon stable-isotope analysis of carbohydrates

    NARCIS (Netherlands)

    Moerdijk-Poortvliet, Tanja C. W.; Schierbeek, Henk; Houtekamer, Marco; van Engeland, Tom; Derrien, Delphine; Stal, Lucas J.; Boschker, Henricus T. S.

    2015-01-01

    We compared gas chromatography/isotope ratio mass spectrometry (GC/IRMS) and liquid chromatography/isotope ratio mass spectrometry (LC/IRMS) for the measurement of δ(13)C values in carbohydrates. Contrary to GC/IRMS, no derivatisation is needed for LC/IRMS analysis of carbohydrates. Hence, although

  10. Comparison of gas chromatography/isotope ratio mass spectrometry and liquid chromatography/isotope ratio mass spectrometry for carbon stable-isotope analysis of carbohydrates

    NARCIS (Netherlands)

    Moerdijk-Poortvliet, T.C.W.; Schierbeek, H.; Houtekamer, M.; van Engeland, T.; Derrien, D.; Stal, L.J.; Boschker, H.T.S.

    2015-01-01

    Rationale: We compared gas chromatography/isotope ratio mass spectrometry (GC/IRMS) and liquid chromatography/isotope ratio mass spectrometry (LC/IRMS) for the measurement of δ13C values in carbohydrates. Contrary to GC/IRMS, no derivatisation is needed for LC/IRMS analysis of carbohydrates. Hence,

  11. Scale-up of counter-current chromatography: demonstration of predictable isocratic and quasi-continuous operating modes from the test tube to pilot/process scale.

    Science.gov (United States)

    Sutherland, Ian; Hewitson, Peter; Ignatova, Svetlana

    2009-12-11

    Predictable scale-up from test tube derived distribution ratios and analytical-scale sample loading optimisation is demonstrated using a model sample system of benzyl alcohol and p-cresol in a heptane:ethyl acetate:methanol:water phase system with the new 18 L Maxi counter-current chromatography centrifuge. The versatility of having a liquid stationary phase with its high loading capacity and flexible operating modes is demonstrated at two different scales by separating and concentrating target compounds using a mixture of caffeine, vanillin, naringenin and carvone using a quasi-continuous technique called intermittent counter-current extraction.

  12. Detection of Geothermal Phosphite Using High Performance Liquid Chromatography

    Science.gov (United States)

    Pech, Herbe; Henry, Amanda; Khachikian, Crist S.; Salmassi, Tina M.; Hanrahan, Grady; Foster, Krishna L.

    2009-01-01

    Little is known about the pre-biotic mechanisms that initiated the bioavailability of phosphorus, an element essential to life. A better understanding of phosphorus speciation in modern earth environments representative of early earth, may help to elucidate the origins of bioavailable phosphorus. This paper presents the first quantitative measurements of phosphite in a pristine geothermal pool representative of early earth. Phosphite and phosphate were initially identified and quantified in geothermal pool and stream samples at Hot Creek Gorge near Mammoth Lakes, California using suppressed conductivity ion chromatography. Results confirmed the presence of 0.06 ± 0.02 μM of phosphite and 0.05 ± 0.01 μM of phosphate in a geothermal pool. In the stream, phosphite concentrations were below detection limit (0.04 μM) and phosphate was measured at 1.06 ± 0.36 μM. The presence of phosphite in the geothermal pool was confirmed using both chemical oxidation and ion chromatography/mass spectrometry. PMID:19921877

  13. Isotope Fractionation in Methane Reactions Studied by Gas Chromatography and Liquid Scintillation

    DEFF Research Database (Denmark)

    Andersen, Bertel Lohmann; Bidoglio, G.; Leip, A.

    1997-01-01

    Determination of C-14-marked methane by gas chromatography and liquid scintillation counting is shown to be useful in studies of isotope effects. Data on the specific activity is used to separate the contributions of (CH4)-C-14, and (CH4)-C-12 to the gas-chromatographic peak area. As an application...

  14. Sorption of catechins under conditions of reverse-phase high-efficiency liquid chromatography

    Science.gov (United States)

    Shafigulin, R. V.; Egorova, K. V.; Bulanova, A. V.

    2010-08-01

    The physico-chemical principles of catechin sorption from various polar solvents onto silica gel modified with octadecyl groups were studied. Thermodynamic characteristics of the sorption were calculated, and the applicability of different models of retention was demonstrated for catechins under the conditions of reverse-phase high-efficiency liquid chromatography.

  15. Coupling nanoliter high-performance liquid chromatography to inductively coupled plasma mass spectrometry for arsenic speciation.

    Science.gov (United States)

    Cheng, Heyong; Shen, Lihuan; Liu, Jinhua; Xu, Zigang; Wang, Yuanchao

    2017-12-23

    Nanoliter high-performance liquid chromatography shows low consumption of solvents and samples, offering one of the best choices for arsenic speciation in precious samples in combination with inuctively coupled plasma mass spectrometry. A systematic investigation on coupling nanoliter high-performance liquid chromatography to inductively coupled plasma mass spectrometry from instrument design to injected sample volume and mobile phase was performed in this study. Nanoflow mobile phase was delivered by flow splitting using a conventional high-pressure pump with reuse of mobile phase waste. Dead volume was minimized to 60 nL for the sheathless interface based on the previously developed nanonebulizer. Capillary columns for nanoliter high-performance liquid chromatography were found to be sensitive to sample loading volume. An apparent difference was also found between the mobile phases for nanoliter and conventional high-performance liquid chromatography. Baseline separation of arsenite, arsenate, monomethylarsenic, and dimethylarsenic was achieved within 11 min on a 15 cm C18 capillary column and within 12 min on a 25 cm strong anion exchange column. Detection limits of 0.9-1.8 μg/L were obtained with precisions variable in the range of 1.6-4.2%. A good agreement between determined and certified values of a certified reference material of human urine (GBW 09115) validated its accuracy along with good recoveries (87-102%). © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. An Advanced, Interactive, High-Performance Liquid Chromatography Simulator and Instructor Resources

    Science.gov (United States)

    Boswell, Paul G.; Stoll, Dwight R.; Carr, Peter W.; Nagel, Megan L.; Vitha, Mark F.; Mabbott, Gary A.

    2013-01-01

    High-performance liquid chromatography (HPLC) simulation software has long been recognized as an effective educational tool, yet many of the existing HPLC simulators are either too expensive, outdated, or lack many important features necessary to make them widely useful for educational purposes. Here, a free, open-source HPLC simulator is…

  17. Determination of ribavirin in human serum using liquid chromatography tandem mass spectrometry

    NARCIS (Netherlands)

    van der Lijke, Henk; Alffenaar, Jan-Willem C.; Kok, Wim Th.; Greijdanus, Ben; Uges, Donald R. A.

    2012-01-01

    A method has been developed for the determination of ribavirin in human serum for therapeutic drug monitoring purposes, using liquid chromatography electrospray ionization mass spectrometry. Separation was obtained with a mobile phase gradient starting and ending in 100% aqueous conditions using a

  18. Quantification of levetiracetam in plasma of neonates by ultra performance liquid chromatography-tandem mass spectrometry

    NARCIS (Netherlands)

    Blonk, Maren I.; van der Nagel, Bart C.; Smit, Liesbeth S.; Mathot, Ron A. A.

    2010-01-01

    A sensitive and specific method using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the determination of levetiracetam (LEV) in plasma of neonates. A plasma aliquot of 50 microl was deproteinized by addition of 500 microl methanol which contained 5

  19. High-Performance Liquid Chromatography in the Undergraduate Chemical Engineering Laboratory

    Science.gov (United States)

    Frey, Douglas D.; Guo, Hui; Karnik, Nikhila

    2013-01-01

    This article describes the assembly of a simple, low-cost, high-performance liquid chromatography (HPLC) system and its use in the undergraduate chemical engineering laboratory course to perform simple experiments. By interpreting the results from these experiments students are able to gain significant experience in the general method of…

  20. Characterization of typical chemical background interferences in atmospheric pressure ionization liquid chromatography-mass spectrometry

    NARCIS (Netherlands)

    Guo, Xinghua; Bruins, Andries P.; Covey, Thomas R.

    2006-01-01

    The structures and origins of typical chemical background noise ions in positive atmospheric pressure ionization liquid chromatography/mass spectrometry (API LC/MS) are investigated and summarized in this study. This was done by classifying chemical background ions using precursor and product ion

  1. Determination for Synthesis and Content of Tetrahydropalmatine Based on High Performance Liquid Chromatography

    OpenAIRE

    Yuanming Zhang

    2014-01-01

    Tetrahydropalmatine is a kind of food additive with useful medicine value (dietary supplement), the tetrahydropalmatine synthetic process by using high performance liquid chromatography method was researched in the study, the experiments show that the dissolution rate of active ingredients in the tetrahydropalmatine water extract synthesized by this method has increased and the amount of active ingredient has greatly improved.

  2. Comparison of high performance liquid chromatography and enzymatic analysis of soluble carbohydrates in loblolly pine

    Science.gov (United States)

    Patricia L. Faulkner; Michele M. Schoeneberger; Kim H. Ludovici

    1993-01-01

    Foliar tissue was collected from a field study designed to test impacts of atmospheric pollutants on loblolIy pine (Pinus taeda L.) seedlings. Standard enzymatic (ENZ) and high performance liquid chromatography (HPLC) methods were used to analyze the tissue for soluble sugars. A comparison of the methods revealed no significant diffennces in accuracy...

  3. DETERMINATION OF CHLOROPHENOLS, NITROPHENOLS, AND METHYLPHENOLS IN GROUND-WATER SAMPLES USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Science.gov (United States)

    A high performance liquid chromatography (HPLC) method was developed to quantitatively determine phenolic compounds and their isomers in aqueous samples. The HPLC method can analyze a mixture of 15 contaminants in the same analytical run with an analysis time of 25 minutes. The...

  4. DETERMINATION OF CHLOROPHEONIS, NITROPHENOIS AND METHYLPHENOIS IN GROUND-WATER SAMPLES USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Science.gov (United States)

    A high performance liquid chromatography (HPLC) method was developed to quantitatively determine phenolic compounds and their isomers in aqueous samples. The HPLC method can analyze a mixture of 15 contaminants in the same analytical run with an analysis time of 25 minutes. The...

  5. A liquid chromatography-mass spectrometry-based metabolome database for tomato

    NARCIS (Netherlands)

    Moco, S.I.A.; Bino, R.J.; Vorst, O.F.J.; Verhoeven, H.A.; Groot, de J.C.W.; Beek, van T.A.; Vervoort, J.J.M.; Vos, de C.H.

    2006-01-01

    For the description of the metabolome of an organism, the development of common metabolite databases is of utmost importance. Here we present the Metabolome Tomato Database (MoTo DB), a metabolite database dedicated to liquid chromatography-mass spectrometry (LC-MS)- based metabolomics of tomato

  6. Analysis of Currently Available Analgesic Tablets by Modern Liquid Chromatography: An Undergraduate Laboratory Introduction to HPLC.

    Science.gov (United States)

    Kagel, R. A.; Farwell, S. O.

    1983-01-01

    Background information, procedures, and results, are provided for an undergraduate experiment in which analgesic tablets are analyzed using liquid chromatography. The experiment, an improved, modified version of the Waters Associates Inc. experiment, is simple to prepare, requiring little glassware and minimal sample manipulation by students. (JN)

  7. Extraction and Purification of Glucoraphanin by Preparative High-Performance Liquid Chromatography (HPLC)

    Science.gov (United States)

    Lee, Iris; Boyce, Mary C.

    2011-01-01

    A student activity that focuses on the isolation of glucoraphanin from broccoli using preparative high-performance liquid chromatography (HPLC) is presented here. Glucoraphanin is a glucosinolate, whose byproducts are known to possess anticancer properties. It is present naturally at high levels in broccoli and other "Brassica" vegetables. This…

  8. High-performance liquid chromatography determination of dapsone, monoacetyldapsone, and pyrimethamine in filter paper blood spots

    DEFF Research Database (Denmark)

    Rønn, A M; Lemnge, M M; Angelo, H R

    1995-01-01

    A high-performance liquid chromatography method for the simultaneous analysis of dapsone (DDS), the major metabolite of DDS, monoacetyldapsone (MADDS), and pyrimethamine (PYR) was modified for capillary blood samples obtained by finger prick and dried on filter paper. Limit of quantitation using...

  9. Determination of selenium in biological samples by gas-liquid chromatography with electron-capture detection.

    Science.gov (United States)

    Poole, C F; Evans, N J; Wibberley, D G

    1977-06-01

    Selenium can be determined quantitatively in biological samples after nitric acid-magnesium nitrate digestion and formation of 5-nitropiazselenole, by extraction into toluene for gas-liquid chromatography with electron-capture detection. The method is suitable for the determination of selenium in orchard leaves, bovine liver and human placenta, hair, blood and urine.

  10. Deformation and degradation of polymers in ultra-high-pressure liquid chromatography

    NARCIS (Netherlands)

    Uliyanchenko, E.; van der Wal, S.; Schoenmakers, P.J.

    2011-01-01

    Ultra-high-pressure liquid chromatography (UHPLC) using columns packed with sub-2 μm particles has great potential for separations of many types of complex samples, including polymers. However, the application of UHPLC for the analysis of polymers meets some fundamental obstacles. Small particles

  11. Benzoin Condensation: Monitoring a Chemical Reaction by High-Pressure Liquid Chromatography

    Science.gov (United States)

    Bhattacharya, Apurba; Purohit, Vikram C.; Bellar, Nicholas R.

    2004-01-01

    High-pressure liquid chromatography (HPLC) is the preferred method of separating a variety of materials in complex mixtures such as pharmaceuticals, polymers, soils, food products and biological fluids and is also considered to be a powerful analytical tool in both academia and industry. The use of HPLC analysis as a means of monitoring and…

  12. Nitrate and nitrite content in bottled beverages by ion-pair high-performance liquid chromatography.

    Science.gov (United States)

    Song, Yang; Deng, Gui-Fang; Xu, Xiang-Rong; Chen, Yong-Hong; Chen, Feng; Li, Hua-Bin

    2013-01-01

    Nitrate and nitrite levels in six types of beverages--total of 292 individual samples from 73 brands (four bottles each)--from Guangzhou city in China were evaluated by ion-pair high-performance liquid chromatography. All samples contained nitrate. Nitrate and nitrite ranges were 0.43-46.08 and beverages.

  13. Simulated Moving Bed Chromatography: Separation and Recovery of Sugars and Ionic Liquid from Biomass Hydrolysates

    Science.gov (United States)

    Caes, Benjamin R.; Van Oosbree, Thomas R.; Lu, Fachuang; Ralph, John; Maravelias, Christos T.

    2015-01-01

    Simulated moving bed chromatography, a continuous separation method, enables the nearly quantitative recovery of sugar products and ionic liquid solvent from chemical hydrolysates of biomass. The ensuing sugars support microbial growth, and the residual lignin from the process is intact. PMID:23939991

  14. Determination of organophosphorus acids by thermo-spray liquid chromatography-mass spectrometry

    NARCIS (Netherlands)

    Wils, E.R.J.; Hulst, A.G.

    1988-01-01

    The determination of thirteen organophosphorus acids, hydrolysis products of nerve agents and pesticides, by a combination of ion-pair liquid chromatography on a reversed-phase C18 column and thermospray mass spectrometry was investigated. Ammonium acetate and three tetraalkylammonium salts with

  15. Determination of Peroxide-Based Explosives Using Liquid Chromatography with On-Line Infrared Detection

    NARCIS (Netherlands)

    Schulte-Ladbeck, Rasmus; Edelmann, Andrea; Quintas, Guillermo; Lendl, Bernhard; Karst, U.

    2006-01-01

    A nondestructive analytical method for peroxide-based explosives determination in solid samples is described. Reversed-phase high-performance liquid chromatography in combination with on-line Fourier transform infrared (FT-IR) detection is used for the analysis of triacetonetriperoxide (TATP) and

  16. Determination of Bedaquiline in Human Serum Using Liquid Chromatography-Tandem Mass Spectrometry

    NARCIS (Netherlands)

    Alffenaar, Jan-Willem C.; Bolhuis, Mathieu; van Hateren, Kai; Sturkenboom, Marieke; Akkerman, Onno; de Lange, Wiel; Greijdanus, Ben; van der Werf, Tjip; Touw, Daan

    Bedaquiline, a diarylquinoline for the treatment of multidrug-resistant tuberculosis (TB), relies on exposure-dependent killing. As data on drug exposure in specific populations are scarce, pharmacokinetic studies may be of interest. No simple and robust validated liquid chromatography-tandem mass

  17. Quantification and validation of ertapenem using a liquid chromatography-tandem mass spectrometry method

    NARCIS (Netherlands)

    van Rijn, S.P.; Wessels, A.M.A.; Greijdanus, B.; Touw, D.J.; Alffenaar, J.W.C.

    Ertapenem, a carbapenem, relies on time-dependent killing. Therapeutic drug monitoring (TDM) should be considered, when ertapenem is used in specific populations, to achieve optimal bactericidal activity and optimize drug-dosing regimens. No validated liquid chromatography-tandem mass spectrometry

  18. [Isolation of toxic peptides from Amanita phalloides and their analysis using high-performance liquid chromatography].

    Science.gov (United States)

    Hampl, J

    1993-08-05

    The objective of the work is isolation of toxic peptides from Amanita phalloides--amatoxins (alfa-, beta-, gamma-amanitin) and phallotoxins (phalloidin, phallacidin, phallisin, phallisacin) by liquid chromatography on Sephadex LH-20 according to Yocum modification. Seven main toxins were isolated in centigram amounts. The purity of the toxins isolated was verified by the characteristics of their absorbance spectra, by thin layer chromatography (TLC) and high-performance liquid chromatography on reversed phase (RP-HPLC). The fraction of acid phallotoxins which appears homogenous in TLC and Sephadex LH-20 was separated into 5 substances (four of which are phallotoxins) by preparative RP-HPLC technique. The toxins isolated are sufficiently pure to be used as standards in HPLC.

  19. Development of predictive quantitative retention-activity relationship models of alkaloids by mixed micellar liquid chromatography.

    Science.gov (United States)

    Chen, Yu; Wu, Li-ping; Chen, Cong; Ye, Li-ming

    2010-02-01

    The mixed micellar liquid chromatography is a mode that uses mixed micellar system of Brij35/SDS (85 : 15) as a mobile phase under adequate experimental conditions, can simulate the resting membrane potential and the conformation of the long hydrophilic polyoxyethylene chains remains unchanged. In this article, the applications of biopartitioning micellar chromatography, using mixed micellar system to describe and estimate bioactivities of alkaloids, has been focused. The BMC(Brij35/SDS)-QRAR models of half-life time, volume of distribution, plasma clearance and area under concentration-time curve were obtained using Brij35-SDS retention data. The aim is to take a look at the capability of the mixed micellar liquid chromatography model to describe and/or estimate the bioactivity of alkaloids. (c) 2009 John Wiley & Sons, Ltd.

  20. Ionic liquid-based air-assisted liquid-liquid microextraction followed by high performance liquid chromatography for the determination of five fungicides in juice samples.

    Science.gov (United States)

    You, Xiangwei; Chen, Xiaochu; Liu, Fengmao; Hou, Fan; Li, Yiqiang

    2018-01-15

    A novel and simple ionic liquid-based air-assisted liquid-liquid microextraction technique combined with high performance liquid chromatography was developed to analyze five fungicides in juice samples. In this method, ionic liquid was used instead of a volatile organic solvent as the extraction solvent. The emulsion was formed by pulling in and pushing out the mixture of aqueous sample solution and extraction solvent repeatedly using a 10mL glass syringe. No organic dispersive solvent was required. Under the optimized conditions, the limits of detection (LODs) were 0.4-1.8μgL(-1) at a signal-to-noise ratio of 3. The limits of quantification (LOQs) set as the lowest spiking levels with acceptable recovery in juices were 10μgL(-1), except for fludioxonil whose LOQ was 20μgL(-1). The proposed method was applied to determine the target fungicides in juice samples, and acceptable recoveries ranging from 74.9% to 115.4% were achieved. Copyright © 2017. Published by Elsevier Ltd.

  1. Separation of carbohydrates using hydrophilic interaction liquid chromatography.

    Science.gov (United States)

    Fu, Qing; Liang, Tu; Li, Zhenyu; Xu, Xiaoyong; Ke, Yanxiong; Jin, Yu; Liang, Xinmiao

    2013-09-20

    A strategy was developed to rapidly evaluate chromatographic properties of hydrophilic interaction chromatography (HILIC) columns for separating carbohydrates. Seven HILIC columns (Silica, Diol, TSK Amide-80, XAmide, Click Maltose, Click β-CD, and Click TE-Cys columns) were evaluated by using three monosaccharide and seven disaccharides as probes. The influence of column temperature on the peak shape and tautomerization of carbohydrates, as well as column selectivity were investigated. The influence of surface charge property on the retention was also studied by using glucose, glucuronic acid, and glucosamine, which indicated that buffer salt concentration and pH value in mobile phase was necessary to control the ionic interactions between ionic carbohydrates and HILIC columns. According to evaluation results, the XAmide column was selected as an example to establish experimental schemes for separation of complex mixtures of oligosaccharide. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. Large-scale analysis of in Vivo phosphorylated membrane proteins by immobilized metal ion affinity chromatography and mass spectrometry

    DEFF Research Database (Denmark)

    Nühse, Thomas S; Stensballe, Allan; Jensen, Ole N

    2003-01-01

    Global analyses of protein phosphorylation require specific enrichment methods because of the typically low abundance of phosphoproteins. To date, immobilized metal ion affinity chromatography (IMAC) for phosphopeptides has shown great promise for large-scale studies, but has a reputation for poor...... specificity. We investigated the potential of IMAC in combination with capillary liquid chromatography coupled to tandem mass spectrometry for the identification of plasma membrane phosphoproteins of Arabidopsis. Without chemical modification of peptides, over 75% pure phosphopeptides were isolated from...... plasma membrane digests and detected and sequenced by mass spectrometry. We present a scheme for two-dimensional peptide separation using strong anion exchange chromatography prior to IMAC that both decreases the complexity of IMAC-purified phosphopeptides and yields a far greater coverage...

  3. The detection of nicotine in a Late Mayan period flask by gas chromatography and liquid chromatography mass spectrometry methods.

    Science.gov (United States)

    Zagorevski, Dmitri V; Loughmiller-Newman, Jennifer A

    2012-02-29

    Several ancient Mayan vessels from the Kislak Collection of the US Library of Congress were examined for the presence of alkaloids. One of them, a codex-style flask, bears a text that appears to read yo-'OTOT-ti 'u-MAY, spelling y-otoot 'u-may 'the home of its/his/her tobacco'. Samples extracted from this Late Classic period (600 to 900 AD) container were analyzed by gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS) methods. Nicotine was identified as the major component of the extracts. LC/MS analyses also yielded signals due to nicotine mono-oxides. The identities of the compounds were determined by comparison of the chromatographic and/or mass spectral characteristics with those from standards and literature data. High-resolution high mass accuracy tandem mass spectrometry (MS/MS) spectra of protonated nicotine and nicotine mono-oxides were measured to verify and to correct previous product ion assignments. These analyses provided positive evidence for nicotine from a Mayan vessel, indicating it as a likely holder of tobacco leafs. The result of this investigation is the first physical evidence of tobacco from a Mayan container, and only the second example where the vessel content recorded in a Mayan hieroglyphic text has been confirmed directly by chromatography/mass spectrometry trace analysis. Copyright © 2012 John Wiley & Sons, Ltd.

  4. Sensitive determination of nitrophenol isomers by reverse-phase high-performance liquid chromatography in conjunction with liquid-liquid extraction

    Science.gov (United States)

    A method for the highly sensitive determination of 2-, 3- and 4- nitrophenols was developed using reverse-phase high-performance liquid chromatography (RP-HPLC) with a UV photodiode array detector. Using a reverse-phase column and 40% aqueous acetonitrile as an eluent (i.e. isocratic elution), the i...

  5. Comparison of micellar electrokinetic capillary chromatography and high performance liquid chromatography on fingerprint of Cnidium monnieri.

    Science.gov (United States)

    Zhao, Luhua; Zhang, Xinyong; Tan, Xiying; Wu, Menghua; Xianga, Bingren

    2006-06-01

    In our studies, micellar electrokinetic capillary chromatography (MEKC) was employed in fingerprint analysis of Cnidium monnieri for the first time. Average chromatography of 10 batches Cnidium monnieri from Jiangsu province, China, which have long been considered as the original and genuine herbal medicine, was first established as the characteristic fingerprint. Within 25 min the major effective components were separated by 18 mM borate, 12 mM phosphate and 50 mM SDS (pH 9.2) containing 20% methanol. The relative standard deviations of migration times and peak areas were less than 5%. As a new approach of fingerprint, MKCE was compared to the conventional approach-HPLC in our experiments. The fingerprint developed by HPLC comprised 8 peaks that were collected within 40 min. Relative standard deviation (RSD) values of retention times of corresponding peaks in HPLC analysis were very small (maximum 3% and average 0.9%). In conclusion, each two methods had its advantages and disadvantages. Furthermore, besides HPLC, MEKC as a feasible method, could be used in the development of fingerprint of Cnidium monnieri.

  6. Comprehensive lipidomic analysis of human plasma using multidimensional liquid- and gas-phase separations: Two-dimensional liquid chromatography-mass spectrometry vs. liquid chromatography-trapped-ion-mobility-mass spectrometry.

    Science.gov (United States)

    Baglai, Anna; Gargano, Andrea F G; Jordens, Jan; Mengerink, Ynze; Honing, Maarten; van der Wal, Sjoerd; Schoenmakers, Peter J

    2017-12-29

    Recent advancements in separation science have resulted in the commercialization of multidimensional separation systems that provide higher peak capacities and, hence, enable a more-detailed characterization of complex mixtures. In particular, two powerful analytical tools are increasingly used by analytical scientists, namely online comprehensive two-dimensional liquid chromatography (LC×LC, having a second-dimension separation in the liquid phase) and liquid chromatography-ion mobility-spectrometry (LC-IMS, second dimension separation in the gas phase). The goal of the current study was a general assessment of the liquid-chromatography-trapped-ion-mobility-mass spectrometry (LC-TIMS-MS) and comprehensive two-dimensional liquid chromatography-mass spectrometry (LC×LC-MS) platforms for untargeted lipid mapping in human plasma. For the first time trapped-ion-mobility spectrometry (TIMS) was employed for the separation of the major lipid classes and ion-mobility-derived collision-cross-section values were determined for a number of lipid standards. The general effects of a number of influencing parameters have been inspected and possible directions for improvements are discussed. We aimed to provide a general indication and practical guidelines for the analyst to choose an efficient multidimensional separation platform according to the particular requirements of the application. Analysis time, orthogonality, peak capacity, and an indicative measure for the resolving power are discussed as main characteristics for multidimensional separation systems. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Determination of oxidized phosphatidylcholines by hydrophilic interaction liquid chromatography coupled to Fourier transform mass spectrometry.

    Science.gov (United States)

    Sala, Pia; Pötz, Sandra; Brunner, Martina; Trötzmüller, Martin; Fauland, Alexander; Triebl, Alexander; Hartler, Jürgen; Lankmayr, Ernst; Köfeler, Harald C

    2015-04-14

    A novel liquid chromatography-mass spectrometry (LC-MS) approach for analysis of oxidized phosphatidylcholines by an Orbitrap Fourier Transform mass spectrometer in positive electrospray ionization (ESI) coupled to hydrophilic interaction liquid chromatography (HILIC) was developed. This method depends on three selectivity criteria for separation and identification: retention time, exact mass at a resolution of 100,000 and collision induced dissociation (CID) fragment spectra in a linear ion trap. The process of chromatography development showed the best separation properties with a silica-based Kinetex column. This type of chromatography was able to separate all major lipid classes expected in mammalian samples, yielding increased sensitivity of oxidized phosphatidylcholines over reversed phase chromatography. Identification of molecular species was achieved by exact mass on intact molecular ions and CID tandem mass spectra containing characteristic fragments. Due to a lack of commercially available standards, method development was performed with copper induced oxidation products of palmitoyl-arachidonoyl-phosphatidylcholine, which resulted in a plethora of lipid species oxidized at the arachidonoyl moiety. Validation of the method was done with copper oxidized human low-density lipoprotein (LDL) prepared by ultracentrifugation. In these LDL samples we could identify 46 oxidized molecular phosphatidylcholine species out of 99 possible candidates.

  8. Capillary liquid chromatography combined with pressurized liquid extraction and dispersive liquid-liquid microextraction for the determination of vitamin E in cosmetic products.

    Science.gov (United States)

    Viñas, Pilar; Pastor-Belda, Marta; Campillo, Natalia; Bravo-Bravo, María; Hernández-Córdoba, Manuel

    2014-06-01

    Capillary liquid chromatography (LC) is used for the determination of tocopherols and tocotrienols in cosmetic products. Dispersive liquid-liquid microextraction (DLLME) allows the analytes to be preconcentrated into a very small volume of organic solvent which is then injected into the chromatograph running at a very low flow rate. Pressurized liquid extraction (PLE) at a high temperature and pressure was used to isolate vitamin E forms from cosmetics. The Taguchi experimental method was used to optimize the factors affecting DLLME. The parameters selected were 2mL of acetonitrile (disperser solvent), 100μL carbon tetrachloride (extraction solvent) and 10mL aqueous solution. A volume of 5μL of the organic phase was injected into the reversed-phase capillary LC system equipped with a diode array detector and using an isocratic mobile phase composed of an 95:5 (v/v) methanol:water mixture at a flow-rate of 20μLmin(-1). Quantification was carried out using aqueous standards and detection limits were in the range 0.1-0.5ngmL(-1), corresponding to 3-15ngg(-1) in the cosmetic sample. The recoveries were in the 87-105% range, with RSDs lower than 7.8%. The method was validated according to international guidelines and using a certified reference material. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Determination of insecticides in water using in situ halide exchange reaction-assisted ionic liquid dispersive liquid-liquid microextraction followed by high-performance liquid chromatography.

    Science.gov (United States)

    Li, Songqing; Gao, Haixiang; Zhang, Jiaheng; Li, Yubo; Peng, Bing; Zhou, Zhiqiang

    2011-11-01

    A dispersive liquid-liquid microextraction (DLLME) method using in situ halide exchange reaction to form ionic liquid (IL) extraction phase was developed to determine four insecticides (i.e. methoxyfenozide, tetrachlorvinphos, thiamethoxam, and diafenthiuron) in water samples. The preconcentration procedure, followed by high-performance liquid chromatography and variable wavelength detectors (VWD), enabled the formation of the immiscible IL extraction phase; the insecticides were transferred into the IL phase simultaneously, which enhanced the efficiency and sufficiency, greatly shortening the operation time. The experimental parameters affecting the extraction efficiency including volume of extraction IL, extraction and centrifugation times, volume of the sample solution and exchanging reagent, and addition of organic solvent and salt were investigated and optimized. Under optimized conditions, the extractions yielded recoveries of the target analytes from 82 to 102%. The calibration curves were linear, and the correlation coefficient ranged from 0.9990 to 0.9999 under the concentration levels of 5-200 μg/L. The relative standard deviation (n=6) was 2.9-4.6%. The limits of detection (LODs) for the four insecticides were between 0.98 and 2.54 μg/L. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Ionic-liquid-based dispersive liquid-liquid microextraction coupled with high-performance liquid chromatography for the forensic determination of methamphetamine in human urine.

    Science.gov (United States)

    Wang, Ruifeng; Qi, Xiujuan; Zhao, Lei; Liu, Shimin; Gao, Shuang; Ma, Xiangyuan; Deng, Youquan

    2016-07-01

    Determination of methamphetamine in forensic laboratories is a major issue due to its health and social harm. In this work, a simple, sensitive, and environmentally friendly method based on ionic liquid dispersive liquid-liquid microextraction combined with high-performance liquid chromatography was established for the analysis of methamphetamine in human urine. 1-Octyl-3-methylimidazolium hexafluorophosphate with the help of disperser solvent methanol was selected as the microextraction solvent in this process. Various parameters affecting the extraction efficiency of methamphetamine were investigated systemically, including extraction solvent and its volume, disperser solvent and its volume, sample pH, extraction temperature, and centrifugal time. Under the optimized conditions, a good linearity was obtained in the concentration range of 10-1000 ng/mL with determination coefficient >0.99. The limit of detection calculated at a signal-to-noise ratio of 3 was 1.7 ng/mL and the relative standard deviations for six replicate experiments at three different concentration levels of 100, 500, and 1000 ng/mL were 6.4, 4.5, and 4.7%, respectively. Meanwhile, up to 220-fold enrichment factor of methamphetamine and acceptable extraction recovery (>80.0%) could be achieved. Furthermore, this method has been successfully employed for the sensitive detection of a urine sample from a suspected drug abuser. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Bridging the gap between gas and liquid chromatography.

    Science.gov (United States)

    Gritti, Fabrice; Fogwill, Michael; Gilar, Martin; Jarrell, Joseph A

    2016-11-11

    The rapid and complete baseline separation of both volatile (C5 to C16 alkanes in gasoline or terpenes in plant extracts) and non-volatile (>C20 alkanes) organic compounds was achieved by combining (1) low-density fluid chromatography (LDFC) using carbon dioxide at elevated temperature (>90°C) and low pressure (1500psi) designed to increase the retention of the most volatile compounds and (2) high-vacuum technology (<10-4Torr) in order to preserve the maximum efficiency of short analytical columns (3.0mm×150mm packed with 1.8μm fully porous HSS-SB-C18 particles) when used in LDFC. The volatile compounds are eluted first under isobaric conditions (1500psi) in less than a minute followed by a linear gradient of the column back pressure (from 1500 to 3500psi in 5min) for the elution of the non-volatile compounds up to C40. The experimental results demonstrate that LDFC performed with short 3.0mm i.d. columns packed with sub-2μm particles and placed under adiabatic conditions enables the analysts to deliver a single, fast, and high-resolution separation of both volatile and non-volatile compounds. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Analysis of Camellia sinensis green and black teas via ultra high performance liquid chromatography assisted by liquid-liquid partition and two-dimensional liquid chromatography (size exclusion × reversed phase).

    Science.gov (United States)

    Scoparo, Camila T; de Souza, Lauro M; Dartora, Nessana; Sassaki, Guilherme L; Gorin, Philip A J; Iacomini, Marcello

    2012-01-27

    Green and black teas (Camellia sinensis) contain compounds ranging from simple phenolics to complex glycosides, many of which have well-recognized health benefits. Here, we describe two methodologies aiming to achieve a comprehensive analysis of hydro-alcoholic extracts of C. sinensis. In the first step, the extracts were partitioned in water, n-butanol, ethyl acetate and chloroform to separate the compounds according to their polarity, yielding less complex samples to be analyzed by ultra high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS). Additionally, a comprehensive two dimensional liquid chromatography (2D-LC) technique, employing size exclusion chromatography (SEC) × reversed phase (BEH-C18) was developed. The following compounds were identified on the basis of retention time, UV-spectra and MS fragmentation patterns: catechins, theaflavins and their gallate derivatives; kaempferol, quercetin and myricetin mono-, di-, tri- and tetraglycosides; esters of quinic acid and gallic or hydroxycinnamic acids; purine alkaloids, such as caffeine and theobromine and many lipids. Additionally, there were many novel compounds that were previously undescribed, such as saponin isomers and gallic acid esters of four glycosides of myricetin, quercetin and kaempferol. Copyright © 2011 Elsevier B.V. All rights reserved.

  13. [Determination of levamisole residue in animal livers by two liquid-liquid extraction steps-gas chromatography-mass spectrometry].

    Science.gov (United States)

    Xu, Jing; Xiao, Shanshan; Dong, Weifeng; Sui, Kai; Cao, Jijuan; Diao, Wenting; Zhang, Jing

    2012-09-01

    A novel method wasdeveloped for the determination oflevamisole in animal livers by liquid-liquid extraction and gas chromatography-mass spectrometry. Levamisole hydrochloride was transferred to levamisole in alkalinesolution, andthen extracted intoethyl acetate. Two liquid-liquid extraction steps were carried out with HCI aqueous solution and potassium hydroxide-dichloromethane system in turn, and the liposoluble substances and the water-soluble substances were removed, respectively. The qualitative and quantitative analyses of levamisole were achieved by gas chromatography-mass spectrometry (GC-MS) system. The characteristic fragments m/z 148, 176 and 204 were selected and m/z 204 was used as the quantitative ion. A good linear relationship was obtained between the peak area and the mass concentration of levamisole from 0.25 mg/L to 3.0 mg/L with the correlation coefficient of 0.999. The limit of quantitation (LOQ) for levamisole was 5 microg/kg, which was much lower than the international maximum residue limit. The average recoveries of levamisole spiked in chicken liver, duck liver, rabbit liver and pig liver samples at three spiked levels ranged from 76% to 106% with the relative standard deviations less than 9%. The method is simple and stable without tedious sample preparation, and suitable for the determination of levamisole in animal livers.

  14. [Determination of amygdalin in hawthorn by high performance liquid chromatography].

    Science.gov (United States)

    Lü, Weifeng; Ding, Mingyu

    2005-09-01

    A suitable method for extraction of amygdalin from hawthorn has been established. At first, the lipophilic components were removed with petroleum ether by ultrasonic extraction. The amygdalin was then extracted by methanol in a Soxhlet's apparatus. For quantitation, a high performance liquid chromatographic method was developed by using a reversed-phase C18 column, mobile phase of methanol-water (15:85, v/v) and a detection wavelengh of 215 nm. It can be concluded that the content of amygdalin is higher in the seeds than that in the hawthorn powder without the seeds and the yield of amygdalin is higher in the hawthorn pieces than that in the hawthorn powder.

  15. Quantitative analysis of multiple components based on liquid chromatography with mass spectrometry in full scan mode.

    Science.gov (United States)

    Xu, Min Li; Li, Bao Qiong; Wang, Xue; Chen, Jing; Zhai, Hong Lin

    2016-08-01

    Although liquid chromatography with mass spectrometry in full scan mode can obtain all the signals simultaneously in a large range and low cost, it is rarely used in quantitative analysis due to several problems such as chromatographic drifts and peak overlap. In this paper, we propose a Tchebichef moment method for the simultaneous quantitative analysis of three active compounds in Qingrejiedu oral liquid based on three-dimensional spectra in full scan mode of liquid chromatography with mass spectrometry. After the Tchebichef moments were calculated directly from the spectra, the quantitative linear models for three active compounds were established by stepwise regression. All the correlation coefficients were more than 0.9978. The limits of detection and limits of quantitation were less than 0.11 and 0.49 μg/mL, respectively. The intra- and interday precisions were less than 6.54 and 9.47%, while the recovery ranged from 102.56 to 112.15%. Owing to the advantages of multi-resolution and inherent invariance properties, Tchebichef moments could provide favorable results even in the situation of peaks shifting and overlapping, unknown interferences and noise signals, so it could be applied to the analysis of three-dimensional spectra in full scan mode of liquid chromatography with mass spectrometry. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. A novel urea-functionalized surface-confined octadecylimidazolium ionic liquid silica stationary phase for reversed-phase liquid chromatography.

    Science.gov (United States)

    Zhang, Mingliang; Tan, Ting; Li, Zhan; Gu, Tongnian; Chen, Jia; Qiu, Hongdeng

    2014-10-24

    One-pot synthesis of surface-confined ionic liquid functionalized silica spheres was proposed using N-(3-aminopropyl)imidazole, γ-isopropyltriethoxysilane and 1-bromooctadecane as starting materials. The surface modification of the silica spheres was successful with a high surface density of octadecylimidazolium, enabling the utilization of this new urea-functionalized ionic liquid-grafted silica material as stationary phase for high-performance liquid chromatography in reversed-phase mode. The long aliphatic chain combined with the multiple polar group embedded in the ligands imparted the new stationary phase fine selectivity towards PAH isomers and polar aromatics and higher affinity for phenolic compounds. The unique features of the new material, especially the effect of the urea group on the retention were elucidated by mathematic modeling. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Determination of diflubenzuron and chlorbenzuron in fruits by combining acetonitrile-based extraction with dispersive liquid-liquid microextraction followed by high-performance liquid chromatography.

    Science.gov (United States)

    Ruan, Chunqiang; Zhao, Xiang; Liu, Chenglan

    2015-09-01

    In this study, a simple and low-organic-solvent-consuming method combining an acetonitrile-partitioning extraction procedure followed by "quick, easy, cheap, effective, rugged and safe" cleanup with ionic-liquid-based dispersive liquid-liquid microextraction and high-performance liquid chromatography with diode array detection was developed for the determination of diflubenzuron and chlorbenzuron in grapes and pears. Ionic-liquid-based dispersive liquid-liquid microextraction was performed using the ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate as the extractive solvent and acetonitrile extract as the dispersive solvent. The main factors influencing the efficiency of the dispersive liquid-liquid microextraction were evaluated, including the extractive solvent type and volume and the dispersive solvent volume. The validation parameters indicated the suitability of the method for routine analyses of benzoylurea insecticides in a large number of samples. The relative recoveries at three spiked levels ranged between 98.6 and 109.3% with relative standard deviations of less than 5.2%. The limit of detection was 0.005 mg/kg for the two insecticides. The proposed method was successfully used for the rapid determination of diflubenzuron and chlorbenzuron residues in real fruit samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. An integrated process for the recovery of high added-value compounds from olive oil using solid support free liquid-liquid extraction and chromatography techniques.

    Science.gov (United States)

    Angelis, Apostolis; Hamzaoui, Mahmoud; Aligiannis, Nektarios; Nikou, Theodora; Michailidis, Dimitris; Gerolimatos, Panagiotis; Termentzi, Aikaterini; Hubert, Jane; Halabalaki, Maria; Renault, Jean-Hugues; Skaltsounis, Alexios-Léandros

    2017-03-31

    An integrated extraction and purification process for the direct recovery of high added value compounds from extra virgin olive oil (EVOO) is proposed by using solid support free liquid-liquid extraction and chromatography techniques. Two different extraction methods were developed on a laboratory-scale Centrifugal Partition Extractor (CPE): a sequential strategy consisting of several "extraction-recovery" cycles and a continuous strategy based on stationary phase co-current elution. In both cases, EVOO was used as mobile phase diluted in food grade n-hexane (feed mobile phase) and the required biphasic system was obtained by adding ethanol and water as polar solvents. For the sequential process, 17.5L of feed EVOO containing organic phase (i.e. 7L of EVOO treated) were extracted yielding 9.5g of total phenolic fraction corresponding to a productivity of 5.8g/h/L of CPE column. Regarding the second approach, the co-current process, 2L of the feed oil phase (containing to 0.8L of EVOO) were treated at 100mL/min yielding 1.03g of total phenolic fraction corresponding to a productivity of 8.9g/h/L of CPE column. The total phenolic fraction was then fractionated by using stepwise gradient elution Centrifugal Partition Chromatography (CPC). The biphasic solvent systems were composed of n-hexane, ethyl acetate, ethanol and water in different proportions (X/Y/2/3, v/v). In a single run of 4h on a column with a capacity of 1L, 910mg of oleocanthal, 882mg of oleacein, 104mg of hydroxytyrosol were successfully recovered from 5g of phenolic extract with purities of 85%, 92% and 90%, respectively. CPC fractions were then submitted to orthogonal chromatographic steps (adsorption on silica gel or size exclusion chromatography) leading to the isolation of additional eleven compounds belonging to triterpens, phenolic compounds and secoiridoids. Among them, elenolic acid ethylester was found to be new compound. Thin Layer Chromatography (TLC), Nuclear magnetic Resonance (NMR) and

  19. Determination of Niacinamide in Lotions and Creams Using Liquid-Liquid Extraction and High-Performance Liquid Chromatography

    Science.gov (United States)

    Usher, Karyn M.; Simmons, Carolyn R.; Keating, Daniel W.; Rossi, Henry F., III

    2015-01-01

    Chemical separations are an important part of an undergraduate chemistry curriculum. Sophomore students often get experience with liquid-liquid extraction in organic chemistry classes, but liquid-liquid extraction is not as often introduced as a quantitative sample preparation method in honors general chemistry or quantitative analysis classes.…

  20. Determination of Finasteride in Tablets by High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    K. Basavaiah

    2007-01-01

    Full Text Available A rapid, highly sensitive high performance liquid chromatographic method has been developed for the determination of finasteride(FNS in bulk drug and in tablets. FNS was eluted from a ODS C18 reversed phase column at laboratory temperature (30 ± 2°C with a mobile phase consisting of methanol and water (80+20 at a flow rate of 1 mL min-1 with UV detection at 225 nm. The retention time was ∼ 6.1 min and each analysis took not more than 10 min. Quantitation was achieved by measurement of peak area without using any internal standard. Calibration graph was linear from 2.0 to 30 μg mL-1 with limits of detection (LOD and quantification (LOQ being 0.2 and 0.6 μg mL-1, respectively. The method was validated according to the current ICH guidelines. Within-day co efficients of variation (CV ranged from 0.31 to 0.69% and between-day CV were in the range 1.2-3.2%. Recovery of FNS from the pharmaceutical dosage forms ranged from 97.89 – 102.9 with CV of 1.41-4.13%. The developed method was compared with the official method for FNS determination in its tablet forms.

  1. Liquid chromatography-mass spectrometric and liquid chromatography-tandem mass spectrometric determination of hallucinogenic indoles psilocin and psilocybin in "magic mushroom" samples.

    Science.gov (United States)

    Kamata, Tooru; Nishikawa, Mayumi; Katagi, Munehiro; Tsuchihashi, Hitoshi

    2005-03-01

    Accurate and sensitive analytical methods for psilocin (PC) and psilocybin (PB), tryptamine-type hallucinogens contained in "magic mushrooms," were investigated using liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS-MS). The chromatographic separation on an ODS column and mass spectral information gave complete discrimination between PC and PB without derivatization. The mass spectrometric detection had a high sensitivity, and the tandem mass spectrometric detection provided more specificity and accuracy, as well as high sensitivity. The detection limits ranged from 1 to 25 pg by LC-MS in the selected ion monitoring mode, and the intra- and inter-day coefficients of variation were estimated to be 4.21-5.93% by LC-MS-MS in the selected reaction monitoring mode. By applying the present LC-MS-MS technique to four real samples, the contents of PC and PB were found to vary over a wide range (0.60-1.4 and 0.18-3.8 mg/g dry wt. for PC and PB, respectively) between samples.

  2. Analyses of Indole Compounds in Sugar Cane (Saccharum officinarum L. Juice by High Performance Liquid Chromatography and Liquid Chromatography-Mass Spectrometry after Solid-Phase Extraction

    Directory of Open Access Journals (Sweden)

    Jean Wan Hong Yong

    2017-03-01

    Full Text Available Simultaneous quantitative analysis of 10 indole compounds, including indole-3-acetic acid (IAA, one of the most important naturally occurring auxins and some of its metabolites, by high performance liquid chromatography (HPLC and liquid chromatography-mass spectrometry (LC-MS after solid-phase extraction (SPE was reported for the first time. The analysis was carried out using a reverse phase HPLC gradient elution, with an aqueous mobile phase (containing 0.1% formic acid modified by methanol. Furthermore, a novel SPE procedure was developed for the pre-concentration and purification of indole compounds using C18 SPE cartridges. The combination of SPE, HPLC, and LC-MS was applied to screen for the indole compounds present in sugar cane (Saccharum officinarum L. juice, a refreshing beverage with various health benefits. Finally, four indole compounds were successfully detected and quantified in sugar cane juice by HPLC, which were further unequivocally confirmed by LC-MS/MS experiments operating in the multiple reaction monitoring (MRM mode.

  3. Monitoring of cefepime in human serum and plasma by micellar electrokinetic capillary chromatography: Improvement of sample preparation and validation by liquid chromatography coupled to mass spectrometry.

    Science.gov (United States)

    Šestáková, Nela; Theurillat, Regula; Sendi, Parham; Thormann, Wolfgang

    2017-04-01

    Cefepime monitoring in deproteinized human serum and plasma by micellar electrokinetic capillary chromatography and liquid chromatography coupled to mass spectrometry in presence of other drugs is reported. For micellar electrokinetic capillary chromatography, sample preparation comprised dodecylsulfate protein precipitation at pH 4.5 using an increased buffer concentration compared to that of a previous assay and removal of hydrophobic compounds with dichloromethane. This provided robust conditions for cefepime analysis in the presence of sulfamethoxazole and thus enabled its determination in samples of patients that receive cotrimoxazole. The liquid chromatography assay is based upon use of a column with a pentafluorophenyl-propyl modified and multiendcapped stationary phase and the coupling to electrospray ionization with a single quadrupole detector. The performances of both assays with multilevel internal calibration were assessed with calibration and control samples and both assays were determined to be robust. Cefepime levels monitored by micellar electrokinetic capillary chromatography in samples from patients that were treated with cefepime only and with cefepime and cotrimoxazole were found to compare well with those obtained by liquid chromatography coupled to mass spectrometry. Cefepime drug levels determined by micellar electrokinetic capillary chromatography could thereby be validated. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. High-throughput determination of cortisol, cortisone, and melatonin in oral fluid by on-line turbulent flow liquid chromatography interfaced with liquid chromatography/tandem mass spectrometry.

    Science.gov (United States)

    Fustinoni, Silvia; Polledri, Elisa; Mercadante, Rosa

    2013-07-15

    Cortisol, cortisone, and melatonin (CORTol, CORTone, and MELA, respectively) are hormones related to stress and sleep disorders. Their detection is relevant to epidemiological studies aimed at investigating the effects of circadian cycle disruption. The aim of this study was to develop and evaluate a high-throughput assay for the detection of CORTol, CORTone, and MELA concentrations in non-invasively collected oral fluid samples. A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method to measure levels of CORTol, CORTone, and MELA in oral fluid samples in the presence of deuterated analogs was optimized and validated. A 50 μL aliquot of oral fluid sample, obtained by centrifugation of a chewed swab, was purified using on-line turbulent flow liquid chromatography. Analytes were then separated using C18 reversed-phase chromatography, subjected to positive ionization using an electrospray source, then quantitated using a triple quadrupole mass detector in the selected reaction monitoring mode. Limits of quantification and linear dynamic ranges were found to be 0.55 nmol/L, 5.5 nmol/L, and 0.004 nmol/L, and up to 28 nmol/L, 277 nmol/L, and 0.43 nmol/L for CORTol, CORTone, and MELA, respectively. Inter- and intra-run precisions as relative standard deviation values were <5%, and accuracies were within 95-106% of theoretical concentrations. An evaluation of matrix effects showed that the use of deuterated analogs controlled sources of bias. Furthermore, the total analysis time per sample was 13 min, resulting in a throughput of approximately 100 samples/day. To our knowledge, this is the first automated, high-throughput assay for the simultaneous quantification of CORTol, CORTone, and MELA in oral fluid specimens. Copyright © 2013 John Wiley & Sons, Ltd.

  5. Online coupling of hydrophilic interaction/strong cation exchange/reversed-phase liquid chromatography with porous graphitic carbon liquid chromatography for simultaneous proteomics and N-glycomics analysis.

    Science.gov (United States)

    Zhao, Yun; Law, Henry C H; Zhang, Zaijun; Lam, Herman C; Quan, Quan; Li, Guohui; Chu, Ivan K

    2015-10-09

    In this study we developed a fully automated three-dimensional (3D) liquid chromatography methodology-comprising hydrophilic interaction separation as the first dimension, strong cation exchange fractionation as the second dimension, and low-pH reversed-phase (RP) separation as the third dimension-in conjunction downstream with additional complementary porous graphitic carbon separation, to capture non-retained hydrophilic analytes, for both shotgun proteomics and N-glycomics analyses. The performance of the 3D system alone was benchmarked through the analysis of the total lysate of Saccharomyces cerevisiae, leading to improved hydrophilic peptide coverage, from which we identified 19% and 24% more proteins and peptides, respectively, relative to those identified from a two-dimensional hydrophilic interaction liquid chromatography and low-pH RP chromatography (HILIC-RP) system over the same mass spectrometric acquisition time; consequently, the 3D platform also provided enhanced proteome and protein coverage. When we applied the integrated technology to analyses of the total lysate of primary cerebellar granule neurons, we characterized a total of 2201 proteins and 16,937 unique peptides for this primary cell line, providing one of its most comprehensive datasets. Our new integrated technology also exhibited excellent performance in the first N-glycomics analysis of cynomolgus monkey plasma; we successfully identified 122 proposed N-glycans and 135 N-glycosylation sites from 122 N-glycoproteins, and confirmed the presence of 38 N-glycolylneuraminic acid-containing N-glycans, a rare occurrence in human plasma, through tandem mass spectrometry for the first time. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Chiral separation and determination of ofloxacin enantiomers by ionic liquid-assisted ligand-exchange chromatography.

    Science.gov (United States)

    Bi, Wentao; Tian, Minglei; Row, Kyung Ho

    2011-01-21

    A simple and accurate method for the separation and determination of ofloxacin enantiomers was developed by ionic liquid-assisted ligand-exchange high performance liquid chromatography. Both achiral and chiral ionic liquids were tested for their efficiency of ofloxacin enantiomeric separation. The effects of ligands, concentration of metal ion, organic modifier, pH of the mobile phase, and temperature were also investigated and evaluated. Optimal conditions were obtained on a conventional C(18) column, where the mobile phase consisted of methanol/water (20 : 80, v/v) (containing 4.0 mmol L(-1) amino acid ionic liquid and 3.0 mmol L(-1) copper sulfate) at a flow rate of 0.5 mL min(-1). Under this condition, the ofloxacin enantiomers could be baseline separated within 14 minutes; the proposed method was used to analyze different commercial ofloxacin medicines.

  7. Hydrophilic interaction liquid chromatography: ESI-MS/MS of plasmalogen phospholipids from Pectinatus bacterium.

    Science.gov (United States)

    Rezanka, Tomáš; Siristova, Lucie; Matoulková, Dagmar; Sigler, Karel

    2011-08-01

    Liquid chromatography-electrospray tandem mass spectrometry (LC/ESI-MS/MS) was used to analyze phospholipids from three species of the anaerobic beer-spoilage bacterial genus Pectinatus. Analysis of total lipids by HILIC (Hydrophilic Interaction Liquid Chromatography) column succeeded in separating diacyl- and plasmalogen phospholipids. Plasmalogens were then analyzed by means of the ESI-MS/MS and more than 220 molecular species of four classes of plasmalogens (PlsCho (choline plasmalogen), PlsEtn (ethanolamine plasmalogen), PlsGro (glycerol plasmalogen), and PlsSer (serine plasmalogen)) were identified. Major molecular species were c-p19:0/15:0 PlsEtn and PlsSer, which accounted for more than 4% of the total lipids.

  8. High-pressure liquid chromatography with direct injection of gas sample.

    Science.gov (United States)

    Astanin, Anton I; Baram, Grigory I

    2017-06-09

    The conventional method of using liquid chromatography to determine the composition of a gaseous mixture entails dissolving vapors in a suitable solvent, then obtaining a chromatograph of the resulting solution. We studied the direct introduction of a gaseous sample into a C18 reversed-phase column, followed by separation of the components by HPLC with UV detection. Since the chromatography was performed at high pressure, vapors readily dissolved in the eluent and the substances separated in the column as effectively as in liquid samples. Samples were injected into the column in two ways: a) through the valve without a flow stop; b) after stopping the flow and relieving all pressure. We showed that an injectable gas volume could reach 70% of column dead volume. When an injected gaseous sample volume was less than 10% of the column dead volume, the resulting peaks were symmetrical and the column efficiency was high. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Arsenic speciation by liquid chromatography coupled with ionspray tandem mass spectrometry

    DEFF Research Database (Denmark)

    Corr, J. J.; Larsen, Erik Huusfeldt

    1996-01-01

    fragmentation patterns showing molecular dissociation through an expected common product ion were obtained for the four arsenosugars, Molecular mode detection was utilized for qualitative verification of speciation analysis by high-performance liquid chromatography coupled to inductively coupled plasma mass......Ionspray mass spectrometry, a well established organic analysis technique, has been coupled to high-performance liquid chromatography for speciation of organic arsenic compounds, The ionspray source and differentially pumped interface of the mass spectrometer were operated in dual modes...... for elemental and molecular analysis, Tandem mass spectrometry was employed to increase selectivity, Dual mode detection was applied to demonstrate the utility of the technique for analysis of nine organoarsenic standards, including four dimethylarsinylriboside derivatives (arsenosugars), Structural...

  10. Aerosol matrix-assisted laser desorption ionization for liquid chromatography/time-of-flight mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Murray, K.K.; Lewis, T.M.; Beeson, M.D.; Russell, D.H. (Texas A M Univ., College Station, TX (United States))

    1994-05-15

    We report the application of aerosol matrix-assisted laser desorption ionization (MALDI) to liquid chromatography/mass spectrometry (LC/MS). The aerosol MALDI experiment uses aerosol liquid introduction in conjunction with pulsed UV laser ionization to form ions from large biomolecules in solution. Mass analysis is achieved in a time-of-flight mass spectrometer. In the LC/MALDI-MS experiment, the matrix solution is combined with the column effluent in a mixing tee, LC/MALDI-MS is demonstrated for the separation of bradykinin, gramicidin S, and myoglobin. 32 refs., 8 figs., 1 tab.

  11. Determination of preservatives in cosmetics, cleaning agents and pharmaceuticals using fast liquid chromatography.

    Science.gov (United States)

    Baranowska, Irena; Wojciechowska, Iwona; Solarz, Natalia; Krutysza, Ewa

    2014-01-01

    This paper reports the development of a method for simultaneously determining five preservatives in cosmetics, cleaning agents and pharmaceuticals by fast liquid chromatography. Methylisothiazolinone, methylchloroisothiazolinone, benzyl alcohol, sodium benzoate and methylparaben were separated on a Chromolith Fast Gradient reversed-phase 18e column using gradient elution with acetonitrile and a 0.1% aqueous solution of formic acid, with a run time of 3 min. The preparation of solid and liquid samples included ultrasonic extraction with methanol with recoveries ranging from 69 to 119%. The developed method was used to analyze samples of cosmetics (66 samples), cleaning agents (five samples) and pharmaceutical industry products (17 samples).

  12. Phytochemical Profile of Erythrina variegata by Using High-Performance Liquid Chromatography and Gas Chromatography-Mass Spectroscopy Analyses

    Directory of Open Access Journals (Sweden)

    Suriyavathana Muthukrishnan

    2016-08-01

    Full Text Available Natural products derived from plant sources have been utilized to treat patients with numerous diseases. The phytochemical constituents present in ethanolic leaf extract of Erythrina variegata (ELEV were identified by using high-performance liquid chromatography (HPLC and gas chromatography-mass spectroscopy (GC-MS analyses. Shade dried leaves were powdered and extracted with ethanol for analyses through HPLC to identify selected flavonoids and through GC-MS to identify other molecules. The HPLC analysis of ELEV showed the presence of gallic and caffeic acids as the major components at concentrations of 2.0 ppm and 0.1 ppm, respectively, as well as other components. GC-MS analysis revealed the presence of 3-eicosyne; 3,7,11,15-tetramethyl-2-hexadecen-1-ol; butanoic acid, 3-methyl-3,7-dimethyl-6-octenyl ester; phytol; 1,2-benzenedicarboxylic acid, diundecyl ester; 1-octanol, 2-butyl-; squalene; and 2H-pyran, 2-(7-heptadecynyloxy tetrahydro-derivative. Because pharmacopuncture is a new evolving natural mode that uses herbal extracts for treating patients with various ailments with minimum pain and maximum effect, the results of this study are particularly important and show that ELEV possesses a wide range of phytochemical constituents, as indicated above, as effective active principle molecules that can be used individually or in combination to treat patients with various diseases.

  13. Phytochemical Profile of Erythrina variegata by Using High-Performance Liquid Chromatography and Gas Chromatography-Mass Spectroscopy Analyses.

    Science.gov (United States)

    Muthukrishnan, Suriyavathana; Palanisamy, Subha; Subramanian, Senthilkumar; Selvaraj, Sumathi; Mari, Kavitha Rani; Kuppulingam, Ramalingam

    2016-08-01

    Natural products derived from plant sources have been utilized to treat patients with numerous diseases. The phytochemical constituents present in ethanolic leaf extract of Erythrina variegata (ELEV) were identified by using high-performance liquid chromatography (HPLC) and gas chromatography-mass spectroscopy (GC-MS) analyses. Shade dried leaves were powdered and extracted with ethanol for analyses through HPLC to identify selected flavonoids and through GC-MS to identify other molecules. The HPLC analysis of ELEV showed the presence of gallic and caffeic acids as the major components at concentrations of 2.0 ppm and 0.1 ppm, respectively, as well as other components. GC-MS analysis revealed the presence of 3-eicosyne; 3,7,11,15-tetramethyl-2-hexadecen-1-ol; butanoic acid, 3-methyl-3,7-dimethyl-6-octenyl ester; phytol; 1,2-benzenedicarboxylic acid, diundecyl ester; 1-octanol, 2-butyl-; squalene; and 2H-pyran, 2-(7-heptadecynyloxy) tetrahydro-derivative. Because pharmacopuncture is a new evolving natural mode that uses herbal extracts for treating patients with various ailments with minimum pain and maximum effect, the results of this study are particularly important and show that ELEV possesses a wide range of phytochemical constituents, as indicated above, as effective active principle molecules that can be used individually or in combination to treat patients with various diseases. Copyright © 2016. Published by Elsevier B.V.

  14. Gas chromatography-mass spectrometry and high-performance liquid chromatography-diode array detection for dating of paper ink.

    Science.gov (United States)

    Díaz-Santana, Oscar; Vega-Moreno, Daura; Conde-Hardisson, Francisco

    2017-09-15

    An extraction and determination method is shown for the analysis of dyes and solvents present in two types of ballpoint pen inks that are deposited onto paper. Ink extracts are analysed using a combination of gas chromatography with mass spectrometry (GC-MS), and high-pressure liquid chromatography with photodiode array detection (HPLC-DAD), within a single sample extraction procedure. Seventeen solvents and thirteen dyes contained in two Montblanc® inks (black and blue) were monitored for 45 months at monthly intervals, in order to determine variations in the concentrations of the compounds over time. We also studied the relative variations between different compounds and the generation of degradation products such as phenol. The concentration data obtained from these compounds during their exposure have been analysed and a multiple regression model is developed for each ink type that allows an estimate of the exposure time of the ink on paper with a maximum error of between 4 and 7 months. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Polarity-based fractionation in proteomics: hydrophilic interaction vs reversed-phase liquid chromatography.

    Science.gov (United States)

    Jafari, M; Mirzaie, M; Khodabandeh, M; Rezadoost, H; Ghassempour, A; Aboul-Enein, H Y

    2016-07-01

    During recent decades, hydrophilic interaction liquid chromatography (HILIC) ahs been introduced to fractionate or purify especially polar solutes such as peptides and proteins while reversed-phase liquid chromatography (RPLC) is also a common strategy. RPLC is also a common dimension in multidimensional chromatography. In this study, the potential of HILIC vs RPLC chromatography was compared for proteome mapping of human peripheral blood mononuclear cell extract. In HILIC a silica-based stationary phase and for RPLC a C18 column were applied. Then separated proteins were eluted to an ion trap mass spectrometry system. Our results showed that the HILIC leads to more proteins being identified in comparison to RPLC. Among the total 181 identified proteins, 56 and 38 proteins were fractionated specifically by HILIC and RPLC, respectively. In order to demonstrate this, the physicochemical properties of identified proteins such as polarity and hydrophobicity were considered. This analysis indicated that polarity may play a major role in the HILIC separation of proteins vs RPLC. Using gene ontology enrichment analysis, it was also observed that differences in physicochemical properties conform to the cellular compartment and biological features. Finally, this study highlighted the potential of HILIC and the great orthogonality of RPLC in gel-free proteomic studies. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  16. Multiple scaling power in liquid gallium under pressure conditions

    Energy Technology Data Exchange (ETDEWEB)

    Li, Renfeng; Wang, Luhong; Li, Liangliang; Yu, Tony; Zhao, Haiyan; Chapman, Karena W.; Rivers, Mark L.; Chupas, Peter J.; Mao, Ho-kwang; Liu, Haozhe

    2017-06-01

    Generally, a single scaling exponent, Df, can characterize the fractal structures of metallic glasses according to the scaling power law. However, when the scaling power law is applied to liquid gallium upon compression, the results show multiple scaling exponents and the values are beyond 3 within the first four coordination spheres in real space, indicating that the power law fails to describe the fractal feature in liquid gallium. The increase in the first coordination number with pressure leads to the fact that first coordination spheres at different pressures are not similar to each other in a geometrical sense. This multiple scaling power behavior is confined within a correlation length of ξ ≈ 14–15 Å at applied pressure according to decay of G(r) in liquid gallium. Beyond this length the liquid gallium system could roughly be viewed as homogeneous, as indicated by the scaling exponent, Ds, which is close to 3 beyond the first four coordination spheres.

  17. Determination of food preservatives and saccharin by high-performance liquid chromatography.

    Science.gov (United States)

    Leuenberger, U; Gauch, R; Baumgartner, E

    1979-05-21

    The quantitative analysis of benzoic and sorbic acid, methyl, ethyl and propyl esters of p-hydroxybenzoic acid and saccharin in foodstuffs is described. These compounds are quantitatively extracted with disposable clean-up columns packed with Extrelut and simultaneously determined by high-performance liquid chromatography on reversed-phase columns. Complicated matrices such as cheese, cake, ketchup and chocolate were tested and recoveries were generally better than 95% in the concentration ranges normally used in the food industry.

  18. Measurement of teicoplanin by liquid chromatography-tandem mass spectrometry:development of a novel method

    OpenAIRE

    Fung, Francis H Y; Tang, Jonathan C Y; Hopkins, John P P; Dutton, John; Bailey, Lisa M; Davison, Andrew S

    2012-01-01

    Teicoplanin is an antibiotic used for the treatment of endocarditis, osteomyelitis, septic arthritis and methicillin-resistant Staphylococcus aureus. Teicoplanin is emerging as a suitable alternative antibiotic to vancomycin, where their trough serum levels are monitored by immunoassay routinely. This is the first report detailing the development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for measuring teicoplanin in patients' serum.

  19. Understanding and manipulating the separation in hydrophilic interaction liquid chromatography-a review

    OpenAIRE

    McCalley, D. V.

    2017-01-01

    Hydrophilic interaction liquid chromatography (HILIC) has emerged as a valuable complimentary technique to reversed-phase (RP), being especially suited for the analysis of polar and ionised solutes, which are difficult to retain in RP. For solutes amenable to both separation mechanisms, HILIC provides a different selectivity to RP, and also offers possibilities as an orthogonal mechanism for 2-dimensional LC when combined with RP. HILIC has further advantages of lower column back pressures, a...

  20. High-performance liquid chromatography for assaying NAD glycohydrolase from Neurospora crassa conidia.

    Science.gov (United States)

    Pietta, P; Pace, M; Menegus, F

    1983-06-01

    A rapid and sensitive high-performance liquid chromatographic technique was developed to determinate NAD glycohydrolase (EC 3.2.2.5.) activity from Neurospora crassa conidia. The separation of the assay substrate and products was achieved by isocratic reverse-phase chromatography and the peaks were detected by the absorbance at 259 nm. Quantities of NAD+ and nicotinamide as small as 10 pmol could be measured.

  1. Serum Protein Profile Study of Clinical Samples Using High Performance Liquid Chromatography-Laser Induced Fluorescence

    DEFF Research Database (Denmark)

    Karemore, Gopal Raghunath; Ukendt, Sujatha; Rai, Lavanya

    2009-01-01

    The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested...... for establishing the ability of HPLC-LIF protein profiling technique for discrimination, using hard clustering and Fuzzy clustering methods. The clustering algorithms have quite successfully classified the profiles as belonging to normal, cancer of cervix, and oral cancer conditions....

  2. The isolation and purification of glucoraphanin from broccoli seeds by solid phase extraction and preparative high performance liquid chromatography.

    Science.gov (United States)

    Rochfort, Simone; Caridi, Domenico; Stinton, Melanie; Trenerry, V Craige; Jones, Rod

    2006-07-07

    Plant foods contain not only essential nutrients, e.g. protein, amino acids, vitamins and minerals, but also phytochemicals that have added health benefits. One such class of phytochemicals are the glucosinolates. Glucosinolates, particularly glucoraphanin, are predominant in plants of the Brassica genus, most notably in vegetables such as broccoli. There is a growing interest in the role glucoraphanin plays in chemoprotection and as a result there is a requirement to accurately determine the levels of glucoraphanin in vegetable products. Reverse phase ion pair high performance liquid chromatography (HPLC) is the method of choice; however, this work has been hindered by the lack of available standard reference materials. Broccoli seeds, which are particularly rich in glucoraphanin (20-50 mg/g), have proved to be ideal for the isolation of glucoraphanin on the preparative scale. A novel preparative scale HPLC method with simple compound recovery has been developed to meet the need for a glucoraphanin standard.

  3. Intact-protein trapping columns for proteomic analysis in capillary high-performance liquid chromatography.

    Science.gov (United States)

    Guan, Xia; Yan, Guoquan; Gao, Mingxia; Hong, Guangfeng; Deng, Chunhui; Zhang, Xiangmin

    2010-10-29

    A new type of monolithic trapping columns with high mechanical strength was prepared by thin-layer sol-gel coating method and applied to trapping intact proteins for on-line capillary liquid chromatography. Monolithic trapping columns were fabricated by entrapping C8 reversed-phase particles into the capillary columns through a sol-gel network, which was formed by hydrolysis and polycondensation of methyltriethoxysilane. Hundreds times of trapping/untrapping for intact proteins were carried out. The trapping columns showed long-term stability up to 300 bar. Recovery, loading capacity and reproducibility of trapping columns were evaluated using four proteins. The recovery of four protein mixtures for the C8 monolithic trapping columns was 99.3% on average. The loading capacity of 5 mm × 320 μm i.d. C8 trapping columns for the protein mixtures was 30 μg. Day-to-day relative standard deviation (RSD) values for recoveries of protein mixtures on the same C8 trapping column ranged from 2.34 to 5.87%, column-to-column RSD values were from 3.01 to 6.81%. The C8 trapping columns were used to trap normal mouse liver intact proteins in a capillary liquid chromatography system. Results demonstrated high efficiency of the monolithic trapping columns for trapping intact proteins for proteomic analysis in on-line capillary liquid chromatography system. Copyright © 2010 Elsevier B.V. All rights reserved.

  4. Rapid determination of phthalate esters in alcoholic beverages by conventional ionic liquid dispersive liquid-liquid microextraction coupled with high performance liquid chromatography.

    Science.gov (United States)

    Fan, Yingying; Liu, Shuhui; Xie, Qilong

    2014-02-01

    A very simple, fast and environmentally friendly sample extraction method was proposed for the analysis of phthalate esters (PAEs, di-isobutyl phthalate (DIBP), dibutylphthalate (DBP), butylbenzylphthalate (BBP) and bis(2-ethylhexyl)phthalate (DEHP)) in alcoholic beverages by using conventional ionic liquid dispersive liquid-liquid microextraction. The samples were extracted by 160 μL 1-octyl-3-methylimidazolium hexafluorophosphate in the presence of appropriate amount of ethanol and 10% (w/v) sodium chloride solution; the enriched analytes in sedimented phases were analyzed by high performance liquid chromatography-diode array detector (HPLC-DAD). Under the optimum conditions, a satisfactory linearity (in the range of 0.02-1 μg mL(-1) for white spirits and 0.01-0.5 μg mL(-1) for red wines with the correlation coefficients (r) varying from 0.9983 to 1), acceptable recovery rates (88.5-103.5% for white spirits and 91.6-104.6% for red wines), good repeatability (RSD ≤ 8.0%) and low detection limits (3.1-4.2 ng mL(-1) for white spirits and 1.5-2.2 ng mL(-1) for red wines) were obtained. The developed method was successfully applied for the determination of the four PAEs in 30 white spirits and 11 red wines collected locally, and the DBP content in 63% (19:30) white spirits exceeded the specific migration limit of 0.3 mg kg(-1) established by international regulation. © 2013 Elsevier B.V. All rights reserved.

  5. Quantification of fat-soluble vitamins in human breast milk by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Kamao, Maya; Tsugawa, Naoko; Suhara, Yoshitomo; Wada, Akimori; Mori, Toshiyuki; Murata, Kazuo; Nishino, Riichiro; Ukita, Tetsuya; Uenishi, Kazuhiro; Tanaka, Kiyoshi; Okano, Toshio

    2007-11-15

    Sensitive quantification method for fat-soluble vitamins in human breast milk by liquid chromatography-tandem mass spectrometry was developed. Vitamins A, D and E were extracted from 10.0 mL of breast milk after saponifying by basic condition. Vitamin K derivatives were extracted from 3.0 mL of breast milk after lipase treatment. The corresponding stable isotope-labeled compounds were used as internal standards. For the determination of vitamin D compounds, derivatization with a Cookson-type reagent was performed. All fat-soluble vitamins were determined by liquid chromatography-tandem mass spectrometry in the positive ion mode. The detection limits of all analytes were 1-250 pg per 50 microL. The recoveries of fat-soluble vitamins were 91-105%. Inter-assay CV values of each vitamin were 1.9-11.9%. The mean concentrations of retinol, vitamin D3, 25-hydroxyvitamin D3, alpha-tocopherol, phylloquinone and menaquinone-4 were 0.455 microg/mL, 0.088 ng/mL, 0.081 ng/mL, 5.087 microg/mL, 3.771 ng/mL, and 1.795 ng/mL, respectively (n=82). This method makes possible to determine fat-soluble vitamins with a wide range of polarities in human breast milk. The assay may be useful for large-scale studies.

  6. Investigation of forced and total degradation products of amlodipine besylate by liquid chromatography and liquid chromatography-mass spectrometry

    Directory of Open Access Journals (Sweden)

    Stoiljković Zora Ž.

    2014-01-01

    Full Text Available An isocratic, reversed-phase liquid chromatographic method was applied for the investigation of the degradation products of amlodipine besylate under the stressed conditions in solution. Amlodipine besylate stock solutions were subjected to acid and alkali hydrolysis, chemical oxidation and photodegradation as well as to the electrochemical degradation by cyclic voltammetry in 0.05 mol/L NaHCO3 on gold electrode. The total degradation of amlodipine besylate was achieved in 5 mol/L NaOH at 80°C for 6 h and the compound with molecular formula C15H16NOCl was identified as a main degradation product. Under acidic (5 mol/L HCl at 80°C for 6 h stress conditions 75.2% of amlodipine besylate degradation was recorded. Oxidative degradation in the solution of 3% H2O2-methanol 80:20 at 80°C for 6 h showed that amlodipine besylate degraded to 80.1%. After 14 days of expose in photostability chamber amlodipine besylate solution showed degradation of 32.2%. In electrochemical degradation after 9 hours of cyclization the beginning of amlodipine oxidation was shifted for 200 mV to more negative potentials, with the degradation of 66.5%. Mass spectrometry analysis confirmed the presence of dehydro amlodipine derivate with molecular formula C20H23N2O5Cl in oxidative and acidic conditions while in electrochemical degradation was detected in traces. [Projekat Ministarsva nauke Republike Srbije, br. 172013

  7. Two-dimensional hydrophilic interaction/reversed-phase liquid chromatography for the preparative separation of polar and non-polar taxanes.

    Science.gov (United States)

    Rezadoost, Hassan; Ghassempour, Alireza

    2012-01-01

    Paclitaxel (Taxol®) is one of the best-known compounds of natural origin that has been used in the pharmaceutical industry. It is derived from taxanes extracted from different parts of yew trees (Taxaceae), cell cultures and taxane-producing endophytic fungi. Previous work reported a semi-preparative HPLC for the extraction of Paclitaxel (a non-polar taxane) from Taxus baccata L. and cell culture. In other work, 10-deacetylbaccatin III (a polar taxane) was separated from this medium at a preparative scale. To introduce a novel on-line two-dimensional heart-cut liquid chromatography using hydrophilic interaction/reversed-phase chromatography for preparative separation of polar and non-polar taxanes. To achieve this goal the system was developed from analytical to a larger preparative scale. In the first dimension of a semi-preparative column, taxanes were separated from polar compounds using a CN column, and in the second dimension the fraction containing taxanes was transferred to a C₁₈-column. Using this method polar and non-polar taxanes could be separated at analytical and semi-preparative scales. Combination of hydrophilic and reversed-phase chromatography in two-dimensional liquid chromatography (2D-LC) systems is a useful method for the separation of compounds with a large polarity range such as taxanes. Copyright © 2011 John Wiley & Sons, Ltd.

  8. Coupling of column liquid chromatography and surface-enhanced resonance Raman spectroscopy via a thin-layer chromatographic plate.

    NARCIS (Netherlands)

    Coulter, S.K.; Gooijer, C.; Velthorst, N.H.; Brinkman, U.A.T.; Somsen, G.W.

    1997-01-01

    Surface-enhanced resonance Raman (SERR) spectroscopy was used to characterize compounds separated by column liquid chromatography (LC). Three percent of the effluent from a conventional-size LC column were immobilized on a moving thinlayer chromatography (TLC) plate using a spray-jet

  9. Direct solid-phase microextraction combined with gas and liquid chromatography for the determination of lidocaine in human urine

    NARCIS (Netherlands)

    Koster, E.H M; Hofman, N.S K; de Jong, G.J.

    Solid-phase microextraction (SPME) has been combined with gas chromatography (GC) and liquid chromatography (LC) for the determination of lidocaine in human urine. A polydimethylsiloxane (PDMS) coated fibre was directly immersed into buffered urine. Extraction conditions such as time, pH, ionic

  10. Preparation and Characterization of a Polymeric Monolithic Column for Use in High-Performance Liquid Chromatography (HPLC)

    Science.gov (United States)

    Bindis, Michael P.; Bretz, Stacey Lowery; Danielson, Neil D.

    2011-01-01

    The high-performance liquid chromatography (HPLC) experiment, most often done in the undergraduate analytical instrumentation laboratory course, generally illustrates reversed-phase chromatography using a commercial C[subscript]18 silica column. To avoid the expense of periodic column replacement and introduce a choice of columns with different…

  11. Determination of polar organophosphorus pesticides in vegetables and fruits using liquid chromatography with tandem mass spectrometry: Selection of extraction solvent

    NARCIS (Netherlands)

    Mol, H.G.J.; Dam, R.C.J. van; Steijger, O.M.

    2003-01-01

    A method based on liquid chromatography (LC)-mass spectrometry (MS)/MS was developed for sensitive determination of a number of less gas chromatography (GC)-amenable organophosphorus pesticides (OPs; acephate, methamidophos, monocrotophos, omethoate, oxydemeton-methyl and vamidothion) in cabbage and

  12. Analysis of host-cell proteins in biotherapeutic proteins by comprehensive online two-dimensional liquid chromatography/mass spectrometry

    OpenAIRE

    Doneanu, Catalin E; Xenopoulos, Alex; Fadgen, Keith; Murphy, Jim; Skilton, St. John; Prentice, Holly; Stapels, Martha; Chen, Weibin

    2012-01-01

    Assays for identification and quantification of host-cell proteins (HCPs) in biotherapeutic proteins over 5 orders of magnitude in concentration are presented. The HCP assays consist of two types: HCP identification using comprehensive online two-dimensional liquid chromatography coupled with high resolution mass spectrometry (2D-LC/MS), followed by high-throughput HCP quantification by liquid chromatography, multiple reaction monitoring (LC-MRM). The former is described as a “discovery” assa...

  13. High-performance liquid chromatography analysis methods developed for quantifying enzymatic esterification of flavonoids in ionic liquids

    DEFF Research Database (Denmark)

    Lue, Bena-Marie; Guo, Zheng; Xu, X.B.

    2008-01-01

    Methods using reversed-phase high-performance liquid chromatography (RP-HPLC) with ELSD were investigated to quantify enzymatic reactions of flavonoids with fatty acids in the presence of diverse room temperature ionic liquids (RTILs). A buffered salt (preferably triethylamine-acetate) was found...... essential for separation of flavonoids from strongly polar RTILs, whereby RTILs were generally visible as two major peaks identified based on an ion-pairing/exchanging hypothesis. C8 and C12 stationary phases were optimal while mobile phase pH (3-7) had only a minor influence on separation. The method...... developed was successfully applied for primary screening of RTILs (> 20), with in depth evaluation of substrates in 10 RTILs, for their evaluation as reaction media....

  14. Quantitative analysis of benzodiazepines in vitreous humor by high-performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Elham Bazmi

    2016-08-01

    Full Text Available Objective: Benzodiazepines are frequently screened drugs in emergency toxicology, drugs of abuse testing, and in forensic cases. As the variations of benzodiazepines concentrations in biological samples during bleeding, postmortem changes, and redistribution could be biasing forensic medicine examinations, hence selecting a suitable sample and a validated accurate method is essential for the quantitative analysis of these main drug categories. The aim of this study was to develop a valid method for the determination of four benzodiazepines (flurazepam, lorazepam, alprazolam, and diazepam in vitreous humor using liquid–liquid extraction and high-performance liquid chromatography. Methods: Sample preparation was carried out using liquid–liquid extraction with n-hexane: ethyl acetate and subsequent detection by high-performance liquid chromatography method coupled to diode array detector. This method was applied to quantify benzodiazepines in 21 authentic vitreous humor samples. Linear curve for each drug was obtained within the range of 30–3000 ng/mL with coefficient of correlation higher than 0.99. Results: The limit of detection and quantitation were 30 and 100 ng/mL respectively for four drugs. The method showed an appropriate intra- and inter-day precision (coefficient of variation < 10%. Benzodiazepines recoveries were estimated to be over 80%. The method showed high selectivity; no additional peak due to interfering substances in samples was observed. Conclusion: The present method was selective, sensitive, accurate, and precise for the quantitative analysis of benzodiazepines in vitreous humor samples in forensic toxicology laboratory.

  15. Determination of parabens in serum by liquid chromatography-tandem mass spectrometry: Correlation with lipstick use.

    Science.gov (United States)

    Tahan, Gabriella Padovani; Santos, Nayara de Kássia Souza; Albuquerque, Ana Carolina; Martins, Isarita

    2016-08-01

    Parabens are the most widely used preservative and are considered to be relatively safe compounds. However, studies have demonstrated that they may have estrogenic activity, and there is ongoing debate regarding the safety and potential cancer risk of using products containing these compounds. In the present work, liquid chromatography-tandem mass spectrometry was applied to determine methylparaben and propylparaben concentrations in serum, and the results were correlated with lipstick application. Samples were analyzed using liquid-liquid extraction, followed by liquid chromatography-tandem mass spectrometry. The validation results demonstrated the linearity of the method over a range of 1-20 ng/mL, in addition to the method's precision and accuracy. A statistically significant difference was demonstrated between serum parabens in women who used lipstick containing these substances compared with those not using this cosmetic (p = 0.0005 and 0.0016, respectively), and a strong association was observed between serum parabens and lipstick use (Spearman correlation = 0.7202). Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Determination of prostaglandin analogs in cosmetic products by high performance liquid chromatography with tandem mass spectrometry.

    Science.gov (United States)

    Wittenberg, James B; Zhou, Wanlong; Wang, Perry G; Krynitsky, Alexander J

    2014-09-12

    A method was developed and validated for the determination of 16 prostaglandin analogs in cosmetic products. The QuEChERS (Quick, Easy, Cheap, Efficient, Rugged, Safe) liquid-liquid extraction method, typically used for pesticide residue analysis, was utilized as the sample preparation technique. The prostaglandin analogs were chromatographically separated and quantified using high performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). Thirty-one cosmetic products were surveyed, and 13 products were determined to contain a prostaglandin analog with amounts ranging from 27.4 to 297μg/g. The calculated concentrations for the cosmetic products were in a similar range when compared to the concentrations of three different prostaglandin analog-containing prescription products. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Determination of parabens using two microextraction methods coupled with capillary liquid chromatography-UV detection.

    Science.gov (United States)

    Chen, Chen-Wen; Hsu, Wen-Chan; Lu, Ya-Chen; Weng, Jing-Ru; Feng, Chia-Hsien

    2018-02-15

    Parabens are common preservatives and environmental hormones. As such, possible detrimental health effects could be amplified through their widespread use in foods, cosmetics, and pharmaceutical products. Thus, the determination of parabens in such products is of particular importance. This study explored vortex-assisted dispersive liquid-liquid microextraction techniques based on the solidification of a floating organic drop (VA-DLLME-SFO) and salt-assisted cloud point extraction (SA-CPE) for paraben extraction. Microanalysis was performed using a capillary liquid chromatography-ultraviolet detection system. These techniques were modified successfully to determine four parabens in 19 commercial products. The regression equations of these parabens exhibited good linearity (r2=0.998, 0.1-10μg/mL), good precision (RSDparabens from complex matrices. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. A novel high-throughput method for supported liquid extraction of retinol and alpha-tocopherol from human serum and simultaneous quantitation by liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Hinchliffe, Edward; Rudge, James; Reed, Paul

    2016-07-01

    Measurement of vitamin A (retinol) and E (alpha-tocopherol) in UK clinical laboratories is currently performed exclusively by high-performance liquid chromatography with ultraviolet detection. We investigated whether retinol and alpha-tocopherol could be measured simultaneously by liquid chromatography tandem mass spectrometry. Serum samples (100 μL) were extracted using Isolute + Supported Liquid Extraction plates. Chromatography was performed on a Phenomenex Kinetex Biphenyl 2.6 μm, 50 × 2.1 mm column, and liquid chromatography tandem mass spectrometry on a Waters Acquity TQD. Injection-to-injection time was 4.3 min. The assay was validated according to published guidelines. Patient samples were used to compare liquid chromatography tandem mass spectrometry and high-performance liquid chromatography with ultraviolet detection methods. For retinol and alpha-tocopherol, respectively, the assay was linear up to 6.0 and 80.0 μmol/L, and lower limit of quantification was 0.07 and 0.26 μmol/L. Intra and interassay imprecision were within desirable analytical specifications. Analysis of quality control material aligned to NIST SRM 968e, and relative spiked recovery from human serum, both yielded results within 15% of target values. Method comparison with high-performance liquid chromatography with ultraviolet detection methodology demonstrated a negative bias for retinol and alpha-tocopherol by the liquid chromatography tandem mass spectrometry method. Analysis of United Kingdom National External Quality Assurance Scheme samples yielded mean bias from the target value of +3.0% for retinol and -11.2% for alpha-tocopherol. We have developed a novel, high-throughput method for extraction of retinol and alpha-tocopherol from human serum followed by simultaneous quantitation by liquid chromatography tandem mass spectrometry. The method offers a rapid, sensitive, specific and cost-effective alternative to high-performance liquid chromatography with

  19. Determination of Alternaria mycotoxins in wine and juice using ionic liquid modified countercurrent chromatography as a pretreatment method followed by high-performance liquid chromatography.

    Science.gov (United States)

    Fan, Chen; Cao, Xueli; Liu, Man; Wang, Wei

    2016-03-04

    Alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TeA) are some of the main Alternaria mycotoxins that can be found as contaminants in food materials. The objective of this study was to develop a pretreatment method with countercurrent chromatography (CCC) for enrichment and cleanup of trace Alternaria mycotoxins in food samples prior to high-performance liquid chromatography (HPLC) analysis. An Analytical CCC instrument with a column volume 22.5mL was used, and a two-phase solvent system composed of ethyl acetate and water modified with 6% [HOOMIM][Cl] in mass to volume ratio was selected. Under the optimized CCC operation conditions, trace amounts of AOH, AME, and TeA in large volume of liquid sample were efficiently extracted and enriched in the stationary phase, and then eluted out just by reversing the stationary phase as mobile phase in the opposite flowing direction tail-to-head. The enrichment and elution strategies are unique and can be fulfilled online with high enrichment factors (87-114) and high recoveries (81.14-110.94%). The method has been successively applied to the determination of Alternaria mycotoxins in real apple juice and wine samples with the limits of detection (LOD) in the range of 0.03-0.14μgL(-1). Totally 12 wine samples and 15 apple juice samples from the local market were analyzed. The detection rate of AOH and AME in both kinds of the samples were more than 50%, while TeA was found in relatively high level of 1.75-49.61μgL(-1) in some of the apple juice samples. The proposed method is simple, rapid, and sensitive and could also be used for the analysis and monitoring of Alternaria mycotoxin in other food samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Concurrent triple-scale simulation of molecular liquids

    Science.gov (United States)

    Delgado-Buscalioni, Rafael; Kremer, Kurt; Praprotnik, Matej

    2008-03-01

    We present a triple-scale simulation of a molecular liquid, in which the atomistic, coarse-grained, and continuum descriptions of the liquid are concurrently coupled. The presented multiscale approach, which covers the length scales ranging from the micro- to macroscale, is a combination of two dual-scale models: a particle-based adaptive resolution scheme (AdResS), which couples the atomic and mesoscopic scales, and a hybrid continuum-molecular dynamics scheme (HybridMD). The combined AdResS-HybridMD scheme successfully sorts out the problem of large molecule insertion in the hybrid particle-continuum simulations of molecular liquids. The combined model is shown to correctly describe the hydrodynamics within a hybrid particle-continuum framework. The presented approach opens up the possibility to perform efficient grand-canonical molecular dynamics simulations of truly open molecular liquid systems.

  1. Ultrapreconcentration and determination of organophosphorus pesticides in water by solid-phase extraction combined with dispersive liquid-liquid microextraction and high-performance liquid chromatography.

    Science.gov (United States)

    Chen, Junhua; Zhou, Guangming; Deng, Yongli; Cheng, Hongmei; Shen, Jie; Gao, Yi; Peng, Guilong

    2016-01-01

    Solid-phase extraction coupled with dispersive liquid-liquid microextraction was developed as an ultra-preconcentration method for the determination of four organophosphorus pesticides (isocarbophos, parathion-methyl, triazophos and fenitrothion) in water samples. The analytes considered in this study were rapidly extracted and concentrated from large volumes of aqueous solutions (100 mL) by solid-phase extraction coupled with dispersive liquid-liquid microextraction and then analyzed using high performance liquid chromatography. Experimental variables including type and volume of elution solvent, volume and flow rate of sample solution, salt concentration, type and volume of extraction solvent and sample solution pH were investigated for the solid-phase extraction coupled with dispersive liquid-liquid microextraction with these analytes, and the best results were obtained using methanol as eluent and ethylene chloride as extraction solvent. Under the optimal conditions, an exhaustive extraction for four analytes (recoveries >86.9%) and high enrichment factors were attained. The limits of detection were between 0.021 and 0.15 μg/L. The relative standard deviations for 0.5 μg/L of the pesticides in water were in the range of 1.9-6.8% (n = 5). The proposed strategy offered the advantages of simple operation, high enrichment factor and sensitivity and was successfully applied to the determination of four organophosphorus pesticides in water samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Comprehensive two-dimensional liquid chromatography with on-line Fourier-transform-infrared-spectroscopy detection for the characterization of copolymers

    NARCIS (Netherlands)

    Kok, S.J.; Hankemeier, T.; Schoenmakers, P.J.

    2005-01-01

    The on-line coupling of comprehensive two-dimensional liquid chromatography (liquid chromatography × size-exclusion chromatography, LC × SEC) and infrared (IR) spectroscopy has been realized by means of an IR flow cell. The system has been assessed by the functional-group analysis of a series of

  3. Ultrahigh pressure liquid chromatography-(tandem) mass spectrometry in human sports drug testing: possibilities and limitations.

    Science.gov (United States)

    Thevis, Mario; Thomas, Andreas; Pop, Valentin; Schänzer, Wilhelm

    2013-05-31

    Doping control analytical laboratories for human sports predominantly employ nowadays chromatographic-mass spectrometric test methods for routine, high throughput screening and confirmation assays concerning low and high molecular mass analytes. Liquid chromatography-(tandem) mass spectrometry [(LC-MS(/MS)] and particularly ultrahigh pressure liquid chromatography (UHPLC)-MS/MS instruments have become devices of choice due to their indispensable capabilities that compensate for limitations inherent to other commonly used strategies such as immunological and gas chromatography-(tandem) mass spectrometry [(GC-MS(/MS)]-based detection methods. UHPLC-MS/MS-based assays at low mass spectrometric resolution have been established allowing for fast and sensitive targeted analyses focusing on pre-selected target analytes with diagnostic precursor-product ion pairs. Combining UHPLC to high resolution/high accuracy MS(/MS) further expanded the targeted approach (i.e., plotting extracted ion chromatograms of protonated or deprotonated molecules as well as product ions measured with accurate masses) toward non-targeted analyses enabling also retrospective data mining. In this review, recent applications of UHPLC-MS/MS in sports drug testing procedures published between 2008 and 2012 are presented and advantages as well as limitations in a short- and long-term perspective are discussed. Copyright © 2012 Elsevier B.V. All rights reserved.

  4. A simple, robust orthogonal background correction method for two-dimensional liquid chromatography.

    Science.gov (United States)

    Filgueira, Marcelo R; Castells, Cecilia B; Carr, Peter W

    2012-08-07

    Background correction is a very important step that must be performed before peak detection or any quantification procedure. When successful, this step greatly simplifies such procedures and enhances the accuracy of quantification. In the past, much effort has been invested to correct drifting baseline in one-dimensional chromatography. In fast online comprehensive two-dimensional liquid chromatography (LC×LC) coupled with a diode array detector (DAD), the change in the refractive index (RI) of the mobile phase in very fast gradients causes extremely serious baseline disturbances. The method reported here is based on the use of various existing baseline correction methods of one-dimensional (1D) liquid chromatography to correct the two-dimensional (2D) background in LC×LC. When such methods are applied orthogonally to the second dimension ((2)D), background correction is dramatically improved. The method gives an almost zero mean background level and it provides better background correction than does simple subtraction of a blank. Indeed, the method proposed does not require running a blank sample.

  5. Liquid chromatography/mass spectrometry and liquid chromatography/nuclear magnetic resonance as complementary analytical techniques for unambiguous identification of polymethoxylated flavones in residues from molecular distillation of orange peel oils (Citrus sinensis).

    Science.gov (United States)

    Weber, Berthold; Hartmann, Beate; Stöckigt, Detlef; Schreiber, Klaus; Roloff, Michael; Bertram, Heinz-Jürgen; Schmidt, Claus O

    2006-01-25

    Liquid chromatography/mass spectrometry and liquid chromatography/nuclear magnetic resonance techniques with ultraviolet/diode array detection were used as complementary analytical tools for the reliable identification of polymethoxylated flavones in residues from molecular distillation of cold-pressed peel oils of Citrus sinensis. After development of a liquid chromatographic separation procedure, the presence of several polymethoxy flavones such as sinensetin, nobiletin, tangeretin, quercetogetin, heptamethoxyflavone, and other derivatives was unambiguously confirmed. In addition, proceranone, an acetylated tetranortriterpenoid with limonoid structure, was identified for the first time in citrus.

  6. Preparative Isolation and Purification of Flavone C-Glycosides from the Leaves of Ficus microcarpa L. f by Medium-Pressure Liquid Chromatography, High-Speed Countercurrent Chromatography, and Preparative Liquid Chromatography.

    Science.gov (United States)

    Wang, Xiaohong; Liang, Yong; Zhu, Licai; Xie, Huichun; Li, Hang; He, Junting; Pan, Man; Zhang, Tianyou; Ito, Yoichiro

    2010-01-01

    Combined with medium-pressure liquid chromatography (MPLC) and preparative high-performance liquid chromatography (perp-HPLC), high-speed countercurrent chromatography (HSCCC) was applied for separation and purification of flavone C-glycosides from the crude extract of leaves of Ficus microcarpae L. f. HSCCC separation was performed on a two-phase solvent system composed of methyl tert- butyl ether - ethyl acetate - 1-butanol - acetonitrile - 0.1% aqueous trifluoroacetic acid at a volume ratio of 1:3:1:1:5. Partially resolved peak fractions from HSCCC separation were further purified by preparative HPLC. Four well-separated compounds were obtained and their purities were determined by HPLC. The purities of these peaks were 97.28%, 97.20%, 92.23%, and 98.40%.. These peaks were characterized by ESI-MS(n). According to the reference, they were identified as orientin (peak I), isovitexin-3″-O-glucopyranoside (peak II), isovitexin (peak III), and vitexin (peak IV), yielded 1.2 mg, 4.5 mg, 3.3 mg, and 1.8 mg, respectively.

  7. Performance and limits of liquid chromatography isotope ratio mass spectrometry system for halogenated compounds

    Science.gov (United States)

    Gilevska, Tetyana; Gehre, Matthias; Richnow, Hans

    2014-05-01

    Compound Specific Isotope Analysis (CSIA) has been an important step for the assessment of the origin and fate of compounds in environmental science.[1] Biologically or pharmaceutically important compounds often are not amenable for gas chromatographic separation because of high polarity and lacking volatility, thermostability. In 2004 liquid chromatography isotope ratio mass spectrometry (LC-IRMS) became commercially available. LC-IRMS system intent a quantitative conversion of analytes separation into CO2 via wet oxidation with sodium persulfate in the presence of phosphoric acid while analytes are still dissolved in the aqueous liquid phase.[2] The aim of this study is to analyze the oxidation capacity of the interface of the LC-IRMS system and determine which parameters could improve oxidation of compounds which are resistant to persulfate oxidation. Oxidation capacity of the liquid chromatography isotope ratio mass spectrometry system was tested with halogenated acetic acid and a set of aromatic compounds with different substitutes. Acetic acid (AA) was taken as a model compound for complete oxidation and compared to the oxidation of other analytes on a molar basis. Correct values were obtained for di- and mono chlorinated and fluorinated and also for tribrominated acetic acid and for all studied aromatic compounds. Incomplete oxidation for trichloroacetic (TCAA) and trifluoroacetic (TFAA) acid was revealed with lower recovery compared to acetic acid and isotope fractionation leading to depleted carbon isotope composition compared to values obtained with an elementary analyzer connected to an isotope mass spectrometer Several optimization steps were tried in order to improve the oxidation of TCAA and TFAA: (i) increasing the concentration of the oxidizing agent, (ii) variation of flow rate of the oxidizing and acid solution, (iii) variation of flow rate of liquid chromatography pump (iv) addition of a catalyzer. These modifications lead to longer reaction time

  8. Study of the separation limits of continuous solid support free liquid-liquid chromatography: separation of capsaicin and dihydrocapsaicin by centrifugal partition chromatography.

    Science.gov (United States)

    Goll, Johannes; Frey, Andreas; Minceva, Mirjana

    2013-04-05

    Sequential centrifugal partition chromatography (sCPC) is a cyclic solid support-free liquid-liquid chromatographic process, in which a continuously introduced feed mixture is separated into two sequentially collected product streams. The few experimental demonstrations of this concept already revealed its potential for the preparative separation of pharmaceuticals and fine chemicals. In this work not only the possibilities, but also the limits of the sCPC technology are explored. A feed mixture consisting of capsaicin and dihydrocapsaicin, whose molecular structure differs in only one double bond, was selected for this purpose. The sCPC unit operating parameters needed for a complete separation of the feed mixture were selected using the recently published approach, which uses the partition coefficient of the feed components and the hydrodynamic characteristics of the system as input data. A complete separation of capsaicin and dihydrocapsaicin with the solvent system heptane/ethyl acetate/methanol/water:1/1/1/1 (v/v/v/v) was achieved, although the separation factor was only 1.32. The sCPC unit separation performance was successfully simulated using the cell model. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. Low density solvent-based dispersive liquid-liquid microextraction for the determination of synthetic antioxidants in beverages by high-performance liquid chromatography.

    Science.gov (United States)

    Çabuk, Hasan; Köktürk, Mustafa

    2013-01-01

    A simple and efficient method was established for the determination of synthetic antioxidants in beverages by using dispersive liquid-liquid microextraction combined with high-performance liquid chromatography with ultraviolet detection. Butylated hydroxy toluene, butylated hydroxy anisole, and tert-butylhydroquinone were the antioxidants evaluated. Experimental parameters including extraction solvent, dispersive solvent, pH of sample solution, salt concentration, and extraction time were optimized. Under optimal conditions, the extraction recoveries ranged from 53 to 96%. Good linearity was observed by the square of correlation coefficients ranging from 0.9975 to 0.9997. The relative standard deviations ranged from 1.0 to 5.2% for all of the analytes. Limits of detection ranged from 0.85 to 2.73 ng mL(-1). The method was successfully applied for determination of synthetic antioxidants in undiluted beverage samples with satisfactory recoveries.

  10. Analysis of potential migrants from plastic materials in milk by liquid chromatography-mass spectrometry with liquid-liquid extraction and low-temperature purification.

    Science.gov (United States)

    Bodai, Zsolt; Szabó, Bálint Sámuel; Novák, Márton; Hámori, Susanne; Nyiri, Zoltán; Rikker, Tamás; Eke, Zsuzsanna

    2014-10-15

    A simple and fast analytical method was developed for the determination of six UV stabilizers (Cyasorb UV-1164, Tinuvin P, Tinuvin 234, Tinuvin 326, Tinuvin 327, and Tinuvin 1577) and five antioxidants (Irgafos 168, Irganox 1010, Irganox 3114, Irganox 3790, and Irganox 565) in milk. For sample preparation liquid-liquid extraction with low-temperature purification combined with centrifugation was used to remove fats, proteins, and sugars. After the cleanup step, the sample was analyzed with high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). External standard and matrix calibrations were tested. External calibration proved to be acceptable for Tinuvin P, Tinuvin 234, Tinuvin 326, Tinuvin 327, Irganox 3114, and Irganox 3790. The method was successfully validated with matrix calibration for all compounds. Method detection limits were between 0.25 and 10 μg/kg. Accuracies ranged from 93 to 109%, and intraday precisions were <13%.

  11. Liquid chromatography tandem-mass spectrometry (LC-MS/MS)--technique and applications in endocrinology.

    Science.gov (United States)

    Vogeser, M; Parhofer, K G

    2007-10-01

    In recent years, liquid chromatography tandem mass spectrometry (LC-MS/MS) has emerged as an innovative analytical technology applicable to a wide number of analyses in the endocrinology laboratory. Compared to the "traditional" technique of gas chromatography-mass spectrometry (GC-MS), LC-MS/MS is easier to use and is applicable for a substantially larger number of relevant analytes. With the development of LC-MS/MS, the widespread application of the proven principle of isotope dilution mass spectrometry is now feasible not only in research but also for routine applications. The aim of this review is to explain the basic technical principles of LC-MS/MS, to describe the general characteristics of analytical LC-MS/MS applications and to comprehensively discuss the application of this technology in the field of endocrinology.

  12. High-performance liquid chromatography of bile pigments: separation and characterization of the urobilinoids.

    Science.gov (United States)

    Bull, R V; Lim, C K; Gray, C H

    1981-11-20

    The detailed analysis of faecal bile pigments by high-performance liquid chromatography is described. Non-aqueous reversed-phase systems with acetonitrile-dimethyl sulphoxide or acetonitrile-dimethyl sulphoxide-methanol as the mobile phase on C1, C8 or C18-bonded silica are used for the group separation of verdinoids, violinoids and urobilinoids. A silica column, with acetonitrile-water-tetraethylene-pentamine as mobile phase, separates the laevorotatory stercobilin (C33H46N4O6) and half-stercobilin (C33H44N4O6) from the optically inactive urobilin (C33H42N4O6). The diastereoisomers are resolved by converting the urobilinoids into their dimethyl esters before chromatography on a silica column with n-heptane-methyl acetate-methanol containing 1% of diethylamine as the solvent system.

  13. Determination of 1-hydroxypyrene in human urine by high-performance liquid chromatography

    DEFF Research Database (Denmark)

    Hansen, Åse Marie; Poulsen, O M; Christensen, J M

    1993-01-01

    A high-performance liquid chromatography (HPLC)/fluorescence method for quantitative analysis of 1-hydroxypyrene in urine was developed. The method validation analysis showed the method to be in analytical control. No significant systematical errors could be demonstrated. The entire run time....... The developed method is presently used for measurement of 1-hydroxypyrene in urine samples from workers exposed to a low airborne level of polycyclic aromatic hydrocarbons, generally less than 25 micrograms/m3. The urine samples of exposed workers (n = 122) showed a range of 1-hydroxypyrene from the limit...... of chromatography was 10 min using isocratic elution (acetonitrile-water, 70:30), and the retention time for 1-hydroxypyrene was 3.5 min. The short run time in combination with the low limit detection (1.37 nmol/L) makes the method potentially applicable for surveillance of pyrene exposure in work environments...

  14. A method for fast determination of psoralens in oral solutions of phytomedicines using liquid chromatography.

    Science.gov (United States)

    Pires, Adriana Elias; Honda, Neli Kiko; Cardoso, Cláudia Andréa Lima

    2004-10-29

    A method for sample preparation and analysis by high performance liquid chromatography with UV detection (HPLC-UV) has been developed for routine analysis of psoralen and bergapten, photosensitizing compounds, in oral solutions of phytomedicines employed in Brazil for some illnesses. The linearity, accuracy, the inter- and intra-day precision of the procedure were evaluated. Calibration curves for psoralen and bergapten were linear in the range of 1.0-600.0 microg ml(-1) and 1.0-400.0 microg ml(-1) respectively. The recoveries of the psoralens in the oral solutions analysed were 94.43-99.97%. The percentage coefficient of variation (CV) of the quantitative analysis of the psoralens in the products analysis was within 5%. In inter-equipment study was employed gas chromatography-flame ionization (CG-FID) detection.

  15. Ion Exchange Chromatography-Indirect Ultraviolet Detection for Separation and Determination of Morpholinium Ionic Liquid Cations.

    Science.gov (United States)

    Zhang, Yu; Ma, Yajie; Yu, Hong; Liu, Yongqiang

    2017-01-01

    A rapid analytical method of ion exchange chromatography with indirect ultraviolet detection was developed to determine morpholinium ionic liquid (IL) cations, i.e. N-methyl-N-ethyl morpholinium cation ([MEMo]+) and N-methyl-N-propyl morpholinium cation ([MPMo]+). Chromatographic separation of morpholinium cations was performed on a sulfonic acid base cation exchange column using imidazolium ionic liquid-organic solvent as mobile phase. The effects of chromatographic columns, ultraviolet absorption reagents, imidazolium ILs, detection wavelength, organic solvents, pH values of the mobile phase and column temperature on the retention of morpholinium cations were investigated. The retention times of the cations were clearly decreased by the increase of the alkyl substituent length on imidazolium cation or with the increase of the concentration of 1-ethyl-3-methyl imidazolium tetrafluoroborate. The molecular structure of the anion of imidazolium IL which has UV absorption has influence on determination of the analytes. Under the optimal conditions, the detection limits (signal-to-noise ratio, S/N = 3) were 0.29 and 0.44 mg L-1 The method has been successfully applied to the determination of two ILs synthesized by chemistry laboratory. The method uses the liquid chromatography system, which is widely available in general laboratories, and the simple composition of mobile phase, thus make the quantitative analysis of no UV absorption morpholinium cations possible. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  16. Determination of Flow Rates in Capillary Liquid Chromatography Coupled to a Nanoelectrospray Source using Droplet Image Analysis Software.

    Science.gov (United States)

    Cohen, Alejandro M; Soto, Axel J; Fawcett, James P

    2016-08-02

    Liquid chromatography coupled to electrospray tandem mass spectrometry (LC-ESI-MS/MS) is widely used in proteomic and metabolomic workflows. Considerable analytical improvements have been observed when the components of LC systems are scaled down. Currently, nano-ESI is typically done at capillary LC flow rates ranging from 200 to 300 nL/min. At these flow rates, trouble shooting and leak detection of LC systems has become increasingly challenging. In this paper we present a novel proof-of-concept approach to measure flow rates at the tip of electrospray emitters when the ionization voltage is turned off. This was achieved by estimating the changes in the droplet volume over time using digital image analysis. The results are comparable with the traditional methods of measuring flow rates, with the potential advantages of being fully automatable and nondisruptive.

  17. Analysis of allergens in tubeimu saponin extracts by using rat basophilic leukemia 2H3 cell-based affinity chromatography coupled to liquid chromatography and mass spectrometry.

    Science.gov (United States)

    Zhang, Tao; Han, Shengli; Liu, Qi; Guo, Ying; He, Langchong

    2014-11-01

    An affinity two-dimensional chromatography method was developed for the recognition, separation, and identification of allergic components from tubeimu saponin extracts, a preparation often injected to treat various conditions as indicated by traditional Chinese medicine. Rat basophilic leukemia-2H3 cell membranes were used as the stationary phase of a membrane affinity chromatography column to capture components with affinity for mast cells that could be involved in a degranulation reaction. The retained components were enriched and analyzed by membrane affinity chromatography with liquid chromatography and mass spectrometry via a port switch valve. Suitability and reliability of the method was investigated using appropriate standards, and then, the method was applied to identify components retained from tubeimu saponin extracts. Tubeimoside A was identified in this way as a potential allergen, and degranulation assays confirmed that tubeimoside A induces RBL-2H3 cell degranulation in a dose-dependent manner. An increase in Ca(2+) influx indicated that degranulation induced by tubeimoside A is likely Ca(2+) dependent. Coupled with the degranulation assay, RBL-2H3 cell-based affinity chromatography coupled with liquid chromatography and mass spectrometry is an effective method for screening and identifying allergic components from tubeimu saponin extracts. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Two-dimensional preparative liquid chromatography system for preparative separation of minor amount components from complicated natural products

    Energy Technology Data Exchange (ETDEWEB)

    Qiu, Ying-Kun, E-mail: qyk@xmu.edu.cn; Chen, Fang-Fang; Zhang, Ling-Ling; Yan, Xia; Chen, Lin; Fang, Mei-Juan; Wu, Zhen, E-mail: wuzhen@xmu.edu.cn

    2014-04-01

    Highlights: • Preparative MDLC system was developed for separation of complicated natural products. • Medium-pressure LC and preparative HPLC were connected by interface of SPE. • Automated multi-step preparative separation of 25 compounds was achieved by using this system. - Abstract: An on-line comprehensive two-dimensional preparative liquid chromatography system was developed for preparative separation of minor amount components from complicated natural products. Medium-pressure liquid chromatograph (MPLC) was applied as the first dimension and preparative HPLC as the second one, in conjunction with trapping column and makeup pump. The performance of the trapping column was evaluated, in terms of column size, dilution ratio and diameter-height ratio, as well as system pressure from the view of medium pressure liquid chromatograph. Satisfactory trapping efficiency can be achieved using a commercially available 15 mm × 30 mm i.d. ODS pre-column. The instrument operation and the performance of this MPLC × preparative HPLC system were illustrated by gram-scale isolation of crude macro-porous resin enriched water extract of Rheum hotaoense. Automated multi-step preparative separation of 25 compounds, whose structures were identified by MS, {sup 1}H NMR and even by less-sensitive {sup 13}C NMR, could be achieved in a short period of time using this system, exhibiting great advantages in analytical efficiency and sample treatment capacity compared with conventional methods.

  19. Quantitative analysis of heterocyclic amines in urine by liquid chromatography coupled with tandem mass spectrometry.

    Science.gov (United States)

    Shin, Jeoung Hwa; Na, Yun-Cheol; Chung, Joo Hee; Gorinstein, Shela; Ahn, Yun Gyong

    2014-02-15

    A sensitive, reproducible, and rapid analytical method for the analysis of trace-level heterocyclic amines (HCAs) that are expected to have high levels of human exposure was developed. Liquid-liquid extraction (LLE) with dichloromethane (DCM) followed by solid-phase extraction (SPE) was carried out. Liquid extraction with DCM under basic conditions was efficient in extracting HCAs from urine samples. For further purification, mixed mode cationic exchange (MCX) cartridges were applied to eliminate the remaining interferences after liquid extraction. Separation and quantification were performed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) in selected reaction monitoring (SRM) mode. The overall recoveries ranged between 71.0% and 113.6% with relative standard deviations (RSDs) of 5.1% to 14.7% for the entire procedure. The limits of detection (LODs) and limits of quantification (LOQs) of the proposed analytical method were in the ranges of 0.04 to 0.10 ng/ml and 0.15 to 0.36 ng/ml, respectively. This method was applied to the analysis of monitoring in urine samples for Korean school children, and the results demonstrated that the method can be used for the trace determination of HCAs in urine samples. Copyright © 2013 Elsevier Inc. All rights reserved.

  20. Methods for Peptide and Protein Quantitation by Liquid Chromatography-Multiple Reaction Monitoring Mass Spectrometry*

    Science.gov (United States)

    Zhang, Haixia; Liu, Qinfeng; Zimmerman, Lisa J.; Ham, Amy-Joan L.; Slebos, Robbert J. C.; Rahman, Jamshedur; Kikuchi, Takefume; Massion, Pierre P.; Carbone, David P.; Billheimer, Dean; Liebler, Daniel C.

    2011-01-01

    Liquid chromatography-multiple reaction monitoring mass spectrometry of peptides using stable isotope dilution (SID) provides a powerful tool for targeted protein quantitation. However, the high cost of labeled peptide standards for SID poses an obstacle to multiple reaction monitoring studies. We compared SID to a labeled reference peptide (LRP) method, which uses a single labeled peptide as a reference standard for all measured peptides, and a label-free (LF) approach, in which quantitation is based on analysis of un-normalized peak areas for detected MRM transitions. We analyzed peptides from the Escherichia coli proteins alkaline phosphatase and β-galactosidase spiked into lysates from human colon adenocarcinoma RKO cells. We also analyzed liquid chromatography-multiple reaction monitoring mass spectrometry data from a recently published interlaboratory study by the National Cancer Institute Clinical Proteomic Technology Assessment for Cancer network (Addona et al. (2009) Nat. Biotechnol. 27: 633–641), in which unlabeled and isotopically labeled synthetic peptides or their corresponding proteins were spiked into human plasma. SID displayed the highest correlation coefficients and lowest coefficient of variation in regression analyses of both peptide and protein spike studies. In protein spike experiments, median coefficient of variation values were about 10% for SID and 20–30% for LRP and LF methods. Power calculations indicated that differences in measurement error between the methods have much less impact on measured protein expression differences than biological variation. All three methods detected significant (p proteins in a test set of 10 pairs of human lung tumor and control tissues. Further, the LRP and LF methods both detected significant differences (p quantitative liquid chromatography-multiple reaction monitoring mass spectrometry applications. PMID:21357624

  1. Analysis of recombinant Schistosoma mansoni antigen rSmp28 by on-line liquid chromatography-mass spectrometry combined with sodium dodecyl sulfate polyacrylamide gel electrophoresis

    NARCIS (Netherlands)

    Klarskov, K.; Roecklin, D.; Bouchon, B.; Sabatie, J.; Van Dorsselaer, A.; Bischoff, Rainer

    1994-01-01

    A recombinant Schistosoma mansoni antigen produced in Saccharomyces cerevisiae and purified by glutathione-Sepharose affinity chromatography was analyzed by tryptic peptide mapping using on-line reversed-phase high-performance liquid chromatography pneumatically assisted electrospray mass

  2. Determination of glycerol in oils and fats using liquid chromatography chloride attachment electrospray ionization mass spectrometry.

    Science.gov (United States)

    Jin, Chunfen; Viidanoja, Jyrki

    2017-01-15

    Existing liquid chromatography - mass spectrometry method for the analysis of short chain carboxylic acids was expanded and validated to cover also the measurement of glycerol from oils and fats. The method employs chloride anion attachment and two ions, [glycerol+35Cl]- and [glycerol+37Cl]-, as alternative quantifiers for improved selectivity of glycerol measurement. The averaged within run precision, between run precision and accuracy ranged between 0.3-7%, 0.4-6% and 94-99%, respectively, depending on the analyte ion and sample matrix. Selected renewable diesel feedstocks were analyzed with the method. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Chiral separation of pharmaceutical compounds using electrochemically modulated liquid chromatography (EMLC)

    Energy Technology Data Exchange (ETDEWEB)

    Wang, S. [Iowa State Univ., Ames, IA (United States)

    1999-02-12

    This research explores the application of a new technique, termed electrochemically modulated liquid chromatography (EMLC), to the chiral separations of pharmaceutical compounds. The introduction section provides a literature review of the technique and its applications, as well as brief overview of the research described in each of the next two chapters. Chapter 2 investigates the EMLC-based enantiomeric separation of a group of chiral benzodiazepines with β-cyclodextrin as a chiral mobile phase additive. Chapter 3 demonstrates the effects of several experimental parameters on the separation efficiency of drug enantiomers. The author concludes with a general summary and possible directions for future studies. Chapters 2 and 3 are processed separately.

  4. Detection of cheese whey and caseinomacropeptide in fermented milk beverages using high performance liquid chromatography

    OpenAIRE

    E.H.P. Andrade; M.R. Souza; L.M. Fonseca; C.F.A.M. Penna; M.M.O.P. Cerqueira; T. Roza; B. Seridan; M.F.S. Resende; F.A. Pinto; C.N.B.C. Villanoeva; M.O. Leite

    2014-01-01

    Cheese whey level and caseinomacropeptide (CMP) index of fermented milk beverages added with four levels of cheese whey (0, 10, 20, and 40%) and stored at 8-10oC for 0, 7, 14 and 21 days were determined by high performance liquid chromatography-gel filtration (HPLC-GF). Additionally, the interference of the starter culture and the storage time on the detection of cheese whey and CMP were investigated. Refrigerated storage up to 21 days did not affect (P>0.05) cheese whey and CMP amounts in mi...

  5. Increasing flexibility in two-dimensional liquid chromatography by pulsed elution of the first dimension

    DEFF Research Database (Denmark)

    Jakobsen, Simon S.; Christensen, Jan H.; Verdier, Sylvain

    2017-01-01

    This work demonstrates the development of an online two-dimensional liquid chromatography (2D-LC) method where the first dimension column is eluted by a sequence of pulses of increasing eluotropic strength generated by the LC pumps (pulsed-elution 2D-LC). Between the pulses, the first dimension...... online comprehensive 2D-LC: undersampling, difficulties in refocusing, and lack of flexibility in the selection of column dimensions and flow rates because the two dimensions constrain each other. The pulsed-elution 2D-LC was applied for the analysis of a basic fraction of vacuum gas oil. Peak capacity...

  6. Study of Saiga Horn Using High-Performance Liquid Chromatography with Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Kateřina Mikulíková

    2012-01-01

    Full Text Available The saiga horns have been investigated the using of modern analytic methods. High-performance liquid chromatography (HPLC with mass-spectrometric (MS and MS/MS detection and polyacrylamide gel electrophoresis (PAGE were used. It could be concluded that basic proteins of the saiga horns are keratins and collagen. The basic representation protein in all samples is keratin type I microfibrillar (from sheep, keratin type II microfibrillar (from sheep, collagen type I (α1 (from bovine and collagen type I (α2 (from bovine. Free amino acids we determined in all samples are nontreated by enzyme.

  7. Rapid confirmatory analysis of avermectin residues in milk by liquid chromatography tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Nurullah Ozdemir

    2016-01-01

    Full Text Available Our study developed a quick method for confirmatory analysis of avermectins (abamectin B1a, doramectin, ivermectin B1a, eprinomectin B1a, and moxidectin in bovine milk according to the European Commission Decision 2002/657/EC requirements. Avermectins were liquid–liquid extracted with acetonitrile, followed by an evaporation step, and then analyzed by liquid chromatography/electrospray ionization tandem mass spectrometry in the negative ion mode. An in-house method validation was performed and the data reported on specificity, linearity, recovery, limit of detection, limit of quantitation, decision limit, and detection capability. The advantage of this method is that low levels of avermectins are detectable and quantitatively confirmed at a rapid rate in milk.

  8. Novel materials and methods for solid-phase extraction and liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Ambrose, Diana [Iowa State Univ., Ames, IA (United States)

    1997-06-24

    This report contains a general introduction which discusses solid-phase extraction and solid-phase micro-extraction as sample preparation techniques for high-performance liquid chromatography, which is also evaluated in the study. This report also contains the Conclusions section. Four sections have been removed and processed separately: silicalite as a sorbent for solid-phase extraction; a new, high-capacity carboxylic acid functionalized resin for solid-phase extraction; semi-micro solid-phase extraction of organic compounds from aqueous and biological samples; and the high-performance liquid chromatographic determination of drugs and metabolites in human serum and urine using direct injection and a unique molecular sieve.

  9. Saffron authentication based on liquid chromatography high resolution tandem mass spectrometry and multivariate data analysis.

    Science.gov (United States)

    Rubert, Josep; Lacina, Ondrej; Zachariasova, Milena; Hajslova, Jana

    2016-08-01

    Saffron is one of the oldest and most expensive spices, which is often target of fraudulent activities. In this research, a new strategy of saffron authentication based on metabolic fingerprinting was developed. In the first phase, a solid liquid extraction procedure was optimized, the main aim was to isolate as maximal representation of small molecules contained in saffron as possible. In the second step, a detection method based on liquid chromatography coupled with high-resolution mass spectrometry was developed. Initially, principal component analysis (PCA) revealed clear differences between saffron cultivated and packaged in Spain, protected designation of origin (PDO), and saffron packaged in Spain of unknown origin, labeled Spanish saffron. Afterwards, orthogonal partial least square discriminant analysis (OPLS-DA) was favorably used to discriminate between Spanish saffron. The tentative identification of markers showed glycerophospholipids and their oxidized lipids were significant markers according to their origin. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Separative determination of ascorbic acid and erythorbic acid by high-performance liquid chromatography.

    Science.gov (United States)

    Arakawa, N; Otsuka, M; Kurata, T; Inagaki, C

    1981-01-01

    High-performance liquid chromatography (HPLC) was applied to the determination of ascorbic acid (AsA) and erythorbic acid (ErA). The apparatus was a Shimadzu model LC-2P Liquid Chromatograph equipped with a UV detector set at 254 nm. The separation was achieved on a LiChrosorb-NH2 column which was pre-treated with 0.1 M ammonium monophosphate solution using a mixture of acetonitrile, acetic acid and water (87:2:11, v/v) as an eluant. The HPLC method has the following advantages: AsA and ErA are quantitated after being distinctly separated, analysis time per one sample is short, and AsA or ErA levels as low as 1.0 X 10(-2) microgram are detectable. Recovery experiments with dehydro-AsA and dehydro-ErA, involving reduction with H2S, give satisfactory results.

  11. Quantitative analysis of tetramethylenedisulfotetramine (tetramine) spiked into beverages by liquid chromatography-tandem mass spectrometry with validation by gas chromatography-mass spectrometry.

    Science.gov (United States)

    Owens, Janel; Hok, Saphon; Alcaraz, Armando; Koester, Carolyn

    2009-05-27

    Tetramethylenedisulfotetramine, commonly known as tetramine, is a highly neurotoxic rodenticide (human oral LD(50) = 0.1 mg/kg) used in hundreds of deliberate and accidental food poisoning events in China. This paper describes a method for the quantitation of tetramine spiked into beverages, including milk, juice, tea, cola, and water, with cleanup by C8 solid phase extraction and liquid-liquid extraction. Quantitation by high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) was based upon fragmentation of m/z 347 to m/z 268. The method was validated by gas chromatography-mass spectrometry (GC-MS) operated in selected ion monitoring mode for ions m/z 212, 240, and 360. The limit of quantitation was 0.10 μg/mL by LC-MS/MS versus 0.15 μg/mL for GC-MS. Fortifications of the beverages at 2.5 and 0.25 μg/mL were recovered ranging from 73 to 128% by liquid-liquid extraction for GC-MS analysis, from 13 to 96% by SPE, and from 10 to 101% by liquid-liquid extraction for LC-MS/MS analysis.

  12. Quantitative Analysis of Tetramethylenedisulfotetramine ("Tetramine") Spiked into Beverages by Liquid Chromatography Tandem Mass Spectrometry with Validation by Gas Chromatography Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Owens, J; Hok, S; Alcaraz, A; Koester, C

    2008-11-13

    Tetramethylenedisulfotetramine, commonly known as tetramine, is a highly neurotoxic rodenticide (human oral LD{sub 50} = 0.1 mg/kg) used in hundreds of deliberate food poisoning events in China. Here we describe a method for quantitation of tetramine spiked into beverages, including milk, juice, tea, cola, and water and cleaned up by C8 solid phase extraction and liquid-liquid extraction. Quantitation by high performance liquid chromatography tandem mass spectrometry (LC/MS/MS) was based upon fragmentation of m/z 347 to m/z 268. The method was validated by gas chromatography mass spectrometry (GC/MS) operated in SIM mode for ions m/z 212, 240, and 360. The limit of quantitation was 0.10 {micro}g/mL by LC/MS/MS versus 0.15 {micro}g/mL for GC/MS. Fortifications of the beverages at 2.5 {micro}g/mL and 0.25 {micro}g/mL were recovered ranging from 73-128% by liquid-liquid extraction for GC/MS analysis, 13-96% by SPE and 10-101% by liquid-liquid extraction for LC/MS/MS analysis.

  13. A comparison of frontal and nonfrontal methods for determining solid-liquid adsorption isotherms using inverse liquid chromatography.

    Science.gov (United States)

    Ylä-Mäihäniemi, Pirre P; Williams, Daryl R

    2007-03-27

    Inverse liquid chromatography (ILC) has been used to determine experimental isotherms for the equilibrium adsorption of cyclohexanone onto a silica (61.8 m(2)/g) from hexane using the peak maximum (PM), elution by characteristic point (ECP), frontal analysis (FA), and frontal analysis by characteristic point (FACP) methods. Isotherms obtained using these four approaches gave good internal agreement, as well as being in good agreement with classically determined isotherms. Columns were successfully packed using a dry powder packing method with 9 microm diameter silica particles, and excellent intercolumn and instrument to instrument reproducibility was obtained for PM isotherms. The theoretical background to the PM, ECP, FA, and FACP methods, as well as the practical facets of isotherm determination using these methods, is outlined in this work.

  14. Comparison of microemulsion electrokinetic chromatography with high-performance liquid chromatography for fingerprint analysis of resina draconis.

    Science.gov (United States)

    Cao, Yuhua; Gong, Wenjun; Li, Nan; Yin, Changna; Wang, Yun

    2008-11-01

    Microemulsion electrokinetic chromatography (MEEKC) has been developed for fingerprint analysis of resina draconis, a substitute for sanguis draconis in the Chinese market. The microemulsion as the running buffer was made up of 3.3% (w/v) sodium dodecyl sulfate (SDS), 6.6% (w/v) n-butanol, 0.8% (w/v) n-octane, and 10 mmol/L sodium tetraborate buffer (pH 9.2), which was also used as the solvent for ultrasonic extraction of both water- and fat-soluble compounds in the traditional Chinese medicine samples. Four batches of resina draconis obtained from different pharmaceutical factories located in different geographic regions were used to establish the electrophoretic fingerprint. MEEKC was performed using a Beckman PACE/MDQ system equipped with a diode-array detector and with monitoring at 280 nm. The fingerprint of resina draconis comprised 27 common peaks within 100 min. The relative standard deviations of the relative migration time of these common peaks were less than 2.1%. Through repetitive injection of the sample solution six times in 24 h, all relative standard deviations of the migration time and peak area of loureirin A and loureirin B were less than 2.5 and 3.8%, which demonstrated that the method had good stability and reproducibility. The relative peak areas of these common peaks in the electropherograms of four batches of resina draconis were processed with two mathematical methods, the correlation coefficient and the interangle cosine, to valuate the similarity. The values of the similarity degree of all samples were more than 0.91, which showed resina draconis samples from different origins were consistent. On the other hand, high-performance liquid chromatography (HPLC) coupled with photodiode-array detection was also applied to establish the fingerprint of resina draconis. The samples were separated with a LiChrospher C(18) column using acetonitrile (solvent A) and water containing 0.1% H(3)PO(4) (solvent B) as the mobile phase in linear gradient

  15. Recent trends and applications in liquid-phase chromatography enantioseparation of atropisomers.

    Science.gov (United States)

    Peluso, Paola; Mamane, Victor; Aubert, Emmanuel; Cossu, Sergio

    2017-08-01

    The term Atropisomerism defines a type of enantiomerism that arises from hindered rotation around an axis that makes two rotational isomers enantiomers. Atropisomeric compounds are present in nature, being important building blocks of many biologically active compounds. Atropisomers have been extensively studied in environmental and toxicological chemistry and, in particular, separating them has become a need in specific fields as organic synthesis and pharmaceutical research. High-performance LC has been fruitfully applied in this context, and despite HPLC separation on chiral stationary phase is considered as mature technology nowadays, in the last years several advancements and applications contributed to study compounds where axial chirality is of utmost importance. Moreover, dynamic chromatography has been exploited as a versatile tool for kinetic studies of systems with slow internal motion gaining essential information on their stereodynamic behavior. On this basis, current topics and trends in liquid-phase chromatography enantioseparation of atropisomers are presented herein with specific focus on new representative applications with HPLC, covering years 2010-2016. Some examples concerning supercritical fluid chromatography applications are also reported. This review aims to provide the reader with a modern overview of the field to highlight the renewed interest toward the chemistry of molecules with atropisomeric properties. A systematic compilation of all published literature has not been attempted. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Screening of drugs and toxic compounds with liquid chromatography-linear ion trap tandem mass spectrometry.

    Science.gov (United States)

    Sauvage, François-Ludovic; Saint-Marcoux, Franck; Duretz, Bénédicte; Deporte, Didier; Lachatre, Gérard; Marquet, Pierre

    2006-09-01

    In clinical and forensic toxicology, general unknown screening is used to detect and identify exogenous compounds. In this study, we aimed to develop a comprehensive general unknown screening method based on liquid chromatography coupled with a hybrid triple-quadrupole linear ion trap mass spectrometer. After solid-phase extraction, separation was performed using gradient reversed-phase chromatography. The mass spectrometer was operated in the information-dependent acquisition mode, switching between a survey scan acquired in the Enhanced Mass Spectrometry mode with dynamic subtraction of background noise and a dependent scan obtained in the enhanced product ion scan mode. The complete cycle time was 1.36 s. A library of 1000 enhanced product ion-tandem mass spectrometry spectra in positive mode and 250 in negative mode, generated using 3 alternated collision tensions during each scan, was created by injecting pure solutions of drugs and toxic compounds. Comparison with HPLC-diode array detection and gas chromatography-mass spectrometry for the analysis of 36 clinical samples showed that linear ion trap tandem mass spectrometry could identify most of the compounds (94% of the total). Some compounds were detected only by 1 of the other 2 techniques. Specific clinical cases highlighted the advantages and limitations of the method. A unique combination of new operating modes provided by hybrid triple-quadrupole linear ion trap mass spectrometers and new software features allowed development of a comprehensive and efficient method for the general unknown screening of drugs and toxic compounds in blood or urine.

  17. Characterization of peak capacity of microbore liquid chromatography columns using gradient kinetic plots.

    Science.gov (United States)

    Hetzel, Terence; Blaesing, Christina; Jaeger, Martin; Teutenberg, Thorsten; Schmidt, Torsten C

    2017-02-17

    The performance of micro-liquid chromatography columns with an inner diameter of 0.3mm was investigated on a dedicated micro-LC system for gradient elution. Core-shell as well as fully porous particle packed columns were compared on the basis of peak capacity and gradient kinetic plot limits. The results for peak capacity showed the superior performance of columns packed with sub-2μm fully porous particles compared to 3.0μm fully porous and 2.7μm core-shell particles within a range of different gradient time to column void time ratios. For ultra-fast chromatography a maximum peak capacity of 16 can be obtained using a 30s gradient for the sub-2μm fully porous particle packed column. A maximum peak capacity of 121 can be achieved using a 5min gradient. In addition, the influence of an alternative detector cell on the basis of optical waveguide technology and contributing less to system variance was investigated showing an increased peak capacity for all applied gradient time/column void time ratios. Finally, the influence of pressure was evaluated indicating increased peak capacity for maximum performance whereas a limited benefit for ultra-fast chromatography with gradient times below 30s was observed. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Microfluidic platform for liquid chromatography-tandem mass spectrometry analyses of complex peptide mixtures.

    Science.gov (United States)

    Xie, Jun; Miao, Yunan; Shih, Jason; Tai, Yu-Chong; Lee, Terry D

    2005-11-01

    A microfluidic chip that integrates all the fluidic components of a gradient liquid chromatography (LC) system is described. These chips were batch-fabricated on a silicon wafer using photolithographic processes and with Parylene as the main structural material. The fabricated chip includes three electrolysis-based electrochemical pumps, one for loading the sample and the other two for delivering the solvent gradient; platinum electrodes for delivering current to the pumps and establishing the electrospray potential; a low-volume static mixer; a column packed with silica-based reversed-phase support; integrated frits for bead capture; and an electrospray nozzle. The fabricated structures were able to withstand pressures in excess of 250 psi. The device was used to perform a liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of a mixture of peptides from the trypsin digestion of bovine serum albumen (BSA). Gradient elution through the 1.2-cm column was performed at a flow rate of 80 nL/min. Compared to the analysis of the same sample using a commercial nanoflow LC system, the chromatographic resolution was nearly as good, and the total cycle time was significantly reduced because of the minimal volume between the pumps and the column. Results demonstrate the potential of mass-produced, low-cost microfluidic systems capable of performing LC separations for proteomics applications.

  19. Comparison of capillary electrophoresis and high performance liquid chromatography methods for caffeine determination in decaffeinated coffee

    Directory of Open Access Journals (Sweden)

    Carolina Schaper Bizzotto

    2013-03-01

    Full Text Available Decaffeinated coffee accounts for 10 percent of coffee sales in the world; it is preferred by consumers that do not wish or are sensitive to caffeine effects. This article presents an analytical comparison of capillary electrophoresis (CE and high performance liquid chromatography (HPLC methods for residual caffeine quantification in decaffeinated coffee in terms of validation parameters, costs, analysis time, composition and treatment of the residues generated, and caffeine quantification in 20 commercial samples. Both methods showed suitable validation parameters. Caffeine content did not differ statistically in the two different methods of analysis. The main advantage of the high performance liquid chromatography (HPLC method was the 42-fold lower detection limit. Nevertheless, the capillary electrophoresis (CE detection limit was 115-fold lower than the allowable limit by the Brazilian law. The capillary electrophoresis (CE analyses were 30% faster, the reagent costs were 76.5-fold, and the volume of the residues generated was 33-fold lower. Therefore, the capillary electrophoresis (CE method proved to be a valuable analytical tool for this type of analysis.

  20. Core-shell microspheres with porous nanostructured shells for liquid chromatography.

    Science.gov (United States)

    Ahmed, Adham; Skinley, Kevin; Herodotou, Stephanie; Zhang, Haifei

    2018-01-01

    The development of new stationary phases has been the key aspect for fast and efficient high-performance liquid chromatography separation with relatively low backpressure. Core-shell particles, with a solid core and porous shell, have been extensively investigated and commercially manufactured in the last decade. The excellent performance of core-shell particles columns has been recorded for a wide range of analytes, covering small and large molecules, neutral and ionic (acidic and basic), biomolecules and metabolites. In this review, we first introduce the advance and advantages of core-shell particles (or more widely known as superficially porous particles) against non-porous particles and fully porous particles. This is followed by the detailed description of various methods used to fabricate core-shell particles. We then discuss the applications of common silica core-shell particles (mostly commercially manufactured), spheres-on-sphere particles and core-shell particles with a non-silica shell. This review concludes with a summary and perspective on the development of stationary phase materials for high-performance liquid chromatography applications. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Hb A1c Separation by High Performance Liquid Chromatography in Hemoglobinopathies

    Directory of Open Access Journals (Sweden)

    Vani Chandrashekar

    2016-01-01

    Full Text Available Hb A1c measurement is subject to interference by hemoglobin traits and this is dependent on the method used for determination. In this paper we studied the difference between Hb A1c measured by HPLC in hemoglobin traits and normal chromatograms. We also studied the correlation of Hb A1c with age. Hemoglobin analysis was carried out by high performance liquid chromatography. Spearman’s rank correlation was used to study correlation between A1c levels and age. Mann-Whitney U test was used to study the difference in Hb A1c between patients with normal hemoglobin and hemoglobin traits. A total of 431 patients were studied. There was positive correlation with age in patients with normal chromatograms only. No correlation was seen in Hb E trait or beta thalassemia trait. No significant difference in Hb A1c of patients with normal chromatograms and patients with hemoglobin traits was seen. There is no interference by abnormal hemoglobin in the detection of A1c by high performance liquid chromatography. This method cannot be used for detection of A1c in compound heterozygous and homozygous disorders.

  2. Analysis of glyphosate residues in cereals using liquid chromatography-mass spectrometry (LC-MS/MS)

    DEFF Research Database (Denmark)

    Granby, Kit; Johannesen, S.; Gabrielsen, Martin Vahl

    2003-01-01

    A fast and specific method for the determination of glyphosate in cereals is described. The method is based on extraction with water by ultrasonication. The samples are cleaned up and separated by high-performance liquid chromatography on a polystyrene-based reverse-phase column (clean-up) in ser......A fast and specific method for the determination of glyphosate in cereals is described. The method is based on extraction with water by ultrasonication. The samples are cleaned up and separated by high-performance liquid chromatography on a polystyrene-based reverse-phase column (clean...... monitored m/z 168--> 150 (glyphosate) and 170-->152 (internal standard 2- 13 (CN)-N-15-glyphosate) for quantification. The mean recovery was 85% ( n =32) at spiking levels from 0.03 to 0.33 mg kg(-1) . From 1998 to 2001, from the analysis of about 50 samples per annum, a reduction in the glyphosate residues...... was observed owing to a Danish trade decision not to use grain with glyphosate residues for milling or bread production....

  3. High-performance liquid chromatography mass spectrometry of gold and alloy clusters protected by hydrophilic thiolates.

    Science.gov (United States)

    Niihori, Yoshiki; Shima, Daisuke; Yoshida, Kana; Hamada, Kota; Nair, Lakshmi V; Hossain, Sakiat; Kurashige, Wataru; Negishi, Yuichi

    2018-01-25

    In this work, we found two hydrophilic interaction liquid chromatography (HILIC) columns for high-performance liquid chromatography (HPLC) suitable for the high-resolution separation of hydrophilic metal clusters. The mass distributions of the product mixtures of hydrophilic metal clusters were evaluated via HPLC mass spectrometry (LC/MS) using these HILIC columns. Consequently, we observed multiple clusters that had not been previously reported for glutathionate (SG)-protected gold clusters (Aun(SG)m). Additionally, we demonstrated that Aun-xMx(SG)m alloy clusters (M = Ag, Cu, or Pd) in which part of the Au in the Aun(SG)m cluster is replaced by a heteroelement can be synthesized, similar to the case of hydrophobic alloy clusters. It is easy to evaluate the mass distributions of hydrophilic metal clusters using this method. Thus, remarkable progress in the synthesis techniques of hydrophilic metal clusters through the use of this method is anticipated, as is the situation for hydrophobic metal clusters.

  4. Determination of five bisphenols in commercial milk samples by liquid chromatography coupled to fluorescence detection.

    Science.gov (United States)

    Grumetto, Lucia; Gennari, Oriella; Montesano, Domenico; Ferracane, Rosalia; Ritieni, Alberto; Albrizio, Stefania; Barbato, Francesco

    2013-09-01

    The presence of five bisphenols, i.e., bisphenol F, bisphenol A, bisphenol B, bisphenol F diglycidyl ether, and bisphenol A diglycidyl ether, was monitored in commercial milk packed in plastic bottles marketed in Italy. The new validated method includes a solid-phase extraction procedure followed by liquid chromatography with fluorescence detection. All positive results were confirmed by liquid chromatography-tandem mass spectrometry analysis. The limits of detection and quantification and the recovery percentages indicated that the method is suitable for detecting bisphenols in milk at concentrations far below the legal limits. Of 68 commercial milk samples analyzed, no bisphenol was found in 27 samples (39.7%), and 41 samples (60.3%) contained one or more bisphenols. The bisphenol most frequently found was bisphenol F (36 samples, 52.9%) followed by bisphenol A (20 samples, 29.4%) and bisphenol B (6 samples, 8.8%). Taking into consideration the limits of detection, no sample contained either bisphenol F diglycidyl ether or bisphenol A diglycidyl ether.

  5. Capillary Liquid Chromatography Mass Spectrometry Analysis of Intact Monolayer-Protected Gold Clusters in Complex Mixtures.

    Science.gov (United States)

    Black, David M; Bach, Stephan B H; Whetten, Robert L

    2016-06-07

    In some respects, large noble-metal clusters protected by thiolate ligands behave as giant molecules of definite composition and structure; however, their rigorous analysis continues to be quite challenging. Analysis of complex mixtures of intact monolayer-protected clusters (MPCs) by liquid chromatography mass spectrometry (LC-MS) could provide quantitative identification of the various components present. This advance is critical for biomedical and toxicological research, as well as in fundamental studies that rely on the identification of selected compositions. This work expands upon the separate LC and MS results previously achieved, by interfacing the capillary liquid chromatograph directly to the electrospray source of the mass spectrometer, in order to provide an extremely sensitive, quantitative, and rapid means to characterize MPCs and their derivatives far beyond that of earlier reports. Here, we show that nonaqueous reversed-phase chromatography can be coupled to mass-spectrometry detection to resolve complex mixtures in minute (∼100 ng) samples of gold MPCs, of molecular masses up to ∼40 kDa, and with single-species sensitivity easily demonstrated for components on the level of sub-10 ng or picomole (1 pmol).

  6. Metabolic profiling assisted quality assessment of Rhodiola rosea extracts by high-performance liquid chromatography.

    Science.gov (United States)

    Wang, Zhanguo; Hu, Huiling; Chen, Fang; Zou, Liang; Yang, Mingfu; Wang, Anqi; Foulsham, James E; Lan, Ke

    2012-05-01

    In this work, fast and sensitive high-performance liquid chromatography (HPLC) coupled with multivariate analysis was utilized to assist the quality assessment of Rhodiola rosea extracts (RREs). 131 peaks were separated and detected in RREs on a fused-core C18 column. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) of the chromatographic data demonstrated that 10 batches of RREs could be well-differentiated and categorized into three groups which were closely related to the origins of RREs. Partial least square-discriminant analysis (PLS-DA) showed that the quality differentiation might be explained by at least 6 components, in which rosavin was characterized by an external reference, rosiridine was identified by liquid chromatography-mass spectrometry (LC-MS), and the mass spectra of the others were provided. The observation that the level of rosavin was more relevant to the multivariate chromatographic data than the ones of salidroside and tyrosol, the other two components commonly used to standardize RREs, was confirmed by the PLS prediction models. Results of the present study not only indicated that rosavin was a rational marker to represent the quality of RREs, but also demonstrated the power of HPLC-based metabolic profiling in the quality assessment of herbal extracts. © Georg Thieme Verlag KG Stuttgart · New York.

  7. [Estimation of chloramphenicol in the working area air by high performance liquid chromatography].

    Science.gov (United States)

    Kristova-Bagdasarian, V L; Chokhadzhieva, D

    2008-01-01

    Chloramphenicol (levomycetin) is a broad-spectrum antibiotic that is active against gram-positive and gram-negative microorganisms. At present, it is manufactured via organic synthesis. Working place air becomes polluted during the manufacture and use of medicines containing chloramphenicol. In the working place air, chloramphenicol is present as a disintegration aerosol and may provoke occupational diseases of varying severity in the exposed persons. A procedure has been determined to measure air chloramphenicol, by using high performance liquid chromatography. Aspiration through an AFA FPP-15 aerosol filter is a suitable device for air chloramphenicol sampling. The selected chloramphenicol is removed from the filter via triple methanol extraction in an ultrasound bath. The pooled extract is evaporated to dryness in a current of nitrogen and the dry residue is dissolved in the mobile phase containing acetonitrile : buffer (pH 4.8) = 30:70. The chloramphenicol determination procedure using reverse-phase liquid chromatography with ultraviolet detection at a wavelength of 275 nm has been developed and completely validated. Chromatographic conditions are given. The retention time of chloramphenicol is 6.5 min. The detection limit is 0.1 microg/cm3. The method is noted for a linear relationship between the concentration of chloramphenicol (microg/cm3) and the peak area (mm2) in the range of 1 to 20 microg/cm3.

  8. Validation of a technique by high-performance liquid chromatography for the determination of total isoflavones

    Directory of Open Access Journals (Sweden)

    Pilar A. Soledispa Cañarte

    2017-04-01

    Full Text Available Context: Isoflavones may act as selective regulators in the prevention of various diseases. The most important source of isoflavones is the soy, from which different phytotherapeutics are elaborated of use in Ecuadorian population. However, its concentration varies depending on several factors, therefore quality assessment need to be carried out through out several analytical methods. Aims: To validate an analytical method by high precision liquid chromatography (HPLC to quantify total isoflavones in herbal medicine. Methods: To quantify isoflavones, it was used a brand liquid chromatography with UV/VIS detector at 260 nm, C-18 column using isocratic method. The mobile phase was composed of 2% acetic acid: acetonitrile (75:25. The quantification was performed against reference standard. The parameters for the validation followed the established in the USP 33. Results: The chromatogram presented six peaks with elution between 1.557 and 18.913 min. The linearity of the system and the method got r2 equal to 0.98 and 0.99 respectively. The coefficients of variation 1.5% in the study of repetitiveness and 2% in intermediate precision. The accuracy of the adjusted lineal model exhibited r=0.95 and intercept reliable interval (-0.921; 1.743. Conclusions: The validated method was specific, accurate, precise and linear. It can be used for quality control and stability studies of isoflavones present in herbal medicine.

  9. Determination of bromate in drinking water by ultraperformance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Alsohaimi, Ibrahim Hotan; Alothman, Zeid Abdullah; Khan, Mohammad Rizwan; Abdalla, Mohammad Abulhassan; Busquets, Rosa; Alomary, Ahmad Khodran

    2012-10-01

    Bromate is a byproduct formed as a result of disinfection of bromide-containing source water with ozone or hypochlorite. The International Agency for Research on Cancer has recognized bromate as a possible human carcinogen, thus it is essential to determine in drinking water. Present work highlights a development of sensitive and fast analytical method for bromate determination in drinking water by using ultraperformance liquid chromatography-tandem mass spectrometry. The quality parameters of the developed method were established, obtaining very low limit of detection (0.01 ng/mL), repeatability and reproducibility have been found to be less than 3% in terms of relative standard deviation when analyzing a bromate standard at 0.05 μg/mL with 0.4 min analysis time. Developed method was applied for the analysis of metropolitan and bottled water from Saudi Arabia; 22 samples have been analyzed. Bromate was detected in the metropolitan water samples (from desalinization source) at concentrations ranging between 3.43 and 75.04 ng/mL and in the bottled water samples at concentrations ranging between 2.07 and 21.90 ng/mL. Moreover, in comparison to established analytical methods such as liquid chromatography-tandem mass spectrometry, the proposed method was found to be very sensitive, selective and rapid for the routine analysis of bromate at low level in drinking water. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. DETECTION AND QUANTIFICATION OF PHYTOCHEMICAL MARKERS OF Ilex paraguariensis BY LIQUID CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    Rodrigo M. C. Pinto

    2015-11-01

    Full Text Available Ilex paraguariensis (yerba-mate is used as a beverage, and its extract requires adequate quality control methods in order to guarantee quality and safe use. Strategies to develop and optimize a chromatographic method to quantify theobromine, caffeine, and chlorogenic acid in I. paraguariensis extracts were evaluated by applying a quality by design (QbD model and ultra high-performance liquid chromatography (UHPLC. The presence of these three phytochemical markers in the extracts was evaluated using UHPLC-MS and was confirmed by the chromatographic bands in the total ion current traces (m/z of 181.1 [M+H]+, 195.0 [M+H]+, and 353.0 [M−H]−, respectively. The developed method was then transferred to a high-performance liquid chromatography (HPLC platform, and the three phytochemical markers were used as external standards in the validation of a method for analyses of these compounds in extracts using a diode array detector (DAD. The validated method was applied to quantify the chlorogenic acid, caffeine, and theobromine in the samples. HPLC-DAD chromatographic fingerprinting was also used in a multivariate approach to process the entire data and to separate the I. paraguariensis extracts into two groups. The developed method is very useful for qualifying and quantifying I. paraguariensis extracts.

  11. [Determination of pyriproxyfen residue in vegetables and fruits by liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Zhao, Guangyu; Guo, Dehua; Zhao, Shanzhen; Sheng, Yonggang; Deng, Xiaojun

    2009-07-01

    A method of liquid chromatography-tandem mass spectrometry (LC-MS/MS) was established for the determination of pyriproxyfen residue in vegetables and fruits. The pyriproxyfen residue in the sample was extracted by acidified acetonitrile with the presence of sodium acetate buffer, and cleaned up by PSA (primary secondary amine) sorbent, and the separation was performed by ultrafast liquid chromatography (UFLC) on a CAPCELL PAK C18 column (50 mm x 2.0 mm, 3 microm) and the gradient elution of acetonitrile (containing 0.1% formic acid) and 2 mmol/L ammonium acetate solution (containing 0.1% formic acid) at a flow rate of 0.3 mL/min. The calibration curve was linear between the peak area and the con centration of 2.5 - 50 microg/L with the correlation coefficient more than 0.9999. The limit of quantification of pyriproxyfen was 5 microg/kg. The average recoveries spiked at three concentrations of 5, 50 and 100 microg/kg ranged 84.7% -91.5%. The relative standard deviations (n = 10) were all less than 10%. The method is selective without interference and suitable for the determination of pyriproxyfen residue in vegetables and fruits.

  12. Separation of tryptophan enantiomers by ligand-exchange chromatography with novel chiral ionic liquids ligand.

    Science.gov (United States)

    Qing, Haiqun; Jiang, Xinyu; Yu, Jingang

    2014-03-01

    Chiral ionic liquids (CILs) with amino acids as cations have been applied as novel chiral ligands coordinated with Cu(2+) to separate tryptophan enantiomers in ligand exchange chromatography. Four kinds of amino acid ionic liquids, including [L-Pro][CF3COO], [L-Pro][NO3], [L-Pro]2[SO4], and [L-Phe][CF3COO] were successfully synthesized and used for separation of tryptophan enantiomers. To optimize the separation conditions, [L-Pro][CF3COO] was selected as the model ligand. Some factors influencing the efficiency of chiral separation, such as copper ion concentration, CILs concentration, methanol ratio (methanol/H2O, v/v), and pH, were investigated. The obtained optimal separation conditions were as follows: 8.0 mmol/L Cu(OAc)2, 4.0 mmol/L [L-Pro][CF3COO], and 20% (v/v) methanol at pH 3.6. Under the optimum conditions, acceptable enantioseparation of tryptophan enantiomers could be observed with a resolution of 1.89. The results demonstrate the good applicability of CILs with amino acids as cations for chiral separation. Furthermore, a comparative study was also conducted for exploring the mechanism of the CILs as new ligands in ligand exchange chromatography. © 2014 Wiley Periodicals, Inc.

  13. Development and application of liquid chromatography coupled to isotope ratio mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Lijun

    2014-02-19

    Stable isotope analysis has found widespread applications in various disciplines such as archaeology, geochemistry, biology, food authenticity, and forensic science. Coupling chromatography to isotope ratio mass spectrometry for compound-specific isotope analysis (CSIA) is a trend, as it provides several advantages over bulk isotope analysis, e.g., relatively simple sample preparation, the ability to measure individual compounds in a complex mixture in one run, and the reduced sample size required for precise isotope analysis. Gas chromatography coupled to isotope ratio mass spectrometry (GC/IRMS) has been well-established for compound-specific isotope analysis of volatile organic compounds within the last two decades. However, an interface combining liquid chromatography with isotope ratio mass spectrometry (LC/IRMS) was not commercially available until 2004. The current design of the interface requires using a carbon-free eluent in chromatographic separation. This requirement limits the application of the most frequently used reversed-phase liquid chromatography in CSIA, because the elution strength of water at room temperature is too low to serve as mobile phase in reversed-phase separations. In order to increase the elution strength of water, we propose using high temperature water for chromatographic elution. The polarity of water decreases with an increase of temperature, yielding increased elution strength in reversed-phase columns. Therefore, high temperature water can be used as eluent instead of organic solvent for combining reversed-phase liquid chromatography with isotope ratio mass spectrometry (RPLC/IRMS). Additionally, temperature gradients can replace organic solvent gradients to increase chromatographic resolution. This is very important for LC/IRMS analysis, as precise isotope analysis requires baseline separation of analytes. In this thesis, high-temperature reversed-phase liquid chromatography was coupled to, and for the first time carefully

  14. Quantification of Quercetin and Rutin from Benincasa hispida Seeds and Carissa Congesta Roots by High-performance Thin Layer Chromatography and High-performance Liquid Chromatography.

    Science.gov (United States)

    Doshi, Gaurav Mahesh; Une, Hemant Devidas

    2016-01-01

    In Indian Ayurvedic system, Benincasa hispida (BH) and Carissa congesta (CC) are well-known plants used for major and minor ailments. BH has been regarded as Kushmanda, whereas CC has been used in immune-related disorders of the human system. Quercetin and rutin identified from the vast plethora of plant extracts have proved to possess ethnopharmacological relevance. In present studies, we have determined quercetin and rutin in terms of percentage in BH seeds and CC roots by high-performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC). After extraction and phytochemical screening, the extracts were subjected to quantification for the presence of quercetin and rutin by HPTLC and HPLC. HPTLC showed quercetin as 44.60, 27.13% and rutin as 32.00, 36.31% w/w, whereas HPLC revealed quercetin as 34.00, 35.00% and rutin as 21.99, 45.03% w/v in BH and CC extracts, respectively. The BH and CC extracts have elucidated peaks that were corresponding with standard peaks on undertaking chromatographic studies. Quercetin and rutin are isolated from BH seeds and CC roots by High Performance. Thin Layer Chromatography and High Performance Liquid Chromatography. HPTLC revealed presence of quercetin as 44.60, 27.13 % and rutin as 32.00, 36.31 % w/w. HPLC revealed presence of quercetin as 34.00, 35.00 % and rutin as 21.99, 45.03 % w/v. Abbreviation Used: HPTLC: High Performance Thin Layer Chromatography; HPLC: High Pressure Liquid Chromatography, UV: Ultraviolet, CC: Carissa congesta, BH: Benincasa hispida.

  15. LC-MSsim – a simulation software for liquid chromatography mass spectrometry data

    Directory of Open Access Journals (Sweden)

    Gröpl Clemens

    2008-10-01

    Full Text Available Abstract Background Mass Spectrometry coupled to Liquid Chromatography (LC-MS is commonly used to analyze the protein content of biological samples in large scale studies. The data resulting from an LC-MS experiment is huge, highly complex and noisy. Accordingly, it has sparked new developments in Bioinformatics, especially in the fields of algorithm development, statistics and software engineering. In a quantitative label-free mass spectrometry experiment, crucial steps are the detection of peptide features in the mass spectra and the alignment of samples by correcting for shifts in retention time. At the moment, it is difficult to compare the plethora of algorithms for these tasks. So far, curated benchmark data exists only for peptide identification algorithms but no data that represents a ground truth for the evaluation of feature detection, alignment and filtering algorithms. Results We present LC-MSsim, a simulation software for LC-ESI-MS experiments. It simulates ESI spectra on the MS level. It reads a list of proteins from a FASTA file and digests the protein mixture using a user-defined enzyme. The software creates an LC-MS data set using a predictor for the retention time of the peptides and a model for peak shapes and elution profiles of the mass spectral peaks. Our software also offers the possibility to add contaminants, to change the background noise level and includes a model for the detectability of peptides in mass spectra. After the simulation, LC-MSsim writes the simulated data to mzData, a public XML format. The software also stores the positions (monoisotopic m/z and retention time and ion counts of the simulated ions in separate files. Conclusion LC-MSsim generates simulated LC-MS data sets and incorporates models for peak shapes and contaminations. Algorithm developers can match the results of feature detection and alignment algorithms against the simulated ion lists and meaningful error rates can be computed. We

  16. Simultaneous determination of niacin and pyridoxine at trace levels by using diode array high-performance liquid chromatography and liquid chromatography with quadrupole time-of-flight tandem mass spectrometry.

    Science.gov (United States)

    Sel, Sabriye; Öztürk Er, Elif; Bakırdere, Sezgin

    2017-10-26

    A highly sensitive and simple diode-array high-performance liquid chromatography and liquid chromatography with quadrupole time-of-flight tandem mass spectrometry method was developed for the simultaneous determination of niacin and pyridoxine in pharmaceutical drugs, tap water and wastewater samples. To determine the in vivo behavior of niacin and pyridoxine, analytes were subjected to simulated gastric conditions. The calibration plots of the diode-array high-performance liquid chromatography and liquid chromatography with quadrupole time-of-flight tandem mass spectrometry method showed good linearity over a wide concentration range with close to 1.0 correlation coefficients for both analytes. The limit of detection/limit of quantitation values for liquid chromatography quadrupole time-of-flight tandem mass spectrometry analysis were 1.98/6.59 and 1.3/4.4 μg L-1 for niacin and pyridoxine, respectively, while limit of detection/limit of quantitation values for niacin and pyridoxine in high-performance liquid chromatography analysis were 3.7/12.3 and 5.7/18.9 μg L-1 , respectively. Recovery studies were also performed to show the applicability of the developed methods, and % recovery values ​​were found to be between 90-105% in tap water and 94-97% in wastewater for both analytes. The method was also successfully applied for the qualitative and quantitative determination of niacin and pyridoxine in drug samples. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  17. A high expression EGFR/cell membrane chromatography and online high performance liquid chromatography/mass spectrometry method for screening EGFR antagonists from Rhizoma Polygoni Cuspidati

    Directory of Open Access Journals (Sweden)

    Meng Sun

    2011-08-01

    Full Text Available The epidermal growth factor receptors (EGFRs in some tumor cells are significant targets for drug discovery. In this work, we have developed an EGFR cell membrane chromatography and online high performance liquid chromatography/mass spectrometry system for screening active component from Rhizoma Polygoni Cuspidati. As a result, resveratrol from Rhizoma Polygoni Cuspidati was found to be the active component acting on EGFR like gefitinib. There was a good relationship between their inhibiting effects on EGFR secretion and HEK293 EGFR cell growth in vitro. The EGFR/CMC-online-HPLC/MS system demonstrated fast and effective characteristics for screening leading compounds from traditional Chinese medicine.

  18. Separation and characterisation of beta2-microglobulin folding conformers by ion-exchange liquid chromatography and ion-exchange liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Bertoletti, Laura; Regazzoni, Luca; Aldini, Giancarlo; Colombo, Raffaella; Abballe, Franco; Caccialanza, Gabriele; De Lorenzi, Ersilia

    2013-04-10

    In this work we present for the first time the use of ion-exchange liquid chromatography to separate the native form and a partially structured intermediate of the folding of the amyloidogenic protein beta2-microglobulin. Using a strong anion-exchange column that accounts for the differences in charge exposure of the two conformers, a LC-UV method is initially optimised in terms of mobile phase pH, composition and temperature. The preferred mobile phase conditions that afford useful information were found to be 35 mM ammonium formate, pH 7.4 at 25°C. The dynamic equilibrium of the two species is demonstrated upon increasing the concentration of acetonitrile in the protein sample. Then, the chromatographic method is transferred to MS detection and the respective charge state distributions of the separated conformers are identified. The LC-MS results demonstrate that one of the conformers is partially unfolded, compared with the native and more compact species. The correspondence with previous results obtained in free solution by capillary electrophoresis suggest that strong ion exchange LC-MS does not alter beta2-microglobulin conformation and maintains the dynamic equilibrium already observed between the native protein and its folding intermediate. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. High-performance liquid chromatography/high-resolution mass spectrometry for the characterization of transformation products of ionic liquids.

    Science.gov (United States)

    Fabbri, Debora; Calza, Paola; Noè, Giorgio; Santoro, Valentina; Medana, Claudio

    2017-12-15

    Ionic liquids (ILs) are a subject of active research in the field of alternative solvents. We studied the behaviour of a piperidine IL, 1-butyl-1-methylpiperidinium tetrafluoroborate (BMPA), through the elucidation of its transformation products (TPs) in water. The transformation pathways of BMPA were investigated using high-performance liquid chromatography (HPLC) combined with a hybrid LTQ-Orbitrap instrument on the basis of mass defect filtering. TPs of BMPA were identified by fragmentation patterns and accurate mass measurements. The separation and identification of 32 TPs was achieved. BMPA can be oxidized at different positions in the alkyl chains. The ultimate products corresponds to N-methyl-piperidinium and some byproducts involving ring-opening. Tests of acute toxicity, evaluated with Vibrio Fischeri bacteria, show that BMPA transformation proceeds through the formation of slightly harmful compounds. Results showed that the main transformation pathways of BMPA were alkyl chain hydroxylation/shortening and de-alkylation, and that HPLC/LTQ-Orbitrap can serve as an important analytical platform to gather the unknown TPs of ILs. Copyright © 2017 John Wiley & Sons, Ltd.

  20. Determination of artificial sweeteners in sewage sludge samples using pressurised liquid extraction and liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Ordoñez, Edgar Y; Quintana, José Benito; Rodil, Rosario; Cela, Rafael

    2013-12-13

    An analytical method for the determination of six artificial sweeteners in sewage sludge has been developed. The procedure is based on pressurised liquid extraction (PLE) with water followed by solid-phase extraction (SPE) and subsequent liquid chromatography-tandem mass spectrometry analysis. After optimisation of the different PLE parameters, extraction with aqueous 500mM formate buffer (pH 3.5) at 80°C during a single static cycle of 21min proved to be best conditions. After a subsequent SPE, quantification limits, referred to dry weight (dw) of sewage sludge, ranged from 0.3ng/g for acesulfame (ACE) to 16ng/g for saccharin (SAC) and neohespiridine dihydrochalcone. The trueness, expressed as recovery, ranged between 72% and 105% and the precision, expressed as relative standard deviation, was lower than 16%. Moreover, the method proved its linearity up to the 2μg/g range. Finally, the described method was applied to the determination of the artificial sweeteners in primary and secondary sewage sludge from urban wastewater treatment plants. Four of the six studied artificial sweeteners (ACE, cyclamate, SAC and sucralose) were found in the samples at concentrations ranging from 17 to 628ng/g dw. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. Determination of thiobencarb in water samples by gas chromatography using a homogeneous liquid-liquid microextraction via flotation assistance procedure

    Directory of Open Access Journals (Sweden)

    H.A. Mashayekhi

    2013-09-01

    Full Text Available Homogeneous liquid-liquid microextraction via flotation assistance (HLLME-FA coupled with gas chromatography-flame ionization detection (GC-FID was applied for the extraction and determination of thiobencarb in water samples. In this study, a special extraction cell was designed to facilitate collection of the low-density solvent extraction. No centrifugation was required in this procedure. The water sample solution was added into the extraction cell which contained an appropriate mixture of toluene (as an extraction solvent and acetone (as a homogeneous solvent. By using air flotation, the organic solvent was collected at the conical part of the designed cell. The effect of the different parameters on the efficiency of extraction such as type and volume of extraction and homogeneous solvents, ionic strength and extraction time were studied and optimized. Under the optimal conditions, linearity of the method was in the range of 1.0-200 µg L-1. The relative standard deviations in the real samples varied from 7.8-11.7 % (n = 3. The proposed method was successfully applied to analysis of thiobencarb in the water samples and satisfactory results were obtained.DOI: http://dx.doi.org/10.4314/bcse.v27i3.4

  2. High-throughput screening for enzyme inhibitors using frontal affinity chromatography with liquid chromatography and mass spectrometry.

    Science.gov (United States)

    Ng, Ella S M; Yang, Feng; Kameyama, Akihiko; Palcic, Monica M; Hindsgaul, Ole; Schriemer, David C

    2005-10-01

    This work presents new frontal affinity chromatography (FAC) methodologies for high-throughput screening of compound libraries, designed to increase screening rates and improve sensitivity and ruggedness in performance. A FAC column constructed around the enzyme N-acetylglucosaminyltransferase V (GnT-V) was implemented in the identification of potential enzyme inhibitors from two libraries of trisaccharides. Effluent from the FAC column was fractionated, sequentially processed via LC/MS, and referenced to a similar analysis through a control FAC column lacking the enzyme. The resulting multidimensional data sets were compared across corresponding sample and control fractions to identify binders, in a semiautomated approach. A strong binder in the protonated form at m/z 795 was identified from the first library of 81 compounds, exhibiting an estimated Kd value of 0.3 microM. Other binders yielded Kd values ranging from 0.35 to 3.35 microM. To demonstrate the improvement in performance of this FAC-LC/MS approach over the conventional online FAC/MS approach, 15 compounds from this library were blended with a second library of 1000 synthetic trisaccharides and screened against GnT-V. All ligands in the 15-compound set were identified in this larger screen, and no ligands of greater affinity than compound 1 were found. Our results show that FAC-LC/MS is a reliable method for screening large compound libraries directly and useful for large-scale ligand discovery initiatives.

  3. EPA Method 8321B (SW-846): Solvent-Extractable Nonvolatile Compounds by High Performance Liquid Chromatography-Thermospray-Mass Spectrometry (HPLC-TS-MS) or Ultraviolet (UV) Detection

    Science.gov (United States)

    Method 8321B describes procedures for preparation and analysis of solid, aqueous liquid, drinking water and wipe samples using high performance liquid chromatography and mass spectrometry for extractable non-volatile compounds.

  4. High-performance liquid chromatography separation of unsaturated organic compounds by a monolithic silica column embedded with silver nanoparticles.

    Science.gov (United States)

    Zhu, Yang; Morisato, Kei; Hasegawa, George; Moitra, Nirmalya; Kiyomura, Tsutomu; Kurata, Hiroki; Kanamori, Kazuyoshi; Nakanishi, Kazuki

    2015-08-01

    The optimization of a porous structure to ensure good separation performances is always a significant issue in high-performance liquid chromatography column design. Recently we reported the homogeneous embedment of Ag nanoparticles in periodic mesoporous silica monolith and the application of such Ag nanoparticles embedded silica monolith for the high-performance liquid chromatography separation of polyaromatic hydrocarbons. However, the separation performance remains to be improved and the retention mechanism as compared with the Ag ion high-performance liquid chromatography technique still needs to be clarified. In this research, Ag nanoparticles were introduced into a macro/mesoporous silica monolith with optimized pore parameters for high-performance liquid chromatography separations. Baseline separation of benzene, naphthalene, anthracene, and pyrene was achieved with the theoretical plate number for analyte naphthalene as 36,000 m(-1). Its separation function was further extended to cis/trans isomers of aromatic compounds where cis/trans stilbenes were chosen as a benchmark. Good separation of cis/trans-stilbene with separation factor as 7 and theoretical plate number as 76,000 m(-1) for cis-stilbene was obtained. The trans isomer, however, is retained more strongly, which contradicts the long- established retention rule of Ag ion chromatography. Such behavior of Ag nanoparticles embedded in a silica column can be attributed to the differences in the molecular geometric configuration of cis/trans stilbenes. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Optimization of the Hewlett-Packard particle-beam liquid chromatography-mass spectrometry interface by statistical experimental design.

    Science.gov (United States)

    Huang, S K; Garza, N R

    1995-06-01

    Optimization of both sensitivity and ionization softness for the Hewlett-Packard particle-beam liquid chromatography-mass spectrometry interface has been achieved by using a statistical experimental design with response surface modeling. Conditions for both optimized sensitivity and ionization softness were found to occur at 55-lb/in.(2) nebulizer flow, 35°C desolvation chamber temperature with approximately 45% organic modifier in the presence of 0.02-F ammonium acetate and a liquid chromatography flow rate of 0.2 mL/min.

  6. Mutation analysis in {beta}{sub 2-}adrenergic receptor gene by denaturing high performance liquid chromatography (DHPLC)

    Energy Technology Data Exchange (ETDEWEB)

    Park, S.B.; Oh, C.H.; Kim, J.W.; Jang, W.C. [Dankook University, Cheonan (Korea)

    2002-06-01

    We analysed mutation of {beta}{sub 2-}adrenergic receptor gene that controls bronchial asthma by denaturing high performance liquid chromatography (DHPLC) according to ion-pair reversed-phase high performance liquid chromatography (IP-RP-HPLC). We extracted genomic DNA from 50 asthma patients, amplified DNA using PCR, and analysed PCR product by DHPLC. As a result, we obtained that mutation frequency was 15(30%) among 50 cases. Consequently DHPLC mutation detection was confirmed that the result of direct sequencing was coincide exactly. (author). 13 refs., 4 figs.

  7. A Simple and Universal Gel Permeation Chromatography Technique for Precise Molecular Weight Characterization of Well-Defined Poly(ionic liquid)s

    Energy Technology Data Exchange (ETDEWEB)

    He, Hongkun; Zhong, Mingjiang; Adzima, Brian; Luebke, David; Nulwala, Hunaid; Matyjaszewski, Krzysztof

    2013-03-20

    Poly(ionic liquid)s (PILs) are an important class of technologically relevant materials. However, characterization of well-defined polyionic materials remains a challenge. Herein, we have developed a simple and versatile gel permeation chromatography (GPC) methodology for molecular weight (MW) characterization of PILs with a variety of anions. PILs with narrow MW distributions were synthesized via atom transfer radical polymerization, and the MWs obtained from GPC were further confirmed via nuclear magnetic resonance end group analysis.

  8. Determination of capsaicinoids in Capsicum species using ultra performance liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Alothman, Zeid Abdullah; Wabaidur, Saikh Mohammad; Khan, Mohammad Rizwan; Abdel Ghafar, Ayman; Habila, Mohamed Abdelaty; Ahmed, Yacine Badjah Hadj

    2012-11-01

    In the present work, a rapid and sensitive ultra performance liquid chromatography-mass spectrometry method has been proposed for the analysis of capsaicinoids (nordihydrocapsaicin, capsaicin, dihydrocapsaicin, homocapsaicin, and homodihydrocapsaicin) present in different Capsicum samples. Extraction of capsaicinoids was carried out by liquid-liquid extraction using ethanol as an extracting solvent, while the chromatographic separation was achieved by reversed phase C(18) column with gradient mobile phase (solvent A: acetonitrile and solvent B: water with 0.1% formic acid). Under the optimum experimental conditions, the linear ranges were 0.5-50 μg/g with correlation coefficient (r(2) ) >0.999 for each capsaicinoids and detection limits were 0.15, 0.05, 0.06, 0.2, and 0.1 μg/g for nordihydrocapsaicin, capsaicin, dihydrocapsaicin, homocapsaicin, and homodihydrocapsaicin, respectively. Run-to-run and day-to-day precisions of the method with relative standard deviations capsaicinoids. The robustness of the method was determined by utilizing different injection volumes of the extracts. Furthermore, to validate the system robustness, a run of high number of capsaicinoids present in different varieties of Capsicum samples was performed in this study. All the capsaicinoids were separated in a time of less than 9 min by employing the proposed method. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Identification and quantification of dimethylamylamine in geranium by liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Li, J S; Chen, M; Li, Z C

    2012-01-01

    A sensitive and reliable method of liquid chromatography-electrospray ionization/tandem mass spectrometry (LC-ESI/MS/ MS) was developed and validated for determining 1,3-dimethylamylamine (1,3-DMAA) and 1,4-dimethylamylamine (1,4-DMAA) in geranium plants (Pelargonium graveolens). The sample was extracted with 0.5 M HCl and purified by liquid-liquid partition with hexane. The parameters for reverse-phase (C18) LC and positive ESI/MS/MS were optimized. The matrix effect, specificity, linearity, precision, accuracy and reproducibility of the method were determined and evaluated. The method was linear over a range of 0.10-10.00 ng/mL examined, with R(2) of 0.99 for both 1,3-DMAA and 1,4-DMAA. The recoveries from spiked concentrations between 5.00-40.00 ng/g were 85.1%-104.9% for 1,3-DMAA, with relative standard deviation (RSD) of 2.9%-11.0%, and 82.9%-101.8% for 1,4-DMAA, with RSD of 3.2%-11.7%. The instrument detection limit was 1-2 pg for both DMAAs. The quantification limit was estimated to be 1-2 ng/g for the plant sample. This method was successfully applied to the quantitative determination of 1,3- and 1,4-DMAA in both geranium plant and geranium oil.

  10. Determination of metformin in human plasma by high-performance liquid chromatography.

    Science.gov (United States)

    Amini, Hossein; Ahmadiani, Abolhassan; Gazerani, Parisa

    2005-09-25

    A simple, selective and sensitive high-performance liquid chromatographic method with spectrophotometric detection was developed for the determination of antihyperglycemic agent metformin in human plasma using a novel sample extraction procedure. Liquid-liquid extraction of metformin and ranitidine (as internal standard) from plasma samples was performed with 1-butanol/n-hexane (50:50, v/v) in alkaline condition followed by back-extraction into diluted acetic acid. Chromatography was carried out using a silica column (250 mmx4.6 mm, 5 microm) under isocratic elution with acetonitrile-40 mM aqueous sodium dihydrogen phosphate (25:75, v/v), pH 6. The limit of quantification (LOQ) was 15.6 ng/ml and the calibration curves were linear up to 2000 ng/ml. The mean absolute recoveries for metformin and internal standard using the present extraction procedure were 98 and 95%, respectively. The intra- and inter-day coefficient of variation and percent error values of the assay method were all less than 8.3%.

  11. Sensitive determination of clarithromycin in human plasma by high-performance liquid chromatography with spectrophotometric detection.

    Science.gov (United States)

    Amini, Hossein; Ahmadiani, Abolhassan

    2005-03-25

    A rapid, selective and sensitive high-performance liquid chromatographic method with spectrophotometric detection was developed for the determination of clarithromycin in human plasma. Liquid-liquid extraction of clarithromycin and norverapamil (as internal standard) from plasma samples was performed with n-hexane/1-butanol (98:2, v/v) in alkaline condition followed by back-extraction into diluted acetic acid. Chromatography was carried out using a CN column (250 mm x 4.6 mm, 5 microm) under isocratic elution with acetonitrile-50 mM aqueous sodium dihydrogen phosphate (32:68, v/v), pH 4.5. Detection was made at 205 nm and analyses were run at a flow-rate of 1.0 ml/min at 40 degrees C. The analysis time was less than 11 min. The method was specific and sensitive with a quantification limit of 31.25 ng/ml and a detection limit of 10 ng/ml in plasma. The mean absolute recovery of clarithromycin from plasma was 95.9%, while the intra- and inter-day coefficient of variation and percent error values of the assay method were all less than 9.5%. Linearity was assessed in the range of 31.25-2000 ng/ml in plasma with a correlation coefficient of greater than 0.999. The method was used to analyze several hundred human plasma samples for bioavailability studies.

  12. Scaling of Performance in Liquid Propellant Rocket Engine Combustion Devices

    Science.gov (United States)

    Hulka, James R.

    2008-01-01

    This paper discusses scaling of combustion and combustion performance in liquid propellant rocket engine combustion devices. In development of new combustors, comparisons are often made between predicted performance in a new combustor and measured performance in another combustor with different geometric and thermodynamic characteristics. Without careful interpretation of some key features, the comparison can be misinterpreted and erroneous information used in the design of the new device. This paper provides a review of this performance comparison, including a brief review of the initial liquid rocket scaling research conducted during the 1950s and 1960s, a review of the typical performance losses encountered and how they scale, a description of the typical scaling procedures used in development programs today, and finally a review of several historical development programs to see what insight they can bring to the questions at hand.

  13. Analysis of human plasma metabolites across different liquid chromatography/mass spectrometry platforms: Cross-platform transferable chemical signatures.

    Science.gov (United States)

    Telu, Kelly H; Yan, Xinjian; Wallace, William E; Stein, Stephen E; Simón-Manso, Yamil

    2016-03-15

    The metabolite profiling of a NIST plasma Standard Reference Material (SRM 1950) on different liquid chromatography/mass spectrometry (LC/MS) platforms showed significant differences. Although these findings suggest caution when interpreting metabolomics results, the degree of overlap of both profiles allowed us to use tandem mass spectral libraries of recurrent spectra to evaluate to what extent these results are transferable across platforms and to develop cross-platform chemical signatures. Non-targeted global metabolite profiles of SRM 1950 were obtained on different LC/MS platforms using reversed-phase chromatography and different chromatographic scales (conventional HPLC, UHPLC and nanoLC). The data processing and the metabolite differential analysis were carried out using publically available (XCMS), proprietary (Mass Profiler Professional) and in-house software (NIST pipeline). Repeatability and intermediate precision showed that the non-targeted SRM 1950 profiling was highly reproducible when working on the same platform (relative standard deviation (RSD) platforms or even using different chromatographic scales (conventional HPLC, UHPLC and nanoLC) on the same platform. A substantial degree of overlap (common molecular features) was also found. A procedure to generate consistent chemical signatures using tandem mass spectral libraries of recurrent spectra is proposed. Different platforms rendered significantly different metabolite profiles, but the results were highly reproducible when working within one platform. Tandem mass spectral libraries of recurrent spectra are proposed to evaluate the degree of transferability of chemical signatures generated on different platforms. Chemical signatures based on our procedure are most likely cross-platform transferable. Published in 2016. This article is a U.S. Government work and is in the public domain in the USA.

  14. A New Liquid Chromatography-Tandem Mass Spectrometry Method for Determination of Bisoprolol in Human Plasma Samples

    Directory of Open Access Journals (Sweden)

    Gabriela Peste

    2009-01-01

    Full Text Available Liquid chromatography (LC coupled with mass spectrometry (MS detection is one of the most powerful analytical tools for organic compound analysis. The advantages of using LC/MS methods over HPLC methods include: selectivity, chromatographic integrity, peak assignment, structural information, and rapid method development. In this paper, a new liquid chromatography-tandem mass spectrometry (LC-MS/MS method has been developed and validated for the determination of bisoprolol in human plasma samples, using metoprolol as internal standard and liquid-liquid extraction procedure. The assay has proven to be sensitive, specific and reproducible, suitable to determine the bisoprolol concentration, following a single oral administration of a 10 mg bisoprolol tablet in 22 healthy volunteers, in the bioequivalence study of Bisoprolol 10 mg coated tablets, produced by Antibiotice S.A. versus Concor 10 mg, produced by Merck.

  15. Two dimensional assisted liquid chromatography - a chemometric approach to improve accuracy and precision of quantitation in liquid chromatography using 2D separation, dual detectors, and multivariate curve resolution.

    Science.gov (United States)

    Cook, Daniel W; Rutan, Sarah C; Stoll, Dwight R; Carr, Peter W

    2015-02-15

    Comprehensive two-dimensional liquid chromatography (LC×LC) is rapidly evolving as the preferred method for the analysis of complex biological samples owing to its much greater resolving power compared to conventional one-dimensional (1D-LC). While its enhanced resolving power makes this method appealing, it has been shown that the precision of quantitation in LC×LC is generally not as good as that obtained with 1D-LC. The poorer quantitative performance of LC×LC is due to several factors including but not limited to the undersampling of the first dimension and the dilution of analytes during transit from the first dimension ((1)D) column to the second dimension ((2)D) column, and the larger relative background signals. A new strategy, 2D assisted liquid chromatography (2DALC), is presented here. 2DALC makes use of a diode array detector placed at the end of each column, producing both multivariate (1)D and two-dimensional (2D) chromatograms. The increased resolution of the analytes provided by the addition of a second dimension of separation enables the determination of analyte absorbance spectra from the (2)D detector signal that are relatively pure and can be used to initiate the treatment of data from the first dimension detector using multivariate curve resolution-alternating least squares (MCR-ALS). In this way, the approach leverages the strengths of both separation methods in a single analysis: the (2)D detector data is used to provide relatively pure analyte spectra to the MCR-ALS algorithm, and the final quantitative results are obtained from the resolved (1)D chromatograms, which has a much higher sampling rate and lower background signal than obtained in conventional single detector LC×LC, to obtain accurate and precise quantitative results. It is shown that 2DALC is superior to both single detector selective or comprehensive LC×LC and 1D-LC for quantitation of compounds that appear as severely overlapped peaks in the (1)D chromatogram - this is

  16. Analysis of eight capsaicinoids in peppers and pepper-containing foods by high-performance liquid chromatography and liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Kozukue, Nobuyuki; Han, Jae-Sook; Kozukue, Etsuko; Lee, Sin-Jung; Kim, Joung-Ae; Lee, Kap-Rang; Levin, Carol E; Friedman, Mendel

    2005-11-16

    Diverse procedures have been reported for the isolation and analysis of secondary metabolites called capsaicinoids, pungent compounds in the fruit of the Capsicum (Solanaceae) plant. To further improve the usefulness of high-performance liquid chromatography (HPLC), studies were carried out on the analysis of extracts containing up to eight of the following capsaicinoids: capsaicin, dihydrocapsaicin, homocapsaicin-I, homocapsaicin-II, homodihydrocapsaicin-I, homodihydrocapsaicin-II, nonivamide, and nordihydrocapsaicin. HPLC was optimized by defining effects on retention times of (a) the composition of the mobile phase (acetonitrile/0.5% formic acid in H2O), (b) the length of the Inertsil column, and (c) the capacity values (k) of the column packing. Identification was based on retention times and mass spectra of individual peaks. Quantification was based on the UV response at 280 nm in HPLC and recoveries from spiked samples. The method (limit of detection of approximately 15-30 ng) was successfully used to quantify capsaicinoid levels of parts of the pepper fruit (pericarp, placenta, seeds, and in the top, middle, and base parts of whole peppers) in 17 species of peppers and in 23 pepper-containing foods. The results demonstrate the usefulness of the method for the analysis of capsaicinoids ranging from approximately 0.5 to 3600 microg of capsaicin equiv/g of product. The water content of 12 fresh peppers ranged from 80.8 to 92.7%. The described freeze-drying, extraction, and analysis methods should be useful for assessing the distribution of capsaicinoids in the foods and in defining the roles of these biologically active compounds in the plant, the diet, and medicine.

  17. Liquid crystals in micron-scale droplets, shells and fibers

    Science.gov (United States)

    Urbanski, Martin; Reyes, Catherine G.; Noh, JungHyun; Sharma, Anshul; Geng, Yong; Subba Rao Jampani, Venkata; Lagerwall, Jan P. F.

    2017-04-01

    The extraordinary responsiveness and large diversity of self-assembled structures of liquid crystals are well documented and they have been extensively used in devices like displays. For long, this application route strongly influenced academic research, which frequently focused on the performance of liquid crystals in display-like geometries, typically between flat, rigid substrates of glass or similar solids. Today a new trend is clearly visible, where liquid crystals confined within curved, often soft and flexible, interfaces are in focus. Innovation in microfluidic technology has opened for high-throughput production of liquid crystal droplets or shells with exquisite monodispersity, and modern characterization methods allow detailed analysis of complex director arrangements. The introduction of electrospinning in liquid crystal research has enabled encapsulation in optically transparent polymeric cylinders with very small radius, allowing studies of confinement effects that were not easily accessible before. It also opened the prospect of functionalizing textile fibers with liquid crystals in the core, triggering activities that target wearable devices with true textile form factor for seamless integration in clothing. Together, these developments have brought issues center stage that might previously have been considered esoteric, like the interaction of topological defects on spherical surfaces, saddle-splay curvature-induced spontaneous chiral symmetry breaking, or the non-trivial shape changes of curved liquid crystal elastomers with non-uniform director fields that undergo a phase transition to an isotropic state. The new research thrusts are motivated equally by the intriguing soft matter physics showcased by liquid crystals in these unconventional geometries, and by the many novel application opportunities that arise when we can reproducibly manufacture these systems on a commercial scale. This review attempts to summarize the current understanding of

  18. Multiclass mycotoxin analysis in edible oils using a simple solvent extraction method and liquid chromatography with tandem mass spectrometry.

    Science.gov (United States)

    Eom, Taeyong; Cho, Hyun-Deok; Kim, Junghyun; Park, Mihee; An, Jinyoung; Kim, Moosung; Kim, Sheen-Hee; Han, Sang Beom

    2017-11-01

    A simple and rapid method for the simultaneous determination of 11 mycotoxins - aflatoxins B1, B2, G1 and G2; fumonisins B1, B2 and B3; ochratoxin A; zearalenone; deoxynivalenol; and T-2 toxin - in edible oils was established using liquid chromatography tandem mass spectrometry (LC-MS/MS). In this study, QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe), QuEChERS with dispersive liquid-liquid microextraction, and solvent extraction were examined for sample preparation. Among these methods, solvent extraction with a mixture of formic acid/acetonitrile (5/95, v/v) successfully extracted all target mycotoxins. Subsequently, a defatting process using n-hexane was employed to remove the fats present in the edible oil samples. Mass spectrometry was carried out using electrospray ionisation in polarity switching mode with multiple reaction monitoring. The developed LC-MS/MS method was validated by assessing the specificity, linearity, recovery, limit of quantification (LOQ), accuracy and precision with reference to Commission Regulation (EC) 401/2006. Mycotoxin recoveries of 51.6-82.8% were achieved in addition to LOQs ranging from 0.025 ng/g to 1 ng/g. The edible oils proved to be relatively uncomplicated matrices and the developed method was applied to 9 edible oil samples, including soybean oil, corn oil and rice bran oil, to evaluate potential mycotoxin contamination. The levels of detection were significantly lower than the international regulatory standards. Therefore, we expect that our developed method, based on simple, two-step sample preparation process, will be suitable for the large-scale screening of mycotoxin contamination in edible oils.

  19. Development of high performance liquid chromatography method for miconazole analysis in powder sample

    Science.gov (United States)

    Hermawan, D.; Suwandri; Sulaeman, U.; Istiqomah, A.; Aboul-Enein, H. Y.

    2017-02-01

    A simple high performance liquid chromatography (HPLC) method has been developed in this study for the analysis of miconazole, an antifungal drug, in powder sample. The optimized HPLC system using C8 column was achieved using mobile phase composition containing methanol:water (85:15, v/v), a flow rate of 0.8 mL/min, and UV detection at 220 nm. The calibration graph was linear in the range from 10 to 50 mg/L with r 2 of 0.9983. The limit of detection (LOD) and limit of quantitation (LOQ) obtained were 2.24 mg/L and 7.47 mg/L, respectively. The present HPLC method is applicable for the determination of miconazole in the powder sample with a recovery of 101.28 % (RSD = 0.96%, n = 3). The developed HPLC method provides short analysis time, high reproducibility and high sensitivity.

  20. ANALISIS RESIDU KLORPIRIFOS DALAM SAYUR-SAYURAN DENGAN TEKNIK HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC

    Directory of Open Access Journals (Sweden)

    Aman Sentosa Panggabean

    2016-06-01

    Full Text Available The research about analysis of chlorpyrifos residue in vegetables by using High Performance Liquid Chromatography (HPLC technique has been done. To obtain the optimal measurement results, the measurement performed several important parameters in the chromatographic system was composition of mobile phase, volume injection sample, flow rate and pH eluent. Optimum measurement conditions obtained was mobile phase composition (water : methanol with 70 : 30, volume injection sample are 5 mL, flow rate are 0.5mL/menit and pH eluent are 7. The analytical performance that obtained is good showed with the reproducibility value as percentage coefficient variance (% CV was 0.0664%, limit of detection (LOD was 0.44 ppm, with a recovery percentage of > 95%. The results obtained showed the HPLC technique can be used for the routine analysis in the determination of chlorpyrifos for the vegetable samples. Keywords: Chlorpyrifos, Vegetables, HPLC.

  1. Preparation and evaluation of pillararene bonded silica gel stationary phases for high performance liquid chromatography.

    Science.gov (United States)

    Zhao, Wenjie; Chu, Jianxiang; Xie, Fuwei; Duan, Qunpeng; He, Lijun; Zhang, Shusheng

    2017-02-17

    Pillararene bonded stationary phases for high performance liquid chromatography were prepared using 3-aminopropyltriethoxysilane as coupling reagent. The structure of the new materials was characterized by infrared spectroscopy, elemental analysis and thermogravimetric analysis. The chromatographic performance and retention mechanism of the new stationary phases were evaluated in reversed-phase mode compared with C18 using different solute probes including Tanaka test solutes, polycyclic aromatic hydrocarbons, phenols and aromatic positional isomers. The new stationary phases could provide various interactions for different solutes, such as hydrophobic, hydrogen bonding, π-π and inclusion interactions. The synergistic effects resulting from aromatic rings, oxygen atoms, alkyl linkers and cavities in the new host molecules improved the separation selectivity by multiple retention mechanisms. Such hybrid stationary phases can provide more versatility and have great potential for the analysis of complex samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Analysis of sulfonated compounds by ion-exchange high-performance liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Socher, G; Nussbaum, R; Rissler, K; Lankmayr, E

    2001-03-30

    Ion-exchange high-performance liquid chromatography (HPIEC)-mass spectrometry (MS) was used for the analysis of different sulfonated compounds. HPIEC was performed on an aminopropyl column applying a gradient with increasing concentration of a buffer consisting of ammonium acetate-acetic acid and acetonitrile as the organic modifier. HPIEC is well suited to highly efficient separation of sulfonated compounds and furthermore, due to the volatility of ammonium acetate, the method is also appropriate for LC-MS coupling by the means of either atmospheric pressure chemical ionization or electrospray ionization. The applicability range of HPIEC-MS is demonstrated on the basis of a complex mixture of model substances consisting of sulfonated aromatics and textile dyes largely differing from each other in their structural properties.

  3. Quantitative determination of capsaicinoids by liquid chromatography-electrospray mass spectrometry.

    Science.gov (United States)

    Thompson, Robert Q; Phinney, Karen W; Welch, Michael J; White, Edward

    2005-04-01

    Eight naturally occurring capsaicinoids have been determined in Capsicum by use of high-purity standards, with norcapsaicin as an internal standard. The solid standards were rigorously checked for purity. The sensitivity of electrospray ionization (ESI), atmospheric-pressure chemical ionization (APCI), and coordination ion-spray (CIS; with silver) toward the capsaicinoids were measured and compared. The highest sensitivity was found for positive-ion ESI. Method validation of the liquid chromatography-ESI-mass spectrometry (LC-ESI-MS) determination is reported, including tests for repeatability (4%), detection limit (5 pg injected), linear range (20-6 ng injected), quantitation (excellent linearity; capsaicinoids in a commercial plant extract and in chili pepper fruits were quantified.

  4. Core-Shell Columns in High-Performance Liquid Chromatography: Food Analysis Applications

    Directory of Open Access Journals (Sweden)

    Raffaella Preti

    2016-01-01

    Full Text Available The increased separation efficiency provided by the new technology of column packed with core-shell particles in high-performance liquid chromatography (HPLC has resulted in their widespread diffusion in several analytical fields: from pharmaceutical, biological, environmental, and toxicological. The present paper presents their most recent applications in food analysis. Their use has proved to be particularly advantageous for the determination of compounds at trace levels or when a large amount of samples must be analyzed fast using reliable and solvent-saving apparatus. The literature hereby described shows how the outstanding performances provided by core-shell particles column on a traditional HPLC instruments are comparable to those obtained with a costly UHPLC instrumentation, making this novel column a promising key tool in food analysis.

  5. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR DETERMINATION OF NIFEDIPINE IN HUMAN PLASMA

    Directory of Open Access Journals (Sweden)

    MOHAMMAD ABDOLLAHI

    1999-10-01

    Full Text Available A relatively simple normal phase high performance liquid chromatography (HPLC method was modified for determination of nifedipine in human plasma. The method is based on ultraviolet detection at 235 nm and acidic plasma extraction by a mixture of dichioromethane (30% and n-hexane (70% using nimodipine as an internal standard. The system was stabilized with the use of n-hexane (80%, chloroform (17% and methanol (3% as mobile phase. The assay was linear up to at least 120 ng/ml of nifedipine in plasma. The limit of reliable determination was at least 3 ng/ml plasma. The reproducibility of the method was satisfactory. The procedure can be used effectively to quantitate nifedipine in the human plasma.

  6. [Determination of polysorbates in foods by solid-phase extraction and high-performance liquid chromatography].

    Science.gov (United States)

    Nomura, Chie; Kitagawa, Mikiya; Yoshida, Masaharu; Tanaka, Yukio

    2007-06-01

    A simple and rapid method using refractive index high-performance liquid chromatography (RI-HPLC) was developed for the determination of polysorbates (PS) in processed foods. PS were extracted with ethyl acetate containing 5% methanol. The extract was cleaned up on a multimode cartridge (300 mg) and an Alumina-N cartridge (500 mg) to remove fats and food color. HPLC separation was performed on a C18 column (4.6 i.d. x 150 mm) with methanol as the mobile phase. The recoveries of PS80 from nine kinds of foods fortified at the levels 1-5 g/kg were 80-99%. The limit of quantitation for PS80 in foods was 0.10 g/kg. The proposed method was applied to Worcestershire sauce that was PS-positive by TLC, and PS was confirmed to be present as PS80 at the concentration of 0.13 g/kg.

  7. Characterization of nine polyphenols in fruits of Malus pumila Mill by high-performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Lu Bai

    2016-04-01

    Full Text Available Polyphenols are important bioactive substances in apple. To explore the profiles of the nine representative polyphenols in this fruit, a high-performance liquid chromatography method has been established and validated. The validated method was successfully applied for the simultaneous characterization and quantification of these nine apple polyphenols in 11 apple extracts, which were obtained from six cultivars from Shaanxi Province, China. The results showed that only abscission of the Fuji apple sample was rich in the nine apple polyphenols, and the polyphenol contents of other samples varied. Although all the samples were collected in the same region, the contents of nine polyphenols were different. The proposed method could serve as a prerequisite for quality control of Malus products.

  8. Characterization of nine polyphenols in fruits of Malus pumila Mill by high-performance liquid chromatography.

    Science.gov (United States)

    Bai, Lu; Guo, Sen; Liu, Qingchao; Cui, Xueqin; Zhang, Xinxin; Zhang, Li; Yang, Xinwen; Hou, Manwei; Ho, Chi-Tang; Bai, Naisheng

    2016-04-01

    Polyphenols are important bioactive substances in apple. To explore the profiles of the nine representative polyphenols in this fruit, a high-performance liquid chromatography method has been established and validated. The validated method was successfully applied for the simultaneous characterization and quantification of these nine apple polyphenols in 11 apple extracts, which were obtained from six cultivars from Shaanxi Province, China. The results showed that only abscission of the Fuji apple sample was rich in the nine apple polyphenols, and the polyphenol contents of other samples varied. Although all the samples were collected in the same region, the contents of nine polyphenols were different. The proposed method could serve as a prerequisite for quality control of Malus products. Copyright © 2015. Published by Elsevier B.V.

  9. Analysis of bromate in drinking water using liquid chromatography-tandem mass spectrometry without sample pretreatment.

    Science.gov (United States)

    Kosaka, Koji; Asami, Mari; Takei, Kanako; Akiba, Michihiro

    2011-01-01

    An analytical method for determining bromate in drinking water was developed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The (18)O-enriched bromate was used as an internal standard. The limit of quantification (LOQ) of bromate was 0.2 µg/L. The peak of bromate was separated from those of coexisting ions (i.e., chloride, nitrate and sulfate). The relative and absolute recoveries of bromate in two drinking water samples and in a synthesized ion solution (100 mg/L chloride, 10 mg N/L nitrate, and 100 mg/L sulfate) were 99-105 and 94-105%, respectively. Bromate concentrations in 11 drinking water samples determined by LC-MS/MS were water without sample pretreatment.

  10. Monitoring carboxylic acid formation in engine oils by liquid chromatography with fluorescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Lewis, Simon W.; Worsfold, Paul J. (Dept. of Environmental Sciences, Univ. of Plymouth (United Kingdom)); McKerrell, Euan H. (Shell Research Ltd., Thornton Research Centre, Chester (United Kingdom))

    1994-04-21

    Straight chain aliphatic carboxylic acids (C[sub 6]-C[sub 22]) were selectively derivatised in oxidised engine oils with the fluorescent label 9-anthracenemethanol after dialysis of the oils to remove polymeric additives, organometallic oxidation products and solid debris. The ester derivatives were separated by reversed-phase liquid chromatography and quantified using fluorescence detection ([lambda][sub ex]=251 nm; [lambda][sub em]=412 nm). Calibrations over the range 1-4 mg ml[sup -1] were linear (0.9987[<=]r[sup 2][<=]1.0000). The limit of detection (S/N=3) for octadecanoic acid was 85 [mu]g ml[sup -1] in n-heptane (4.3 ng on-column). Results showed that carboxylic acids are formed during the oxidation of oils in car engines and that their concentrations are directly related to the degree of oil degradation

  11. Use of high-performance liquid chromatography-diode array detection in forensic toxicology.

    Science.gov (United States)

    Koves, E M

    1995-02-10

    A comprehensive approach to the analysis for many drugs in postmortem blood and biological fluids using high-performance liquid chromatography and diode array detection has been developed. To reduce the likelihood of co-eluting interference components of postmortem blood or other drugs, selective back-extraction was also used to screen and quantitate drugs in blood and biofluids. An isocratic mobile phase (acetonitrile, phosphoric acid and triethylamine buffer, pH 3.4) was developed and found stable, reliable and convenient for general drug screening and quantitation. A library of drug spectra in the ultraviolet wavelength range (210-367 nm) was established for 272 drugs on two reversed-phase columns: Supelcosil (biphenyl) and LiChrospher RP-8. The application of several methods to whole blood, the analysis of complex cases and the use of multicomponent analysis for qualitative and quantitative analysis is discussed.

  12. Determination of total L-Ascorbic Acid by high performance liquid chromatography in human plasma

    Directory of Open Access Journals (Sweden)

    Oveisi MR

    2001-07-01

    Full Text Available The total vitamin C content in human plasma is widely accepted as an indicator of the tissue status of vitamin C. A liquid chromatography method with ultraviolet detector (264 nm for measuring ascorbic acid in human plasma was developed. A C18 reversed-phase column and cetrimide as an ion-pairing agent was employed. Ascorbic acid (AA was measured after reducing L-dehydroascorbic acid to L-ascorbic acid with dithiothreitol. The stability of the ascorbic acid in plasma, metaphosphoric acid and trichloroacetic acid was also evaluated. The analytical parameters, including linearity (1-60 µg/ml, accuracy (98.98%, repeatability (2.8% and reproducibility (7.2%, showed that the method is reliable for measuring the total vitamin C content in plasma.

  13. SIMULTANEOUS DETERMINATION OF PARACETAMOL AND IBUPROFENE MIXTURES BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    Sophi Damayanti

    2010-06-01

    Full Text Available Analytical method for the determination of paracetamol and ibuprofene mixtures has been developed by High Performance Liquid Chromatography using C-18 column and acetinitrile - phosphate buffer pH = 4.5 (75:25 containing 0.075% sodium hexanesulfunate as a mobile phase. The detector was set at 215 nm. Using such conditions, retention time for paracetamol and ibuprofen was 4.89 and 7.11 min, respectively. The recovery for paracetamol and ibuprofen was found to be 101.07± 0.73% and 102.02 ± 1.58%, respectively. The detector limits of the method was 1.30 and 1.60 μg/mL with the relative standard deviation (RSD 0.74 and 1.52% for paracetamol and ibuprofen, respectively.   Keywords: paracetamol, ibuprofen, multi-component, validation, HPLC.

  14. Quantitative analysis of urine vapor and breath by gas-liquid partition chromatography.

    Science.gov (United States)

    Pauling, L; Robinson, A B; Teranishi, R; Cary, P

    1971-10-01

    When a human being is placed for several days on a completely defined diet, consisting almost entirely of small molecules that are absorbed from the stomach into the blood, intestinal flora disappear because of lack of nutrition. By this technique, the composition of body fluids can be made constant (standard deviation about 10%) after a few days, permitting significant quantitative analyses to be performed. A method of temperature-programmed gas-liquid partition chromatography has been developed for this purpose. It permits the quantitative determination of about 250 substances in a sample of breath, and of about 280 substances in a sample of urine vapor. The technique should be useful in the application of the principles of orthomolecular medicine.

  15. Glucose and glycerol concentrations and their tracer enrichment measurements using liquid chromatography tandem mass spectrometry

    DEFF Research Database (Denmark)

    Bornø, Andreas; Foged, Lene; van Hall, Gerrit

    2014-01-01

    in connection to in vivo metabolic studies investigating glucose turnover and lipolytic rate. Moreover, in order to keep up with this new fast analysis, simple derivatization procedures have been developed. Prior to analysis, glucose and glycerol were derivatized using benzoyl chloride in order to form......The present study describes a new liquid chromatography tandem mass spectrometry method for high-throughput quantification of glucose and glycerol in human plasma using stable isotopically labeled internal standards and is suitable for simultaneous measurements of glucose and glycerol enrichments...... benzoylated derivatives via new simplified fast procedures. For glucose, two internal standards were evaluated, [U-(13) C(6)]glucose and [U-(13) C(6), D(7)]glucose, and for glycerol, [U-(13) C(3), D(8)]glycerol was used. The method was validated by means of calibration curves, quality control samples...

  16. High-performance liquid chromatography for determination of α-tocopherol in vegetables

    Directory of Open Access Journals (Sweden)

    Marcin Horbowicz

    2013-12-01

    Full Text Available A simple method for the determination of α-tocopherol in vegetables is described. The procedure consists of the following steps: saponification, extraction, silica-column clean-up, and high-performance liquid chromatography. Elution time for D, L-α-tocopherol was 9.0 min using a Zorbax Sil (250 x 4.6 mm column and an isocratic mobile phase of hexane-methanol (99.3 + 0.7, with a flow rate of 1 ml/min, and detection at 292 nm using a variable UV detector. The average recovery of α-tocopherol was 91.2%, and the minimum detectable amount was 0.1 mg/100 g of fresh vegetable tissue. This method is comparable to gas-chromatographic determination of α-tocopherol, but has fewer analytical steps and gives more reproducible results.

  17. An entropy-based method for noise reduction of liquid chromatography-mass spectrometry data.

    Science.gov (United States)

    Li, Yunfei; Qu, Haibin; Cheng, Yiyu

    2008-03-31

    Entropy-based methods have been extensively used to measure the uncertainty information in a variety of fields. In this article, a novel information theory-based method for reducing noise of liquid chromatography-mass spectrometry (LC/MS) data was developed. The uncertainty existed in the LC/MS chromatograms was captured and evaluated by information entropy. By comparing the information entropy computationally derived from mass chromatograms, the good quality chromatograms and the noisy chromatograms can be distinguished. The proposed method was applied in processing LC/MS data of "Jing-Zhi-Guan-Xin" troche which is a well-known preparation of traditional Chinese medicine (TCM). The obtained result indicated that this method is beneficial to reduce noise of LC/MS data of complicated chemical samples, such as TCM.

  18. Determination of {sup 90}SR in food using extraction chromatography and LSC - Liquid Scintillation Counting

    Energy Technology Data Exchange (ETDEWEB)

    Temba, Eliane S.C.; Amaral, Angela M.; Reis Junior, Aluisio S.; Monteiro, Roberto P.G., E-mail: esct@cdtn.br [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG/SERTA), Belo Horizonte, MG (Brazil). Servico de Reator e Tecnicas Analiticas

    2013-07-01

    A methodology for the determination of {sup 90}Sr in food is described. The procedure involved a preliminary freeze-drying of the samples followed by dry-ashing and sample digestion. The separation procedure was carried out using extraction chromatography with Sr Resin, from Eichrom, and {sup 90}Sr was measured by liquid scintillation counting (LSC). A certified reference material, IAEA-375, was analyzed in order to evaluate the reliability of the method, and the results showed good agreement between the measured and certified values. The chemical yield was above 90% and the typical counting efficiency was 82%. The calculated limit of detection was 4.8 x 10{sup -3} Bq g{sup -1}. (author)

  19. Detection of cheese whey and caseinomacropeptide in fermented milk beverages using high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    E.H.P. Andrade

    2014-06-01

    Full Text Available Cheese whey level and caseinomacropeptide (CMP index of fermented milk beverages added with four levels of cheese whey (0, 10, 20, and 40% and stored at 8-10oC for 0, 7, 14 and 21 days were determined by high performance liquid chromatography-gel filtration (HPLC-GF. Additionally, the interference of the starter culture and the storage time on the detection of cheese whey and CMP were investigated. Refrigerated storage up to 21 days did not affect (P>0.05 cheese whey and CMP amounts in milk (0% of cheese whey and in fermented milk beverages added with 10 and 20% of cheese whey (P>0.05. However, cheese whey and CMP amounts were higher than expected (P<0.05 in fermented milk beverages added with 40% of cheese whey and stored for 21 days.

  20. Determination of dapsone in meat and milk by liquid chromatography tandem mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Hadjigeorgiou, M. [State General Laboratory of Cyprus, Veterinary Drug Residues Lab, Kimonos 44, 1451 Nicosia (Cyprus)], E-mail: mhadjigeorgiou@sgl.moh.gov.cy; Papachrysostomou, Ch.; Theodorou, Z.; Kanari, P.; Constantinou, S. [State General Laboratory of Cyprus, Veterinary Drug Residues Lab, Kimonos 44, 1451 Nicosia (Cyprus)

    2009-04-01

    Within the EU the use of dapsone (4,4-diaminodiphenylsulfone) is prohibited in food-producing animals and consequently it's included in the Annex IV of the Directive 90/2377/EC. A quantitative confirmatory method has been developed and validated according to the criteria defined in the Commission Decision 2002/657/EC, for the determination of dapsone in meat and milk. Samples, after homogenization in alkaline conditions and organic solvent extraction, were purified on silica gel solid phase extraction cartridges. The eluate was evaporated and redissolved in mobile phase and was analysed by liquid chromatography tandem mass spectrometry (LC-MS/MS) in positive electrospray ionisation (ESI) using deuterium labelled Sulphadimidine-d7 as internal standard. The calculated value for, decision limit, CC{alpha} is 0.12 {mu}g kg{sup -1}, and the detection capability; CC{beta} value is 0.16 {mu}g kg{sup -1}.

  1. Analysis of lipophilic pigments from a phototrophic microbial mat community by high performance liquid chromatography

    Science.gov (United States)

    Palmisano, A. C.; Cronin, S. E.; Des Marais, D. J.

    1988-01-01

    As assay for lipophilic pigments in phototrophic microbial mat communities using reverse phase-high performance liquid chromatography was developed which allows the separation of 15 carotenoids and chloropigments in a single 30 min program. Lipophilic pigments in a laminated mat from a commercial salina near Laguna Guerrero Negro, Baja California Sur, Mexico reflected their source organisms. Myxoxanthophyll, echinenone, canthaxanthin, and zeaxanthin were derived from cyanobacteria; chlorophyll c, and fucoxanthin from diatoms; chlorophyll a from cyanobacteria and diatoms; bacteriochlorophylls a and c, bacteriophaeophytin a, and gamma-carotene from Chloroflexus spp.; and beta-carotene from a variety of phototrophs. Sensitivity of detection was 0.6-6.1 ng for carotenoids and 1.7-12 ng for most chloropigments. This assay represents a significant improvement over previous analyses of lipophilic pigments in microbial mats and promises to have a wider application to other types of phototrophic communities.

  2. Analytical procedure for the determination of tetracyclines in medicated feedingstuffs by liquid chromatography-mass spectrometry

    Directory of Open Access Journals (Sweden)

    Patyra Ewelina

    2016-03-01

    Full Text Available Introduction: The article presents a rapid and simple analytical procedure for determination of four tetracyclines (oxytetracycline, tetracycline, chlortetracycline, and doxycycline in animal medicated feedingstuffs. Material and Methods: Two-gramme samples were extracted by a Na2EDTA-McIlvaine buffer (pH 4/methanol mixtures (40/60, v/v. The determination was achieved by liquid chromatography using a Zorbax Eclipse XDB C18 analytical column with mass spectrometer detection (LC-MS. Results: Recoveries of the antibiotics from spiked feed samples ranged from 78.2% to 113.5%. The LOD and LOQ for tetracyclines in feeds ranged from 2.8 to 4.2 and from 4.3 to 5.7 mg/kg, respectively. Conclusion: The method was successfully validated and proved to be efficient, precise, and useful for quantification of tetracyclines in medicated feedingstuffs.

  3. Uncovering biologically significant lipid isomers with liquid chromatography, ion mobility spectrometry and mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Kyle, Jennifer E.; Zhang, Xing; Weitz, Karl K.; Monroe, Matthew E.; Ibrahim, Yehia M.; Moore, Ronald J.; Cha, Jeeyeon; Sun, Xiaofei; Lovelace, Erica S.; Wagoner, Jessica; Polyak, Steve; Metz, Thomas O.; Dey, Sudhansu K.; Smith, Richard D.; Burnum-Johnson, Kristin E.; Baker, Erin Shammel

    2016-01-01

    Understanding how biological molecules are generated, metabolized and eliminated in living systems is important for interpreting processes such as immune response and disease pathology. While genomic and proteomic studies have provided vast amounts of information over the last several decades, interest in lipidomics has also grown due to improved analytical technologies revealing altered lipid metabolism in type 2 diabetes, cancer, and lipid storage disease. Liquid chromatography and mass spectrometry (LC-MS) measurements are currently the dominant approach for characterizing the lipidome by providing detailed information on the spatial and temporal composition of lipids. However, interpreting lipids’ biological roles is challenging due to the existence of numerous structural and stereoisomers (i.e. distinct acyl chain and double-bond positions), which are unresolvable using present LC-MS approaches. Here we show that combining structurally-based ion mobility spectrometry (IMS) with LC-MS measurements distinguishes lipid isomers and allows insight into biological and disease processes.

  4. [Determination of pesticides in Chinese dumplings using liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Okamoto, You; Takatori, Satoshi; Kitagawa, Yoko; Okihashi, Masahiro; Fukui, Naoki; Murata, Hiroshi; Sumimoto, Tatsuo; Tanaka, Yukio; Obana, Hirotaka

    2009-02-01

    A rapid and easy multiresidue method for determination of pesticide residues in Chinese dumplings using liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed. Pesticide residues were extracted with ethyl acetate in the presence of anhydrous magnesium sulfate in a disposable tube using a homogenizer. The extract was concentrated and reconstituted in hexane, followed by acetonitrile-hexane partition to remove lipids. The acetonitrile layer was purified with a double-layered cartridge column (graphite carbon black/primary secondary amine silica gel). After removal of the solvent, the extract was resolved in methanol/water and analyzed with LC-MS/MS. Recovery tests of 99 pesticide residues from Chinese dumpling were performed at 20 and 100 ng/g, and 72 pesticides exhibited acceptable recoveries (70-120%) with low relative standard deviations (pesticide residues in the Chinese dumplings.

  5. Liquid chromatography/coordination ion spray-mass spectrometry for the analysis of rubber vulcanization products.

    Science.gov (United States)

    Hayen, Heiko; Alvarez-Grima, M Montserrat; Debnath, Subhas C; Noordermeer, Jacques W M; Karst, Uwe

    2004-02-15

    Liquid chromatography/coordination ion spray-mass spectrometry has been used for the identification of reaction products in a model rubber vulcanization process. After LC separation using reversed-phase conditions, AgBF(4) in acetonitrile was added, and strong signals were observed for silica-rubber coupling agents and products of the reaction between these and alkenes. The method performs best for substances containing sulfur chains with chain lengths between two and eight sulfur atoms, but sulfur-free compounds containing triethoxysilyl groups were detected as well. For the latter, the postcolumn addition of NaBF(4) proved to be a suitable alternative. Besides the coupling agents, various reaction products, including sulfur-chain bridged alkenes were identified.

  6. Determination of methylparaben from cosmetic products by ultra performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    MANUELA M. MINCEA

    2009-06-01

    Full Text Available A new method for the determination of methylparaben by ultra-performance liquid chromatography (UPLC was developed. Methylparaben is often used as preservative, alone or in combination with other parabens, being added to cosmetic products, pharmaceutical products and foods to avoid microbial contamination. Due to its widespread use and potential risk to human health, assessing human exposure to this compound is of interest. A good determination and quantification of methylparaben was developed with a gradient elution using a mixture of methanol and water (60:40, v/v within 1.455 min. Under optimized conditions, the linear working range extends over two orders of magnitude with relative standard deviations of intra- and inter-day precision below 2.3 %, and a detection limit of 0.02 ng μL-1 for methylparaben. The proposed method was successfully applied to the assay of methylparaben in cosmetic products with minimal sample preparation.

  7. Rifampicin Interference in the Measurement of Urinary Catecholamines by High-Performance Liquid Chromatography.

    Science.gov (United States)

    Kim, Hyun-Ki; Ko, Dae-Hyun; Jeong, Tae-Dong; Lee, Sang-Hee; Lee, Woochang; Lee, Soo-Youn; Chun, Sail; Song, Junghan; Min, Won-Ki

    2015-01-01

    Measurement of urine and plasma catecholamines and their metabolites are essential for the diagnosis of pheochromocytoma. Here, we report our experience of rifampicin interference in the measurement of urinary catecholamines using high-performance liquid chromatography. During a 1-year period, four patients taking rifampicin showed unusual chromatograms with markedly larger norepinephrine spikes, translating to highly elevated norepinephrine concentrations: 290.2, 720.1, 312.0, and 812.7 μg/L. This interference effect did not occur when samples were prepared using the alumina method, with norepinephrine values decreasing to 26.4, 31.5, 21.9, and 17.2 μg/L. Interference was also absent in data using LC-MS/MS. Clinical laboratories should verify whether rifampicin interferes with test results in their assays, and physicians should be informed that urinary norepinephrine levels can be falsely elevated in patients taking rifampicin. © 2015 by the Association of Clinical Scientists, Inc.

  8. [Systematic evaluation of retention behavior of carbohydrates in hydrophilic interaction liquid chromatography].

    Science.gov (United States)

    Fu, Qing; Wang, Jun; Liang, Tu; Xu, Xiaoyong; Jin, Yu

    2013-11-01

    A systematic evaluation of retention behavior of carbohydrates in hydrophilic interaction liquid chromatography (HILIC) was performed. The influences of mobile phase, stationary phase and buffer salt on the retention of carbohydrates were investigated. According to the results, the retention time of carbohydrates decreased as the proportion of acetonitrile in mobile phase decreased. Increased time of carbohydrates was observed as the concentration of buffer salt in mobile phase increased. The retention behavior of carbohydrates was also affected by organic solvent and HILIC stationary phase. Furthermore, an appropriate retention equation was used in HILIC mode. The retention equation lnk = a + blnC(B) + cC(B) could quantitatively describe the retention factors of carbohydrates of plant origin with good accuracy: the relative error of the predicted time to actual time was less than 0.3%. The evaluation results could provide guidance for carbohydrates to optimize the experimental conditions in HILIC method development especially for carbohydrate separation

  9. Rapid and sensitive liquid chromatography-mass spectrometry method for determination of ropinirole in human plasma.

    Science.gov (United States)

    Bhatt, Jignesh; Jangid, Arvind; Shetty, Raghavendra; Shah, Bhavin; Kambli, Sandeep; Subbaiah, Gunta; Singh, Sadhana

    2006-03-18

    A rapid and robust liquid chromatography-mass spectrometry (LC-MS/MS) method was developed for non-ergoline dopamine D(2)-receptor agonist, ropinirole in human plasma using Es-citalopram oxalate as an internal standard. The method involves solid phase extraction from plasma, reversed-phase simple isocratic chromatographic conditions and mass spectrometric detection that enables a detection limit at picogram levels. The proposed method was validated with linear range of 20-1,200 pg/ml. The extraction recoveries for ropinirole and internal standard were 90.45 and 65.42%, respectively. The R.S.D.% of intra-day and inter-day assay was lower than 15%. For its sensitivity and reliability, the proposed method is particularly suitable for pharmacokinetic studies.

  10. Determination of salvinorins and divinatorins in Salvia divinorum leaves by liquid chromatography/multistage mass spectrometry.

    Science.gov (United States)

    Medana, Claudio; Massolino, Cristina; Pazzi, Marco; Baiocchi, Claudio

    2006-01-01

    Salvinorin A is the most potent naturally occurring hallucinogen known and rivals synthetic LSD in potency. Structurally it belongs to the neoclerodane diterpenoids, and it is the only known non-nitrogenous kappa-opioid-selective agonist. Salvia divinorum (Diviner's sage) is a member of the mint family that was used in ancient Mexican traditional practices. Today it is widely cultivated in Europe as a recreational marijuana substitute; it is illegal to buy, sell or possess the plant or the active principle in some countries. Six different salvinorins and three divinatorins have been isolated from Salvia divinorum leaves. The ion fragmentation, separation and quantitation of these diterpenes by liquid chromatography/electrospray ionization multistage mass spectrometry (LC/ESI-MS(n)) are described. The importance of LC in herbal extract determination and the chemical diagnostic power of MS(n) in the analysis of classes of natural organic products are discussed.

  11. Uncovering Biologically Significant Lipid Isomers with Liquid Chromatography, Ion Mobility Spectrometry and Mass Spectrometry

    Science.gov (United States)

    Kyle, Jennifer E.; Zhang, Xing; Weitz, Karl K.; Monroe, Matthew E.; Ibrahim, Yehia M.; Moore, Ronald J.; Cha, Jeeyeon; Sun, Xiaofei; Lovelace, Erica S.; Wagoner, Jessica; Polyak, Stephen J.; Metz, Thomas O.; Dey, Sudhansu K.; Smith, Richard D.; Burnum-Johnson, Kristin E.; Baker, Erin S.

    2016-01-01

    Understanding how biological molecules are generated, metabolized and eliminated in living systems is important for interpreting processes such as immune response and disease pathology. While genomic and proteomic studies have provided vast amounts of information over the last several decades, interest in lipidomics has also grown due to improved analytical technologies revealing altered lipid metabolism in type 2 diabetes, cancer, and lipid storage disease. Mass spectrometry (MS) measurements are currently the dominant approach for characterizing the lipidome by providing detailed information on the spatial and temporal composition of lipids. However, interpreting lipids’ biological roles is challenging due to the existence of numerous structural and stereoisomers (i.e. distinct acyl chain and double-bond positions), which are often unresolvable using present approaches. Here we show that combining liquid chromatography (LC) and structurally-based ion mobility spectrometry (IMS) measurement with MS analyses distinguishes lipid isomers and allows insight into biological and disease processes. PMID:26734689

  12. Identification of microorganisms grown in blood culture flasks using liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Berendsen, Erwin M; Levin, Evgeni; Braakman, René; der Riet-van Oeveren, Debora van; Sedee, Norbert Ja; Paauw, Armand

    2017-10-01

    Bloodstream infections are a common cause of disease and a fast and accurate identification of the causative agent or agents of bloodstream infections would aid the start of adequate treatment. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) shotgun proteomics method was developed for the identification of bacterial species directly from blood cultures that were simulated by inoculating blood culture bottles with single or multiple clinically relevant microorganisms. Using LC-MS/MS, the single species were correctly identified in 100% of the blood cultures, whereas for polymicrobial infections, 78% of both species were correctly identified in blood cultures. The LC-MS/MS method allows for the identification of the causative agent of positive blood cultures.

  13. [Determination of aspirin and free salicylic acid in lysinipirine injection by high performance liquid chromatography].

    Science.gov (United States)

    Dong, Yu; Zhao, Yuan-zheng; Zhang, Yi-na

    2002-05-01

    The contents of aspirin and free salicylic acid in lysinipirine injection were determined by high performance liquid chromatography (HPLC). A Hypersil BDS C18 column was used with the mobile phase of methanol-water-acetic acid (35:65:3, volume ratio) and the detection wavelength of 280 nm. The average recoveries of aspirin and salicylic acid added were 99.27% (RSD = 0.8%) and 99.61%(RSD = 1.3%), respectively. The calibration curves had good linearity in the range of 0.028 g/L -0.141 mg/L and 0.77 mg/L -3.85 mg/L, and the correlation coefficients were 0.9999 and 0.9998 for aspirin and salicylic acid respectively.

  14. Equilibrium theory analysis of liquid chromatography with non-constant velocity.

    Science.gov (United States)

    Ortner, Franziska; Joss, Lisa; Mazzotti, Marco

    2014-12-19

    In liquid chromatography, adsorption and desorption lead to velocity variations within the column if the adsorbing compounds make up a high volumetric ratio of the mobile phase and if there is a substantial difference in the adsorption capacities. An equilibrium theory model for binary systems accounting for these velocity changes is derived and solved analytically for competitive Langmuir isotherms. Characteristic properties of concentration and velocity profiles predicted by the derived model are illustrated by two exemplary systems. Applicability of the model equations for the estimation of isotherm parameters from experimental data is investigated, and accurate results are obtained for systems with one adsorbing and one inert compound, as well as for systems with two adsorbing compounds. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. First screening method for the simultaneous detection of seven allergens by liquid chromatography mass spectrometry.

    Science.gov (United States)

    Heick, J; Fischer, M; Pöpping, B

    2011-02-18

    The development of a multi-method for the detection of seven allergens based on liquid chromatography and triple-quadrupole tandem mass spectrometry in multiple reaction mode is described. It is based on extraction of the allergenic proteins from a food matrix, followed by enzymatic digestion with trypsin. The chosen marker peptides were implemented into one method that is capable of the simultaneous detection of milk, egg, soy, hazelnut, peanut, walnut and almond. This method has been used to detect all seven allergenic commodities from incurred reference bread material, which was baked according to a standard recipe from the baking industry. Detected concentrations ranged from 10 to 1000 μg/g, demonstrating that the mass spectrometric based method is a useful tool for allergen screening. Copyright © 2010 Elsevier B.V. All rights reserved.

  16. Calcium isotope fractionation in liquid chromatography with crown ether resin in methanol medium.

    Science.gov (United States)

    Nemoto, Shigeki; Nomura, Masao; Oi, Takao

    2013-06-01

    Breakthrough mode liquid chromatography was employed to investigate calcium (Ca) isotope fractionation in methanol medium. Highly porous silica beads, the inner pores of which were embedded with a benzo-18-crown-6 ether resin, were used as column packing material. Enrichment of heavier isotopes of Ca was observed in the frontal part of the respective Ca chromatograms. The values of the isotope fractionation coefficient (ϵ) were in the order of 10(-3) at 25 °C. Use of methanol as solvent has little advantage over the aqueous system as far as the values of ϵ are concerned. However, a substantial improvement was observed concerning the adsorption capacity of the crown ether resin for Ca ions. Molecular orbital calculations supported the present isotopic results in a qualitative fashion.

  17. Determination of primary amino acids in wines by high performance liquid magneto-chromatography.

    Science.gov (United States)

    Barrado, E; Rodriguez, J A; Castrillejo, Y

    2009-05-15

    Eight amino acids (ethanolamine, glycine, alanine, beta-aminobutyric acid, leucine, methionine, histidine and asparagine) were identified and quantified in Spanish wines by high performance liquid magneto-chromatography (HPLMC) with UV-V spectrophotometry. For this method, the amino acids are first complexed with mono(1,10-phenanthroline)-Cu(II) to confer them paramagnetic properties, and then separated by application of a low magnetic field intensity (5.5 mT) to the stationary phase contained in the chromatographic column. Principal components analysis of the results obtained grouped together the wine samples according to their denomination of origin: "Ribera del Duero", "Rueda" or "Rioja" (Spain). Through cluster analysis, a series of correlations was also observed among certain amino acids, and between these groupings and the type of wine. These clusters were found to reflect the role played by the amino acids as primary or secondary nutrients for the bacteria involved in alcoholic and malolactic fermentation.

  18. Electrochemically-modulated liquid chromatography (EMLC): Column design, retention processes, and applications

    Energy Technology Data Exchange (ETDEWEB)

    Ting, En -Yi [Iowa State Univ., Ames, IA (United States)

    1997-10-08

    This work describes the continued development of a new separation technique, electrochemically-modulated liquid chromatography (EMLC), from column design, retention mechanisms to pharmaceutical applications. The introduction section provides a literature review of the technique as well as a brief overview of the research in each of the chapters. This section is followed by four chapters which investigate the issues of EMLC column design, the retention mechanism of monosubstituted aromatic compounds, and the EMLC-based applications to two important classes of pharmaceutical compounds (i.e., corticosteroids and benzodiazepines). These four sections have been removed to process separately for inclusion on the database. The dissertation concludes with a general summary, a prospectus, and a list of references cited in the General Introduction. 32 refs.

  19. Optimization of the separation of some psychotropic drugs and their respective metabolites by liquid chromatography.

    Science.gov (United States)

    Cutroneo, P; Beljean, M; Luu, R Phan Tan; Siouffi, A-M

    2006-05-03

    A chemometric procedure is described to optimize the separation of some drugs used in the treatment of psychotic disorders: haloperidol, levomepromazine, risperidone, venlafaxine, carbamazepine and their main metabolites: reduced haloperidol, 9-hydroxy risperidone, desmethyl levomepromazine, desmethyl venlafaxine. The purpose of the procedure is the unambiguous identification and detection in biological fluids. Isocratic reversed-phase liquid chromatography with diode array detection was utilized. An experimental design methodology was carried out in which the experimental response is selectivity. In this way the designs for mixture compounds and for process variables (five variables) was performed which produced 36 experiments to carry out. The desirability function was used to select optimum separation conditions. The procedure provides a chromatogram of well separated solutes.

  20. High performance liquid chromatography method for the determination of cinnamyl alcohol dehydrogenase activity in soybean roots.

    Science.gov (United States)

    dos Santos, W D; Ferrarese, Maria de Lourdes Lucio; Ferrarese-Filho, O

    2006-01-01

    This study proposes a simple, quick and reliable method for determining the cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) activity in soybean (Glycine max L. Merr.) roots using reversed-phase high performance liquid chromatography (RP-HPLC). The method includes a single extraction of the tissue and conduction of the enzymatic reaction at 30 degrees C with cinnamaldehydes (coniferyl or sinapyl), substrates of CAD. Disappearance of the substrates in the reaction mixture is monitored at 340 nm (for coniferaldehyde) or 345 nm (for sinapaldehyde) by isocratic elution with methanol/acetic acid through a GLC-ODS (M) column. This HPLC technique furnishes a rapid and reliable measure of cinnamaldehyde substrates, and may be used as an alternative tool to analyze CAD activity in enzyme preparation without previous purification.

  1. Triacylglycerol profiling of microalgae strains for biofuel feedstock by liquid chromatography-high-resolution mass spectrometry.

    Science.gov (United States)

    MacDougall, Karen M; McNichol, Jesse; McGinn, Patrick J; O'Leary, Stephen J B; Melanson, Jeremy E

    2011-11-01

    Biofuels from photosynthetic microalgae are quickly gaining interest as a viable carbon-neutral energy source. Typically, characterization of algal feedstock involves breaking down triacylglycerols (TAG) and other intact lipids, followed by derivatization of the fatty acids to fatty acid methyl esters prior to analysis by gas chromatography (GC). However, knowledge of the intact lipid profile could offer significant advantages for discovery stage biofuel research such as the selection of an algal strain or the optimization of growth and extraction conditions. Herein, lipid extracts from microalgae were directly analyzed by ultra-high pressure liquid chromatography-mass spectrometry (UHPLC-MS) using a benchtop Orbitrap mass spectrometer. Phospholipids, glycolipids, and TAGs were analyzed in the same chromatographic run, using a combination of accurate mass and diagnostic fragment ions for identification. Using this approach, greater than 100 unique TAGs were identified over the six algal strains studied and TAG profiles were obtained to assess their potential for biofuel applications. Under the growth conditions employed, Botryococcus braunii and Scenedesmus obliquus yielded the most comprehensive TAG profile with a high abundance of TAGs containing oleic acid.

  2. Organic monolith frits encased in polyether ether ketone tubing with improved durability for liquid chromatography.

    Science.gov (United States)

    Park, Sin Young; Cheong, Won Jo

    2015-09-01

    This study introduces a preparation method for polymer-encased monolith frits with improved durability for liquid chromatography columns. The inner surface of the polyether ether ketone tubing is pretreated with sulfuric acid in the presence of catalysts (vanadium oxide and sodium sulfate). The tubing was rinsed with water and acetone, flushed with nitrogen, and treated with glycidyl methacrylate. After washing, the monolith reaction mixture composed of lauryl methacrylate, ethylene glycol dimethacrylate, initiator, and porogenic solvent was filled in the tubing and subjected to in situ polymerization. The tubing was cut into thin slices and used as frits for microcolumns. To check their durability, the frit slices were placed in a vial and a heavy impact was applied on the vial by a vortex mixer for various periods. The frits made in the presence of catalysts were found to be more durable than those made without catalysts. Furthermore, when the monolith-incorporated tubing was used as a chromatography column, the column prepared in the presence of catalysts resulted in a better separation efficiency. The separation performance of the columns installed with the polyether ether ketone encased monolith frits was comparable to that of the columns installed with the commercial stainless-steel screen frits. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Triacylglycerol profiling of microalgae strains for biofuel feedstock by liquid chromatography-high-resolution mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    MacDougall, Karen M.; McNichol, Jesse; McGinn, Patrick J.; O' Leary, Stephen J.B.; Melanson, Jeremy E. [Institute for Marine Biosciences, National Research Council of Canada, Halifax, NS (Canada)

    2011-11-15

    Biofuels from photosynthetic microalgae are quickly gaining interest as a viable carbon-neutral energy source. Typically, characterization of algal feedstock involves breaking down triacylglycerols (TAG) and other intact lipids, followed by derivatization of the fatty acids to fatty acid methyl esters prior to analysis by gas chromatography (GC). However, knowledge of the intact lipid profile could offer significant advantages for discovery stage biofuel research such as the selection of an algal strain or the optimization of growth and extraction conditions. Herein, lipid extracts from microalgae were directly analyzed by ultra-high pressure liquid chromatography-mass spectrometry (UHPLC-MS) using a benchtop Orbitrap mass spectrometer. Phospholipids, glycolipids, and TAGs were analyzed in the same chromatographic run, using a combination of accurate mass and diagnostic fragment ions for identification. Using this approach, greater than 100 unique TAGs were identified over the six algal strains studied and TAG profiles were obtained to assess their potential for biofuel applications. Under the growth conditions employed, Botryococcus braunii and Scenedesmus obliquus yielded the most comprehensive TAG profile with a high abundance of TAGs containing oleic acid. (orig.)

  4. Determination of streptomycin residues in honey by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Granja, Rodrigo H M M; Niño, Alfredo M Montes; Zucchetti, Roberto A M; Niño, Rosario E Montes; Patel, Raj; Salerno, Alessandro G

    2009-04-01

    Streptomycin is an aminoglycoside antibiotic used in apiculture to protect bees against a variety of brood diseases. Brazilian authorities have included it in the National Regulatory Monitoring Program for honey production. A simple and reliable method using liquid chromatography-tandem mass spectrometry has been developed and validated for the determination of streptomycin in honey. The chromatography separation was performed on a Gemini 5 microm C18 (50 mm x 2 mm) column using 5mM heptafluorbutiric acid/acetonitrile (85:15) as the mobile phase at a flow rate of 0.2 mLmin(-1). The detection of the analyte was achieved by positive ionization electrospray in multiple reaction-monitoring modes. Two characteristic transitions were monitored for streptomycin. Some analytical parameters were validated according to the guidelines laid down by European Commission Decision 2002/657/EC: decision limit, detection capability, recovery, precision and ruggedness. The recoveries of streptomycin from honey fortified at 2.5, 10, 15 and 20 microgkg(-1) levels are around 100%. The decision limit and detection capability of streptomycin was 3 microgkg(-1) and 4.7 microgkg(-1) respectively.

  5. Application of hight performance liquid chromatography with spectrofluorimetric detection for determination of human milk retinol

    Directory of Open Access Journals (Sweden)

    Falahat Pisheh HR

    2001-11-01

    Full Text Available World health organization has introduced concentration of vitamin A in breast milk as a proper indicator for characterization of vitamin A deficiency in lactating mothers and their infants (<6 mo. A normal phase high performance liquid chromatography with spectrofluorometric detection as a specific and sensitive detector was used for characterization of all-trans retional from 13-Cis retinal. The method was sensitive (0.009 ppm and accurate (1±0.054 µ mol/l and had a good recovery percentage (99.36%. This method was more better than before methods and was compatible with the other useful methods. Effect of freezing and defreezing on all-trans retinol content of milk sample was investigated. The result was interesting, whatever all-trans retinol content was bigger, it’s destroying was bigger too. Because of that reseon, day today reproducibility wasn’t good. Standard of retinol was 95% all-trans and pure so we should use this method because two retinol isomers wasn’t separated from each other by reversed phase chromatography and UV detection.

  6. Chemistry and liquid chromatography methods for the analyses of primary oxidation products of triacylglycerols.

    Science.gov (United States)

    Zeb, A

    2015-05-01

    Triacylglycerols (TAGs) are one of the major components of the cells in higher biological systems, which can act as an energy reservoir in the living cells. The unsaturated fatty acid moiety is the key site of oxidation and formation of oxidation compounds. The TAG free radical generates several primary oxidation compounds. These include hydroperoxides, hydroxides, epidioxides, hydroperoxy epidioxides, hydroxyl epidioxides, and epoxides. The presence of these oxidized TAGs in the cell increases the chances of several detrimental processes. For this purpose, several liquid chromatography (LC) methods were reported in their analyses. This review is therefore focused on the chemistry, oxidation, extraction, and the LC methods reported in the analyses of oxidized TAGs. The studies on thin-layer chromatography were mostly focused on the total oxidized TAGs separation and employ hexane as major solvent. High-performance LC (HPLC) methods were discussed in details along with their merits and demerits. It was found that most of the HPLC methods employed isocratic elution with methanol and acetonitrile as major solvents with an ultraviolet detector. The coupling of HPLC with mass spectrometry (MS) highly increases the efficiency of analysis as well as enables reliable structural elucidation. The use of MS was found to be helpful in studying the oxidation chemistry of TAGs and needs to be extended to the complex biological systems.

  7. Ionic liquid-regenerated macroporous cellulose monolith: Fabrication, characterization and its protein chromatography.

    Science.gov (United States)

    Du, Kaifeng

    2017-04-21

    Macroporous cellulose monolith as chromatographic support was successfully fabricated from an ionic liquid dissolved cellulose solution by an emulsification method and followed by the cross-linking reaction and DEAE modification. With the physical characterization, the cellulose monolith featured by both the interconnected macropores in range of 0.5-2.5μm and the diffusion pores centered at about 10nm. Given the bimodal pore system, the monolith possessed the specific surface area of 36.4m 2 g -1 and the column permeability of about 7.45×10 -14 m 2 . After the DEAE modification, the anion cellulose monolith was evaluated for its chromatography performances. It demonstrated that the static and dynamic adsorption capacity of BSA reached about 66.7mgmL -1 and 43.9mgmL -1 at 10% breakthrough point, respectively. The results were comparable to other chromatographic adsorbent. In addition, the proteins mixture with different pI was well separated at high flow velocity (611.0cmh -1 ) and high protein recovery (over 97%), proving the macroporous cellulose monolith had excellent separation performance. In this way, the prepared cellulose monolith with bimodal pores system is expected for the potential application in high-speed chromatography. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Simulating Retention in Gas-Liquid Chromatography: Benzene, Toluene, and Xylene Solutes

    Energy Technology Data Exchange (ETDEWEB)

    WICK,COLLIN D.; MARTIN,MARCUS G.; SIEPMANN, J. ILJA; SCHURE,MARK R.

    2000-07-12

    Accurate predictions of retention times, retention indices, and partition constants are a long sought-after goal for theoretical studies in chromatography. Although advances in computational chemistry have improved the understanding of molecular interactions, little attention has been focused on chromatography, let alone calculations of retention properties. Configurational-bias Monte Carlo simulations in the isobaric-isothermal Gibbs ensemble were used to investigate the partitioning of benzene, toluene, and the three xylene isomers between a squalane liquid phase and a helium vapor phase. The united-atom representation of the TraPPE (Transferable Potentials for Phase Equilibria) force field was used for all solutes and squalane. The Gibbs free energies of transfer and Kovats retention indices of the solutes were calculated directly from the partition constants (which were averaged over several independent simulations). While the calculated Kovats indices of benzene and toluene at T = 403 K are significantly higher than their experimental counterparts, much better agreement is found for the xylene isomers at T = 365 K.

  9. Isolation and purification of alkaloids from lotus leaves by ionic-liquid-modified high-speed countercurrent chromatography.

    Science.gov (United States)

    Wu, Nan; Xie, Huihui; Fang, Yingtong; Liu, Yuanyuan; Xi, Xingjun; Chu, Qiao; Dong, Genlai; Lan, Tao; Wei, Yun

    2017-10-31

    An effective high-speed countercurrent chromatography method was successfully established by using ionic liquids as the modifier of the two-phase solvent system. Adding a small amount of ionic liquids significantly shortens the separation time and improves the separation efficiency. The conditions of ionic-liquid-modified high-speed countercurrent chromatography including solvent systems, types and content of added ionic liquids, and ionic liquids post-treatment were investigated. The established method was successfully applied to separate alkaloids from lotus leaves using a two-phase solvent system composed of petroleum ether/ethyl acetate/methanol/water/[C4 mim][BF4 ] (1:5:1:5:0.15, v/v). Four alkaloids pronuciferine (1.7 mg), N-nornuciferine (4.3 mg), nuciferine (3.1 mg), and roemerine (2.1 mg) were obtained with the purities of 90.53, 92.25, 99.86 and 98.63%, respectively from 100 mg crude extract of lotus leaves. The results indicated that ionic-liquid-modified high-speed countercurrent chromatography method was suitable for alkaloid separation from lotus leaves and would be a promising method for the separation of alkaloids from other natural products. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  10. Purification of human albumin by the combination of the method of Cohn with liquid chromatography

    Directory of Open Access Journals (Sweden)

    Tanaka K.

    1998-01-01

    Full Text Available Large volumes of plasma can be fractionated by the method of Cohn at low cost. However, liquid chromatography is superior in terms of the quality of the product obtained. In order to combine the advantages of each method, we developed an integrated method for the production of human albumin and immunoglobulin G (IgG. The cryoprecipitate was first removed from plasma for the production of factor VIII and the supernatant of the cryoprecipitate was fractionated by the method of Cohn. The first precipitate, containing fractions (F-I + II + III, was used for the production of IgG by the chromatographic method (see Tanaka K et al. (1998 Brazilian Journal of Medical and Biological Research, 31: 1375-1381. The supernatant of F-I + II + III was submitted to a second precipitation and F-IV was obtained and discarded. Albumin was obtained from the supernatant of the precipitate F-IV by liquid chromatography, ion-exchange on DEAE-Sepharose FF, filtration through Sephacryl S-200 HR and introduction of heat treatment for fatty acid precipitation. Viral inactivation was performed by pasteurization at 60ºC for 10 h. The albumin product obtained by the proposed procedure was more than 99% pure for the 15 lots of albumin produced, with a mean yield of 25.0 ± 0.5 g/l plasma, containing 99.0 to 99.3% monomer, 0.7 to 1.0% dimers, and no polymers. Prekallikrein activator levels were <=5 IU/ml. This product satisfies the requirements of the 1997 Pharmacopée Européenne.

  11. Simultaneous quantification of various retinoids by high performance liquid chromatography: its relevance to alcohol research.

    Science.gov (United States)

    Yokoyama, H; Matsumoto, M; Shiraishi, H; Ishii, H

    2000-04-01

    We established a high performance liquid chromatography system that allowed simultaneous quantification of various retinoids. We applied the retinoids to a high performance liquid chromatography system with a silica gel absorption column. Samples were separated by the system with a binary multistep gradient with two kinds of solvent that contained n-Hexan, 2-propanol, and glacial acetic acid in different ratios. Each retinoid was detected at a wavelength of 350 nm. This condition allowed separation of 13-cis-retinoic acid, 9-cis-retinoic acid, all-trans-retinoic acid, 13-cis-retinol, all-trans-retinol, all-trans-4-oxo-retinoic acid, and 13-cis-4-oxo-retinoic acid as distinct single peaks. Each retinoid was also analyzed separately and its retention time determined. To ascertain the reliability of this system for retinoid quantification, retinoids at various concentrations were applied to the system. We observed the linearities between the concentration and area under the curve of the peak for each retinoid by linear least-squares regression analysis up to 2.5 ng/ml for all retinoic acids and up to 5 ng/ml for all retinols. There was no significant scattering in tests of within-day reproducibility or day-to-day reproducibility. Using this system, we examined effects of light exposure on isomerization of retinoids. When retinoids were exposed to room light for 2 hr, the amounts of all but 13-cis-retinol changed significantly. In particular, the amounts of all-trans-retinoic acid and 9-cis-retinoic acid were reduced by 40% and 60%, respectively. The HPLC system established in this study should be useful for studying the oxidation pathway of retinol to retinoic acid. A light-shielded condition is required when particular retinoic acids are analyzed.

  12. Measurement of teicoplanin by liquid chromatography-tandem mass spectrometry: development of a novel method.

    Science.gov (United States)

    Fung, Francis H Y; Tang, Jonathan C Y; Hopkins, John P P; Dutton, John J; Bailey, Lisa M; Davison, Andrew S

    2012-09-01

    Teicoplanin is an antibiotic used for the treatment of endocarditis, osteomyelitis, septic arthritis and methicillin-resistant Staphylococcus aureus. Teicoplanin is emerging as a suitable alternative antibiotic to vancomycin, where their trough serum levels are monitored by immunoassay routinely. This is the first report detailing the development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for measuring teicoplanin in patients' serum. An Acquity™ UPLC (ultra-pressure liquid chromatography) tandem mass spectrometer was used to measure teicoplanin concentrations in samples from patients, quality assurance schemes and quality control preparations. Ristocetin was successfully implemented as a suitable internal standard. Ion suppression, linearity, stability, matrix effects, recovery, imprecision, lower limits of quantification and detection, interference and method comparison against immunoassay were all assessed. Teicoplanin and ristocetin had elution times of 1.39 and 1.24 min, respectively. Ion suppression was shown to be negligible, and linear calibration curves (0-200 μg/mL) were consistently reproduced to have r(2) values >0.99. Postextraction stability was achieved up to 20 h, while matrix effects were minimal coupled with sample recovery of >93%. The lower limit of quantification was 1 μg/mL, and 0.2 μg/mL was the lower limit of detection. Interference with other antibiotics was dependent on the combination of drugs present in patients' serum. A method comparison between immunoassay and LC-MS/MS suggested a negative bias for tandem mass spectrometry. This novel method of teicoplanin determination by LC-MS/MS is proven to be a robust protocol that is consistent and reproducible. Clinicians searching for alternatives in therapeutic drug monitoring may have an additional option that is potentially more accurate and specific.

  13. Aniline-modified porous graphitic carbon for hydrophilic interaction and attenuated reverse phase liquid chromatography.

    Science.gov (United States)

    Iverson, Chad D; Lucy, Charles A

    2014-12-19

    Most stationary phases for hydrophilic interaction liquid chromatography (HILIC) and reversed phase liquid chromatography (RPLC) are based on silica. Porous graphitic carbon (PGC) is an attractive alternative to silica-based phases due to its chemical and thermal stability, and unique selectivity. However, native PGC is strongly hydrophobic and in some instances excessively retentive. PGC particles with covalently attached aniline groups (Dimethylaniline-PGC and Aniline-PGC) were synthesized to alter the surface polarity of PGC. First, the diazonium salt of N,N-dimethyl-p-phenylenediamine or 4-nitroaniline was adsorbed onto the PGC surface. The adsorbed salt was reduced with sodium borohydride and (Aniline-PGC only) the nitro group was further reduced with iron powder to the aniline. X-ray photoelectron spectroscopy confirmed the surface functionalities and that these moieties were introduced to the surface at concentrations of 0.9 and 2.1molecules/nm(2), respectively. These modified PGC phases (especially Aniline-PGC) were evaluated as HILIC and reversed phases. The Dimethylaniline-PGC phase displayed only weak HILIC retention of phenolic solutes. In contrast, the Aniline-PGC phase displayed up to nearly a 7-fold increase in HILIC retention vs. an aniline-silica phase and selectivity that differed from 10 other HILIC phases. Introduction of aniline groups to the PGC surface reduced the RPLC retentivity of PGC up to more than 5-fold and improved the separation efficiency up to 6-fold. The chromatographic performance of Aniline-PGC is demonstrated by separations of nucleotides, nucleosides, carboxylic acids, basic pharmaceuticals, and other compounds. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Quantification of liquid products from the electroreduction of CO2 and CO using static headspace-gas chromatography and nuclear magnetic resonance spectroscopy

    DEFF Research Database (Denmark)

    Bertheussen, Erlend; Abghoui, Younes; Jovanov, Zarko P.

    2017-01-01

    Static headspace-gas chromatography (HS-GC) useful for ex-situ liquid product analysis. Could complement high-performance liquid chromatography and NMR spectroscopy. Particularly high sensitivity towards compounds with high vapor pressure. Detection limits below 0.5μM were shown for acetaldehyde ...

  15. Multi-detection of corticosteroids in sports doping and veterinary control using high-resolution liquid chromatography/time-of-flight mass spectrometry

    NARCIS (Netherlands)

    Touber, M.E.; Engelen, M.C.; Georgakopoulus, C.; Rhijn, van J.A.; Nielen, M.W.F.

    2007-01-01

    A liquid chromatography/time-of-flight mass spectrometry (LC/TOFMS) method was developed using the latest high-resolution LC column technology, the ultra performance liquid chromatography (UPLC (TM)), and electrospray ionization (ESI) in the positive ion mode. Gradient UPLC separation conditions

  16. Near-infrared laser-induced fluorescence detection in column liquid chromatography. A comparison of various lasers and detection systems. 1. Continuous wave lasers.

    NARCIS (Netherlands)

    Mank, A.J.G.; Velthorst, N.H.; Brinkman, U.A.T.; Gooijer, C.

    1995-01-01

    Several lasers, i.e. HeNe lasers, diode lasers and an argon-ion-dye laser combination, are compared as excitation sources for near-infrared laser-induced fluorescence detection in column liquid chromatography. Using a model gradient liquid chromatography system, detection limits for disulphonated

  17. Quantitative analysis of cytokinins in plants by high performance liquid chromatography: electronspray ionization ion trap mass spectrometry.

    Science.gov (United States)

    Chen, Weiqi; Gai, Ying; Liu, Shichang; Wang, Renxiao; Jiang, Xiangning

    2010-10-01

    The present paper introduces a highly sensitive and selective method for simultaneous quantification of 12 cytokinins (free form and their conjugates). The method includes a protocol of extraction with methanol/water/formic acid (15/4/1, v/v/v) to the micro-scale samples, pre-purification with solid phase extraction (SPE) cartridges of the extracts, separation with a high performance liquid chromatography (HPLC) and detection by an electrospray ionization ion trap mass spectrometry (ESI-Ion trap-MS) system in a consecutive ion monitoring (CRM) mode at the three stage fragmentation of mass spectrometry (MS(3) ). The lowest detection level of the cytokinins of the method reaches 0.1-2.0 pg with a very wide range of linear regression from 1-512 pg, at the coefficient factors of 0.98-0.99. The feasibility of this method has been proven in the application of the method to the analysis of the trace-amount contents of cytokinins in the micro-scale samples of various types of plant materials, such as aerial parts of rice and poplar leaves etc. 12 endogenous cytokinins had been identified and quantified in the plant tissues, with an acceptable relatively higher recovery rate from 40% to 70%. © 2010 Institute of Botany, Chinese Academy of Sciences.

  18. Determination of erythromycins in fermentation broth using liquid phase extraction with back extraction combined with high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Fahimeh Kamarei

    2014-07-01

    Full Text Available Liquid phase extraction with back extraction (LPE-BE combined with high performance liquid chromatography-diode array detection (HPLC-DAD was applied for the extraction and determination of erythromycin A, B and C in fermentation broths. According to this procedure, the fermentation broth with the adjustment pH at a fixed value of 10 was first mixed with organic solvent (Vbroth/Vorg = 1.0. After shaking, the mixture was separated into two phases by microfuging at 13,000 rpm for 15 min. Then back extraction was performed into the acidic aqueous phase with pH 5.0 (Vorg/Vaq = 1.0. After centrifugation at 3000, the two phases were separated and 50 μL of the acidic aqueous phase was injected into the HPLC. The effects of different variables such as the nature of extraction solvent and the pH of samples and buffer were investigated. At the most appropriate conditions, dynamic linear ranges of 0.5–8, 0.1–0.9 and 0.1–0.9 mg mL−1 and limits of detection of 0.03, 0.003 and 0.002 mg mL−1 were obtained for erythromycin A, B and C, respectively. Relative standard deviations (RSDs of the proposed method were less than 9.5%. The mean recoveries were 99.5%. The proposed method is simple and sensitive with highly clean-up effect and it can be used for monitoring the progress of erythromycin fermentation.

  19. Intravascular Residence Time Determination for the Cyanide Antidote Dimethyl Trisulfide in Rat by Using Liquid-Liquid Extraction Coupled with High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Deepthika De Silva

    2016-01-01

    Full Text Available These studies represent the first report on the intravascular residence time determinations for the cyanide antidote dimethyl trisulfide (DMTS in a rat model by using high performance liquid chromatography coupled with ultraviolet absorption spectroscopy (HPLC-UV. The newly developed sample preparation included liquid-liquid extraction by cyclohexanone. The calibration curves showed a linear response for DMTS concentrations between 0.010 and 0.30 mg/mL with R2 = 0.9994. The limit of detection for DMTS via this extraction method was 0.010 mg/mL, and the limit of quantitation was 0.034 mg/mL. Thus this calibration curve provided a tool for determining DMTS in the range between 0.04 and 0.30 mg/mL. Rats were given 20 mg/kg DMTS dose (in 15% Polysorbate 80 intravenously, and blood samples were taken 15, 60, 90, 120, and 240 min after DMTS injections. The data points were plotted as DMTS concentration in RBCs versus time, and the intravascular residence time was determined graphically. The results indicated a half-life of 36 min in a rat model, suggesting that the circulation time is long enough to provide a reasonable time interval for cyanide antagonism.

  20. A rapid ultrasound-assisted dispersive liquid-liquid microextraction followed by ultra-performance liquid chromatography for the simultaneous determination of seven benzodiazepines in human plasma samples.

    Science.gov (United States)

    Fernández, Purificación; González, Cristina; Pena, M Teresa; Carro, Antonia M; Lorenzo, Rosa A

    2013-03-12

    A simple and efficient ultrasound-assisted dispersive liquid-liquid microextraction (UA-DLLME) method has been developed for the determination of seven benzodiazepines (alprazolam, bromazepam, clonazepam, diazepam, lorazepam, lormetazepam and tetrazepam) in human plasma samples. Chloroform and methanol were used as extractant and disperser solvents, respectively. The influence of several variables (e.g., type and volume of dispersant and extraction solvents, pH, ultrasonic time and ionic strength) was carefully evaluated and optimized, using an asymmetric screening design 3(2)4(2)//16. Analysis of extracts was performed by ultra-performance liquid chromatography coupled with photodiode array detection (UPLC-PDA). Under the optimum conditions, two reversed-phases, Shield RP18 and C18 columns were successfully tested, obtaining good linearity in a range of 0.01-5μgmL(-1), with correlation coefficients r>0.996. Quantification limits ranged between 4.3-13.2ngmL(-1) and 4.0-14.8ngmL(-1), were obtained for C18 and Shield RP18 columns, respectively. The optimized method exhibited a good precision level, with relative standard deviation values lower than 8%. The recoveries studied at two spiked levels, ranged from 71 to 102% for all considered compounds. The proposed method was successfully applied to the analysis of seven benzodiazepines in real human plasma samples. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. [Determination of capsaicinoids and eugenol in waste-edible-oil by liquid-liquid extraction and liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Zhang, Zhong; Ren, Fei; Zhang, Pan

    2012-11-01

    A method was developed for the determination of capsaicinoids (capsaicin, dihydrocapsaicin and synthetic capsaicin) and eugenol in waste-edible-oil extracted by liquid-liquid extraction and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The capsaicinoids and eugenol in waste-edible-oil were extracted by methanol, and then separated by a SUPEL COSIL ABZ + Plus dC18 column (150 mm x4.6 mm, 5 microm). The analysis was performed by MS/MS with electrospray ionization in positive and negative ion modes with multiple reaction monitoring (MRM). The limits of detection for capsaicin, dihydrocapsaicin, synthetic capsaicin and eugenol were 0.02, 0.03, 0.03 and 0.6 microg/L, respectively. The good linear relationships were obtained in certain concentration ranges of capsaicinoids and eugenol. The relative standard deviations (RSDs, n=5) of same-worker and different-worker were less than 5%. The method is exclusive, sensitive and accurate, and can be used in waste-edible-oil determination.

  2. Determination of benzophenone-3 and its main metabolites in human serum by dispersive liquid-liquid microextraction followed by liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Tarazona, Isuha; Chisvert, Alberto; Salvador, Amparo

    2013-11-15

    A new analytical method for the determination of benzophenone-3 (2-hydroxy-4-methoxybenzophenone), and its main metabolites (2,4-dihydroxybenzophenone and 2,2'-dihydroxy-4-methoxybenzophenone) in human serum is presented. The method is based on dispersive liquid-liquid microextraction (DLLME) as preconcentration and clean-up technique, followed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Acidic hydrolysis and protein precipitation with HCl 6 M (1:1) (100 °C, 1 h) were carried out before extraction. The variables involved in the DLLME process were studied. Under the optimized conditions, 70 µL of acetone (disperser solvent) and 30 µL of chloroform (extraction solvent) were mixed and rapidly injected into 800 µL of hydrolyzed serum sample. Sample pH or ionic strength adjustment were not necessary. The method was validated by analyzing spiked human serum samples. No satisfactory recoveries were obtained when aqueous standards or standards prepared in synthetic serum were used, but excellent recoveries were achieved by using matrix-matched calibration standards. Moreover, limits of detection in the low µg L(-1) level and good repeatability were obtained. In order to show the applicability of the proposed method in the study of percutaneous absorption processes, it was applied to the analysis of serum samples from two volunteers after topical application of a sunscreen cosmetic product containing 2-hydroxy-4-methoxybenzophenone. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Dispersive Liquid-Liquid Microextraction Combined with Ultrahigh Performance Liquid Chromatography/Tandem Mass Spectrometry for Determination of Organophosphate Esters in Aqueous Samples

    Directory of Open Access Journals (Sweden)

    Haiying Luo

    2014-01-01

    Full Text Available A new technique was established to identify eight organophosphate esters (OPEs in this work. It utilised dispersive liquid-liquid microextraction in combination with ultrahigh performance liquid chromatography/tandem mass spectrometry. The type and volume of extraction solvents, dispersion agent, and amount of NaCl were optimized. The target analytes were detected in the range of 1.0–200 µg/L with correlation coefficients ranging from 0.9982 to 0.9998, and the detection limits of the analytes were ranged from 0.02 to 0.07 µg/L (S/N=3. The feasibility of this method was demonstrated by identifying OPEs in aqueous samples that exhibited spiked recoveries, which ranged between 48.7% and 58.3% for triethyl phosphate (TEP as well as between 85.9% and 113% for the other OPEs. The precision was ranged from 3.2% to 9.3% (n=6, and the interprecision was ranged from 2.6% to 12.3% (n=5. Only 2 of the 12 selected samples were tested to be positive for OPEs, and the total concentrations of OPEs in them were 1.1 and 1.6 µg/L, respectively. This method was confirmed to be simple, fast, and accurate for identifying OPEs in aqueous samples.

  4. Low-density solvent-based dispersive liquid-liquid microextraction followed by high performance liquid chromatography for determination of warfarin in human plasma.

    Science.gov (United States)

    Ghambari, Hoda; Hadjmohammadi, Mohammadreza

    2012-06-15

    Extraction and determination of warfarin, a widely used anticoagulant drug, in human plasma were performed using a new generation of dispersive liquid-liquid microextraction (DLLME) and high performance liquid chromatography (HPLC). The extraction procedure is based on extraction solvents lighter than water and performing of extraction in a specially designed extraction cell. Some important parameters, including kind and volume of extraction and disperser solvents, pH of the sample solution, salt concentration in the sample solution and extraction time were investigated and optimized. Under the optimized conditions (150 μL 1-octanol as extraction solvent, 150 μL methanol as disperser solvent, pH(sample)=2.3, extraction time of 2 min, without salt addition), limit of detection (LOD) of 5 ng mL⁻¹ and extraction recovery of 91.0% were obtained. The calibration curve was linear within the range of 15-3000 ng mL⁻¹ with the square of correlation coefficient (R²) of 0.998. Repeatability and reproducibility of method based on five replicate extraction and determination were 2.8% and 6.5%, respectively. The proposed method was applied successfully for the determination of warfarin in plasma sample from a patient under treatment with this drug, and was demonstrated to be sensitive, efficient, and convenient. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Vortex-assisted low density solvent based demulsified dispersive liquid-liquid microextraction and high-performance liquid chromatography for the determination of organophosphorus pesticides in water samples.

    Science.gov (United States)

    Seebunrueng, Ketsarin; Santaladchaiyakit, Yanawath; Srijaranai, Supalax

    2014-05-01

    A simple, rapid, effective and eco-friendly preconcentration method, vortex-assisted low density solvent based solvent demulsified dispersive liquid-liquid microextraction (VLDS-SD-DLLME), followed by high performance liquid chromatography-diode array detector (HPLC-DAD) analysis, has been developed for the first time for the determination of four organophosphorus pesticides (OPPs) (e.g., azinphos-methyl, parathion-methyl, fenitrothion and diazinon) in environmental water samples. In this preconcentration procedure, an emulsion was obtained after the mixture of extraction solvent (1-dodecanol) and dispersive solvent (acetonitrile, ACN) was injected rapidly into 10 mL of the sample solution. The vortex agitator aided the dispersion of the extraction solvent into the sample solution. After the formation of an emulsion, the demulsifier (ACN) was added, resulting in the rapid separation of the mixture into two phases without centrifugation. Under optimal conditions, the proposed method provided high extraction efficiency (90-99%), good linearity range (0.5-500 ng mL(-1)), low limits of detection (0.25-1 ng mL(-1)) and good repeatability and recoveries were obtained. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Determination of estrogenic mycotoxins in environmental water samples by low-toxicity dispersive liquid-liquid microextraction and liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Emídio, Elissandro Soares; da Silva, Claudia Pereira; de Marchi, Mary Rosa Rodrigues

    2015-04-24

    A novel, simple, rapid and eco-friendly method based on dispersive liquid-liquid microextraction using a bromosolvent was developed to determine six estrogenic mycotoxins (zearalenone, zearalanone, α-zearalanol, β-zearalanol, α-zearalenol and β-zearalenol) in water samples by liquid chromatography-electrospray ionization tandem mass spectrometry in the negative mode (LC-ESI-MS/MS). The optimal conditions for this method include the use of 100 μL bromocyclohexane as an extraction solvent (using a non-dispersion solvent), 10 mL of aqueous sample (adjusted to pH 4), a vortex extraction time of 2 min, centrifugation for 10 min at 3500 rpm and no ionic strength adjustment. The calibration function was linear and was verified by applying the Mandel fitting test with a 95% confidence level. No matrix effect was observed. According to the relative standard deviations (RSDs), the precision was better than 13% for the repeatability and intermediate precision. The average recoveries of the spiked compounds ranged from 81 to 118%. The method limits of detection (LOD) and quantification (LOQ) considering a 125-fold pre-concentration step were 4-20 and 8-40 ng L(-1), respectively. Next, the method was applied to the analysis of the environmental aqueous samples, demonstrating the presence of β-zearalanol and zearalanone in the river water samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Automated dispersive liquid-liquid microextraction coupled to high performance liquid chromatography - cold vapour atomic fluorescence spectroscopy for the determination of mercury species in natural water samples.

    Science.gov (United States)

    Liu, Yao-Min; Zhang, Feng-Ping; Jiao, Bao-Yu; Rao, Jin-Yu; Leng, Geng

    2017-04-14

    An automated, home-constructed, and low cost dispersive liquid-liquid microextraction (DLLME) device that directly coupled to a high performance liquid chromatography (HPLC) - cold vapour atomic fluorescence spectroscopy (CVAFS) system was designed and developed for the determination of trace concentrations of methylmercury (MeHg(+)), ethylmercury (EtHg(+)) and inorganic mercury (Hg(2+)) in natural waters. With a simple, miniaturized and efficient automated DLLME system, nanogram amounts of these mercury species were extracted from natural water samples and injected into a hyphenated HPLC-CVAFS for quantification. The complete analytical procedure, including chelation, extraction, phase separation, collection and injection of the extracts, as well as HPLC-CVAFS quantification, was automated. Key parameters, such as the type and volume of the chelation, extraction and dispersive solvent, aspiration speed, sample pH, salt effect and matrix effect, were thoroughly investigated. Under the optimum conditions, linear range was 10-1200ngL(-1) for EtHg(+) and 5-450ngL(-1) for MeHg(+) and Hg(2+). Limits of detection were 3.0ngL(-1) for EtHg(+) and 1.5ngL(-1) for MeHg(+) and Hg(2+). Reproducibility and recoveries were assessed by spiking three natural water samples with different Hg concentrations, giving recoveries from 88.4-96.1%, and relative standard deviations <5.1%. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Vortex-assisted liquid-liquid micro-extraction and high-performance liquid chromatography for a higher sensitivity methyl methacrylate determination in biological matrices.

    Science.gov (United States)

    Sousa, Tiago F A; Aniceto, Marta C; Amorim, Célia G; Souto-Lopes, Mariana; Pérez-Mongiovi, Daniel; Montenegro, Maria C B S M; Araújo, Alberto N

    2014-05-01

    A vortex-assisted liquid-liquid micro-extraction coupled with high-performance liquid chromatography, with UV-vis, is proposed to pre-concentrate methyl methacrylate and to improve separation in biological matrices. The use of 1-octanol as extracting phase, its volume, the need for a dispersant agent, the agitation conditions and the cooling time before phase separation were evaluated. In optimum conditions, enrichment factors of 20 (±0.5) and enrichment recovery of 99% were obtained. The straightforward association of this extraction process with the HPLC method, previously regulated by the International Organization for Standardization, afforded a detection limit of 122 ng/mL and a quantification limit of 370 ng/mL. The within-batch precision, relative standard deviation, was 3% for a sample with 1.49 µg/mL and 4% for a sample with 13.4 µg/mL. The results showed a between batch-precision of 21% for experiments performed on five different days, for a sample with a concentration of 1.10 µg/mL in methyl methacrylate. Copyright © 2013 John Wiley & Sons, Ltd.

  9. Dispersive liquid-liquid microextraction for the determination of three cytokinin compounds in fruits and vegetables by liquid chromatography with time-of-flight mass spectrometry.

    Science.gov (United States)

    Campillo, N; Viñas, P; Férez-Melgarejo, G; Hernández-Córdoba, M

    2013-11-15

    The paper presents a novel approach for the determination of three cytokinin compounds, thidiazuron (TDZ), 1,3-diphenylurea (1,3-DPU) and forchlorfenuron (CPPU), in fruit and vegetables samples using liquid chromatography with electrospray ionization and time-of-flight mass spectrometry (LC-ESI-TOFMS). Analytes were extracted from the sample matrix with ethanol, and the extract, after dilution with water, was submitted to dispersive liquid-liquid microextraction (DLLME). Once acetonitrile and 1,2-dichloroethane had been selected as extraction and disperser solvents, respectively, the influence of the following experimental parameters was studied using a Plackett-Burman design: volume of extraction and disperser solvents, sample mass and time and speed of centrifugation. The best analytical conditions were 250 µL 1,2-dichloroethane, 1.5 mL acetonitrile, 5 g sample mass, and centrifugation at 3000 rpm for 3 min. The optimized method provided DLs in the range 0.02-0.05 ng g(-1), depending on the compound. Satisfactory recovery values between 89 and 106% were obtained for spiked samples (kiwifruit, watermelon, grape and tomato) in the 0.2-1.0 ng g(-1) concentration range, depending on the compound. None of the target analytes was detected in any of the samples analyzed. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Determination of Fusarium toxins in functional vegetable milks applying salting-out-assisted liquid-liquid extraction combined with ultra-high-performance liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Hamed, Ahmed M; Arroyo-Manzanares, Natalia; García-Campaña, Ana M; Gámiz-Gracia, Laura

    2017-11-01

    Vegetable milks are considered as functional foods due to their physiological benefits. Although the consumption of these products has significantly increased, they have received little attention in legislation with regard to contaminants. However, they may contain mycotoxins resulting from the use of contaminated raw materials. In this work, ultra-high-performance liquid chromatography tandem mass spectrometry has been proposed for the determination of the most relevant Fusarium toxins (fumonisin B1 and B2, HT-2 and T-2 toxins, zearalenone, deoxynivalenol and fusarenon-X) in different functional beverages based on cereals, legumes and seeds. Sample treatment consisted of a simple salting-out-assisted liquid-liquid extraction with no further clean-up. The method provided limits of quantification between 3.2 and 57.7 µg L-1, recoveries above 80% and precision with RSD lower than 12%. The method was also applied for studying the occurrence of these mycotoxins in market samples of vegetable functional beverages and deoxynivalenol was found in three oat-based commercial drinks.

  11. Sensitivity enhancement by chromatographic peak concentration with ultra-high performance liquid chromatography-nuclear magnetic resonance spectroscopy for minor impurity analysis.

    Science.gov (United States)

    Tokunaga, Takashi; Akagi, Ken-Ichi; Okamoto, Masahiko

    2017-07-28

    High performance liquid chromatography can be coupled with nuclear magnetic resonance (NMR) spectroscopy to give a powerful analytical method known as liquid chromatography-nuclear magnetic resonance (LC-NMR) spectroscopy, which can be used to determine the chemical structures of the components of complex mixtures. However, intrinsic limitations in the sensitivity of NMR spectroscopy have restricted the scope of this procedure, and resolving these limitations remains a critical problem for analysis. In this study, we coupled ultra-high performance liquid chromatography (UHPLC) with NMR to give a simple and versatile analytical method with higher sensitivity than conventional LC-NMR. UHPLC separation enabled the concentration of individual peaks to give a volume similar to that of the NMR flow cell, thereby maximizing the sensitivity to the theoretical upper limit. The UHPLC concentration of compound peaks present at typical impurity levels (5.0-13.1 nmol) in a mixture led to at most three-fold increase in the signal-to-noise ratio compared with LC-NMR. Furthermore, we demonstrated the use of UHPLC-NMR for obtaining structural information of a minor impurity in a reaction mixture in actual laboratory-scale development of a synthetic process. Using UHPLC-NMR, the experimental run times for chromatography and NMR were greatly reduced compared with LC-NMR. UHPLC-NMR successfully overcomes the difficulties associated with analyses of minor components in a complex mixture by LC-NMR, which are problematic even when an ultra-high field magnet and cryogenic probe are used. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Integrated Graduate and Continuing Education in Protein Chromatography for Bioprocess Development and Scale-Up

    Science.gov (United States)

    Carta, Jungbauer

    2011-01-01

    We describe an intensive course that integrates graduate and continuing education focused on the development and scale-up of chromatography processes used for the recovery and purification of proteins with special emphasis on biotherapeutics. The course includes lectures, laboratories, teamwork, and a design exercise and offers a complete view of…

  13. Quantitative profiling of tryptophan metabolites in serum, urine, and cell culture supernatants by liquid chromatography-tandem mass spectrometry

    NARCIS (Netherlands)

    Zhu, Wentao; Stevens, Axel P.; Dettmer, Katja; Gottfried, Eva; Hoves, Sabine; Kreutz, Marina; Holler, Ernst; Canelas, Andre B.; Kema, Ido; Oefner, Peter J.

    2011-01-01

    A sensitive, selective, and comprehensive method for the quantitative determination of tryptophan and 18 of its key metabolites in serum, urine, and cell culture supernatants was developed. The analytes were separated on a C18 silica column by reversed-phase liquid chromatography and detected by

  14. Simultaneous Determination of Caffeine and Vitamin B6 in Energy Drinks by High-Performance Liquid Chromatography (HPLC)

    Science.gov (United States)

    Leacock, Rachel E.; Stankus, John J.; Davis, Julian M.

    2011-01-01

    A high-performance liquid chromatography experiment to determine the concentration of caffeine and vitamin B6 in sports energy drinks has been developed. This laboratory activity, which is appropriate for an upper-level instrumental analysis course, illustrates the standard addition method and simultaneous determination of two species. (Contains 1…

  15. Identification and quantification of cannabinoids in Cannabis sativa L. plants by high performance liquid chromatography-mass spectrometry

    NARCIS (Netherlands)

    Aizpurua-Olaizola, Oier; Omar, Jone; Navarro, Patricia; Olivares, Maitane; Etxebarria, Nestor; Usobiaga, Aresatz

    2014-01-01

    High performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) has been successfully applied to cannabis plant extracts in order to identify cannabinoid compounds after their quantitative isolation by means of supercritical fluid extraction (SFE). MS conditions were optimized by means

  16. Determination of methyldibromoglutaronitrile in cosmetic products by high-performance liquid chromatography with electrochemical detection. Method validation

    DEFF Research Database (Denmark)

    Rastogi, Suresh Chandra; Zachariae, Claus; Johansen, Jeanne D

    2004-01-01

    An increased frequency of contact allergy to methyldibromoglutaronitrile (MDBGN), a commonly used preservative in cosmetics and other consumer products, has been reported in recent years. A high-performance liquid chromatography (HPLC) method for the determination of MDBGN in cosmetic products ha...

  17. Chemically modified polymeric resins for separation of cations, organic acids, and small polar moleculea by high performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Morris, John B. [Iowa State Univ., Ames, IA (United States)

    1993-07-01

    This thesis is divided into 4 parts: a review, ion chromatography of metal cations on carboxylic resins, separation of hydrophilic organic acids and small polar compounds on macroporous resin columns, and use of eluent modifiers for liquid chromatographic separation of carboxylic acids using conductivity detection.

  18. Determination of the brominated flame retardant, hexabroomcyclodocane in sediments and biota by liquid chromatography-electrospray ionization mass spectrometry

    NARCIS (Netherlands)

    Morris, S.; Bersuder, P.; Allchin, C.R.; Zegers, B.; Boon, J.P.; Leonards, P.E.G.; Boer, de J.

    2006-01-01

    A method involving reversed-phase, liquid chromatography coupled to electrospray ionisation mass spectrometry (LC-ESI-MS) was developed for separation, detection and quantitation of the alpha-, beta- and gamma-diastereoisomers of hexabromocyclododecane (HBCD). To address the lack of environmental

  19. Three-mode factor analysis of data on retention in normal-phase high-performance liquid chromatography

    NARCIS (Netherlands)

    Ligny, C.L. de; Spanjer, M.C.; Houwelingen, J.C. van; Weesie, H.M.

    1984-01-01

    It is shown that the Snyder equation is not quite satisfactory for fitting retention data in normal-phase high-performance liquid chromatography (HPLC) on chemically bonded phases. This equation is a special case of the mathematical—statistical three-mode factor analysis model. This model, in its

  20. Comprehensive analysis of pharmaceutical products using simultaneous mixed-mode (ion-exchange/reversed-phase) and hydrophilic interaction liquid chromatography.

    Science.gov (United States)

    Kazarian, Artaches A; Nesterenko, Pavel N; Soisungnoen, Phimpha; Burakham, Rodjana; Srijaranai, Supalax; Paull, Brett

    2014-08-01

    Liquid chromatographic assays were developed using a mixed-mode column coupled in sequence with a hydrophilic interaction liquid chromatography column to allow the simultaneous comprehensive analysis of inorganic/organic anions and cations, active pharmaceutical ingredients, and excipients (carbohydrates). The approach utilized dual sample injection and valve-mediated column switching and was based upon a single high-performance liquid chromatography gradient pump. The separation consisted of three distinct sequential separation mechanisms, namely, (i) ion-exchange, (ii) mixed-mode interactions under an applied dual gradient (reversed-phase/ion-exchange), and (iii) hydrophilic interaction chromatography. Upon first injection, the Scherzo SS C18 column (Imtakt) provided resolution of inorganic anions and cations under isocratic conditions, followed by a dual organic/salt gradient to elute active pharmaceutical ingredients and their respective organic counterions and potential degradants. At the top of the mixed-mode gradient (high acetonitrile content), the mobile phase flow was switched to a preconditioned hydrophilic interaction liquid chromatography column, and the standard/sample was reinjected for the separation of hydrophilic carbohydrates, some of which are commonly known excipients in drug formulations. The approach afforded reproducible separation and resolution of up to 23 chemically diverse solutes in a single run. The method was applied to investigate the composition of commercial cough syrups (Robitussin®), allowing resolution and determination of inorganic ions, active pharmaceutical ingredients, excipients, and numerous well-resolved unknown peaks. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Accurate quantitation of pentaerythritol tetranitrate and its degradation products using liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry

    NARCIS (Netherlands)

    Brust, H.; Asten, A. van; Koeberg, M.; Dalmolen, J.; Heijden, A.E.D.M. van der; Schoenmakers, P.

    2014-01-01

    After an explosion of pentaerythritol tetranitrate (PETN), its degradation products pentaerythritol trinitrate (PETriN), dinitrate (PEDiN) and mononitrate (PEMN) were detected using liquid chromatography-atmospheric-pressure chemical-ionization-mass spectrometry (LC-APCI-MS). Discrimination between

  2. Comparative urine analysis by liquid chromatography-mass spectrometry and multivariate statistics : Method development, evaluation, and application to proteinuria

    NARCIS (Netherlands)

    Kemperman, Ramses F. J.; Horvatovich, Peter L.; Hoekman, Berend; Reijmers, Theo H.; Muskiet, Frits A. J.; Bischoff, Rainer

    2007-01-01

    We describe a platform for the comparative profiling of urine using reversed-phase liquid chromatography-mass spectrometry (LC-MS) and multivariate statistical data analysis. Urinary compounds were separated by gradient elution and subsequently detected by electrospray Ion-Trap MS. The lower limit

  3. Structural Elucidation of DNA-Protein Crosslinks Using Reductive Desulfurization and Liquid Chromatography-Tandem Mass Spectrometry

    Science.gov (United States)

    Wickramaratne, Susith

    2014-01-01

    Structural characterization of DNA-protein crosslinks involving cysteine using reductive desulfurization in combination with liquid chromatography-tandem mass spectrometry is highlighted. The novel approach was used to identify hydrolytically stable DNA-protein lesions involving alkylguanine DNA alkyltransferase (AGT). PMID:24436288

  4. Development of on-line liquid chromatography-biochemical detection for soluble epoxide hydrolase inhibitors in mixtures.

    NARCIS (Netherlands)

    Falck, D.; Schebb, N.H.; Prinatiningtyas, S; Zhang, Jiawen; Heus, F.A.H.; Morisseau, C; Kool, J.; Hammock, B.D.; Niessen, W.M.A.

    2013-01-01

    In this study, an end-point-based fluorescence assay for soluble epoxide hydrolase (sEH) was transformed into an on-line continuous-flow format. The on-line biochemical detection system (BCD) was coupled on-line to liquid chromatography (LC) to allow mixture analysis. The on-line BCD was based on a

  5. Constructing a LabVIEW-Controlled High-Performance Liquid Chromatography (HPLC) System: An Undergraduate Instrumental Methods Exercise

    Science.gov (United States)

    Smith, Eugene T.; Hill, Marc

    2011-01-01

    In this laboratory exercise, students develop a LabVIEW-controlled high-performance liquid chromatography system utilizing a data acquisition device, two pumps, a detector, and fraction collector. The programming experience involves a variety of methods for interface communication, including serial control, analog-to-digital conversion, and…

  6. Sensitive absorption detection for micro-column liquid chromatography by ultraviolet forward-scattering degenerate four-wave mixing.

    NARCIS (Netherlands)

    de Beer, T.; Beso, P.; Velthorst, N.H.; Brinkman, U.A.T.; Gooijer, C.; Hoornweg, G.Ph.

    1999-01-01

    The performance of forward-scattering degenerate four-wave mixing (F-D4WM) in the mid-ultraviolet (UV) region (351nm) as a detection technique for micro-column liquid chromatography (μLC) is studied, using nitro-substituted polycyclic aromatic hydrocarbons (NO

  7. Liquid chromatography-tandem mass spectrometry assay for the quantification of free and total sialic acid in human cerebrospinal fluid.

    NARCIS (Netherlands)

    Ham, M. van der; Koning, T.J. de; Lefeber, D.J.; Fleer, A.; Prinsen, B.H.; Sain-van der Velden, M.G. de

    2010-01-01

    BACKGROUND: Analysis of sialic acid (SA) metabolites in cerebrospinal fluid (CSF) is important for clinical diagnosis. In the present study, a high-performance liquid chromatography-tandem mass spectrometry (HPLC/MS/MS) method for free sialic acid (FSA) and total sialic acid (TSA) in human CSF was

  8. Liquid chromatography-tandem mass spectrometry assay for the quantification of free and total sialic acid in human cerebrospinal fluid

    NARCIS (Netherlands)

    van der Ham, Maria; de Koning, Tom J; Lefeber, Dirk; Fleer, André; Prinsen, Berthil H C M T; de Sain-van der Velden, Monique G M

    2010-01-01

    BACKGROUND: Analysis of sialic acid (SA) metabolites in cerebrospinal fluid (CSF) is important for clinical diagnosis. In the present study, a high-performance liquid chromatography-tandem mass spectrometry (HPLC/MS/MS) method for free sialic acid (FSA) and total sialic acid (TSA) in human CSF was

  9. Simultaneous determination of estrogens and progestogens in honey using high performance liquid chromatography-tandem mass spectrometry

    Science.gov (United States)

    This work describes the development and validation of a method for the simultaneous determination of 13 estrogens and progestogens in honey by high performance liquid chromatography-tandem mass spectrometry. The target compounds were preconcentrated by solid phase extraction. Pretreatment variables ...

  10. Cavity ring-down spectroscopy for detection in liquid chromatography: Extension to tunable sources and ultraviolet wavelengths

    NARCIS (Netherlands)

    van der Sneppen, L.; Wiskerke, A.E.; Ariese, F.; Gooijer, C.; Ubachs, W.M.G.

    2006-01-01

    In earlier studies, it was demonstrated that the sensitivity of absorbance detection in liquid chromatography (LC) can be improved significantly by using cavity ring-down spectroscopy (CRDS). Thus far, CRDS experiments have been performed using visible laser light at fixed standard wavelengths, such

  11. Determination of the HCV protease inhibitor telaprevir in plasma and dried blood spot by liquid chromatography-tandem mass spectrometry

    NARCIS (Netherlands)

    Verweij-Van Wissen, Corrien P W G M; De Graaff-Teulen, Marga J A; De Kanter, Clara T M M; Aarnoutse, Rob E.; Burger, David M.

    2015-01-01

    Background: Telaprevir is a protease inhibitor used in the treatment of hepatitis C virus infection. Analytical methods for telaprevir should separate the compound from its R-diastereomer VRT-127394, which is 30-fold less active. The objective of this work was to develop liquid chromatography-tandem

  12. Determination of the HCV Protease Inhibitor Telaprevir in Plasma and Dried Blood Spot by Liquid Chromatography-Tandem Mass Spectrometry

    NARCIS (Netherlands)

    Verweij-van Wissen, C.P.W.G.M.; Graaff-Teulen, M.J.A. de; Kanter, C.T.M.M. de; Aarnoutse, R.E.; Burger, D.M.

    2015-01-01

    BACKGROUND: Telaprevir is a protease inhibitor used in the treatment of hepatitis C virus infection. Analytical methods for telaprevir should separate the compound from its R-diastereomer VRT-127394, which is 30-fold less active. The objective of this work was to develop liquid chromatography-tandem

  13. Rapid analysis of organic microcontaminants in environmental water samples by trace enrichment and liquid chromatography on a single short column.

    NARCIS (Netherlands)

    Hogenboom, A.C.; Malmqvist, U.K.; Nolkrantz, K.; Vreuls, J.J.; Brinkman, U.A.T.

    1997-01-01

    On-column trace enrichment and liquid chromatography using a single short (20 mm length) high-pressure packed column was optimized for the rapid simultaneous identification and quantification of a wide range of organic microcontaminants in environmental water samples. The quality of different C,,

  14. Determination of olanzapine in whole blood using simple protein precipitation and liquid chromatography-tandem mass spectrometry

    DEFF Research Database (Denmark)

    Nielsen, Marie Katrine Klose; Johansen, Sys Stybe

    2009-01-01

    A simple, sensitive, and reproducible liquid chromatography-tandem mass spectrometry method has been developed and validated for the quantification of the antipsychotic drug olanzapine in whole blood using dibenzepine as internal standard (IS). After acidic methanol-induced protein precipitation...

  15. Exploring Liquid Sequential Injection Chromatography to Teach Fundamentals of Separation Methods: A Very Fast Analytical Chemistry Experiment

    Science.gov (United States)

    Penteado, Jose C.; Masini, Jorge Cesar

    2011-01-01

    Influence of the solvent strength determined by the addition of a mobile-phase organic modifier and pH on chromatographic separation of sorbic acid and vanillin has been investigated by the relatively new technique, liquid sequential injection chromatography (SIC). This technique uses reversed-phase monolithic stationary phase to execute fast…

  16. Liquid chromatography-electrospray ionisation-mass spectrometry based method for the determination of estradiol benzoate in hair of cattle

    NARCIS (Netherlands)

    Hooyerink, H.; Lommen, A.; Mulder, P.P.J.; Rhijn, van J.A.; Nielen, M.W.F.

    2005-01-01

    A liquid chromatography (LC)-based method with mass spectrometric (MS/MS) detection was developed for the determination of estradiol benzoate residues in hair of cattle. First hair samples were pulverized with a ball mill followed by treatment with the reducing agent Tris(2-carboxyethyl)phosphine

  17. Solid phase extraction for removal of matrix effects in lipophilic marine toxin analysis by liquid chromatography-tandem mass spectrometry

    NARCIS (Netherlands)

    Gerssen, A.; McElhinney, A. M.; Mulder, P.P.J.; Bire, L.; Hess, P.; de Boer, J.

    2009-01-01

    The potential of solid phase extraction (SPE) clean-up has been assessed to reduce matrix effects (signal suppression or enhancement) in the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of lipophilic marine toxins. A large array of ion-exchange, silica-based, and mixed-function

  18. Solid phase extraction for removal of matrix effects in lipophilic marine toxin analysis by liquid chromatography-tandem mass spectrometry

    NARCIS (Netherlands)

    Gerssen, A.; McElhinney, M.; Mulder, P.P.J.; Bire, R.; Hess, P.; Boer, de J.

    2009-01-01

    The potential of solid phase extraction (SPE) clean-up has been assessed to reduce matrix effects (signal suppression or enhancement) in the liquid chromatography-tandem mass spectrometry (LC¿MS/MS) analysis of lipophilic marine toxins. A large array of ion-exchange, silica-based, and mixed-function

  19. Liquid chromatography coupled with tandem mass spectrometry for the quantitative analysis of anticancer drugs in biological matrices

    NARCIS (Netherlands)

    Stokvis, Ellen

    2004-01-01

    In this thesis, the development and validation of liquid chromatography tandem mass spectrometric (LC-MS/MS) methods for the quantitative bioanalysis of anticancer drugs are described. The monitoring of these drugs in biological fluids and tissues is important during both pre-clinical and clinical

  20. Determination of perfluorocompounds in popcorn packaging by pressurised liquid extraction and ultra-performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Martínez-Moral, María Pilar; Tena, María Teresa

    2012-11-15

    The development and characterisation of a method based on reverse-phase ultra-performance liquid chromatography (UPLC) coupled to a quadrupole-time of flight mass spectrometer (Q-TOF-MS) with negative electrospray ionisation (ESI) to determine perfluorinated compounds (PFCs) in packaging is presented in this paper. Analytes were quantitatively recovered from packaging with methanol in only one PLE cycle of 6 min at 100 °C. The UPLC allowed the successful separation of the studied PFCs in less than 4 min. The whole method presented good precision, with RSDs below 8%, LODs from 0.6 to 16 ng g(-1); and excellent recovery values, around 100% in all cases, were achieved. The PLE-UPLC-MS method was applied to the analysis of popcorn packaging for microwave cooking. Besides the most commonly studied PFCs: PFOA and PFOS, the presence of other perfluorocarboxylic acids (PFCAs) in popcorn packaging is evidenced in this work. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Detection of 10 sweeteners in various foods by liquid chromatography/tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Chui-Shiang Chang

    2014-09-01

    Full Text Available The analytical method for sweeteners in various food matrixes is very important for food quality control and regulation enforcement. A simple and rapid method for the simultaneous determination of 10 sweeteners [acesulfame potassium (ACS-K, aspartame (ASP, cyclamate (CYC, dulcin (DUL, glycyrrhizic acid (GA, neotame (NEO, neohesperidin dihydrochalcone (NHDC, saccharin (SAC, sucralose (SCL, and stevioside (STV] in various foods by liquid chromatography/tandem mass chromatography (LC–MS/MS was developed. The chromatographic separation was performed on a Phenomenex Luna Phenyl-Hexyl (5 μm, 4.6 mm × 150 mm column with gradient elution of 10 mM ammonium acetate in water and 10 mM ammonium acetate in methanol. The recoveries of the 10 sweeteners were between 75% and 120%, and the coefficients of variation were less than 20%. The limits of quantification were 0.5 μg/kg for NHDC and SCL. For the other sweeteners, the limits of quantification were 0.1 μg/kg. Compared to the traditional high-performance liquid chromatography method, the LC–MS/MS method could provide better sensitivity, higher throughput, enhanced specificity, and more sweeteners analyzed in a single run. The samples included 27 beverages (16 alcoholic and 11 nonalcoholic beverages and 15 pickled foods (1 pickled pepper, 3 candies, and 11 candied fruits. Two remanufactured wines were found to contain 7.2, 8.5 μg/g SAC and 126.5, 123 μg/g CYC, respectively. ACS-K, ASP, SCL, and NEO were detected in five beverages and drinks. The pickled peppers and candied fruits were found to contain SAC, GA, CYC, ASP, STV, NEO, and ACS-K. The wine with sweeteners detected was remanufactured wine, not naturally fermented wine. Therefore, the ingredient label for the sweeteners of remanufactured wine should be regulated by the proper authority for inspection of sweeteners.

  2. Supported liquid membrane-liquid chromatography-mass spectrometry analysis of cyanobacterial toxins in fresh water systems

    Science.gov (United States)

    Mbukwa, Elbert A.; Msagati, Titus A. M.; Mamba, Bhekie B.

    Harmful algal blooms (HABs) are increasingly becoming of great concern to water resources worldwide due to indiscriminate waste disposal habits resulting in water pollution and eutrophication. When cyanobacterial cells lyse (burst) they release toxins called microcystins (MCs) that are well known for their hepatotoxicity (causing liver damage) and have been found in eutrophic lakes, rivers, wastewater ponds and other water reservoirs. Prolonged exposure to low concentrated MCs are equally of health importance as they are known to be bioaccumulative and even at such low concentration do exhibit toxic effects to aquatic animals, wildlife and human liver cells. The application of common treatment processes for drinking water sourced from HABs infested reservoirs have the potential to cause algal cell lyses releasing low to higher amounts of MCs in finished water. Trace microcystins in water/tissue can be analyzed and quantified using Liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) following solid-phase extraction (SPE) sample clean-up procedures. However, extracting MCs from algal samples which are rich in chlorophyll pigments and other organic matrices the SPE method suffers a number of drawbacks, including cartridge clogging, long procedural steps and use of larger volumes of extraction solvents. We applied a supported liquid membrane (SLM) based technique as an alternative sample clean-up method for LC-ESI-MS analysis of MCs from both water and algal cells. Four (4) MC variants (MC-RR, -YR, -LR and -WR) from lyophilized cells of Microcystis aeruginosa and water collected from a wastewater pond were identified) and quantified using LC-ESI-MS following a SLM extraction and liquid partitioning step, however, MC-WR was not detected from water extracts. Within 45 min of SLM extraction all studied MCs were extracted and pre-concentrated in approximately 15 μL of an acceptor phase at an optimal pH 2.02 of the donor phase (sample). The highest

  3. Determination of drug lipophilicity by phosphatidylcholine-modified microemulsion high-performance liquid chromatography.

    Science.gov (United States)

    Xuan, Xueyi; Xu, Liyuan; Li, Liangxing; Gao, Chongkai; Li, Ning

    2015-07-25

    A new biomembrane-mimetic liquid chromatographic method using a C8 stationary phase and phosphatidylcholine-modified (PC-modified) microemulsion mobile phase was used to estimate unionized and ionized drugs lipophilicity expressed as an n-octanol/water partition coefficient (logP and logD). The introduction of PC into sodium dodecyl sulfate (SDS) microemulsion yielded a good correlation between logk and logD (R(2)=0.8). The optimal composition of the PC-modified microemulsion liquid chromatography (PC-modified MELC) mobile phase was 0.2% PC-3.0% SDS-6.0% n-butanol-0.8% ethyl acetate-90.0% water (pH 7.0) for neutral and ionized molecules. The interactions between the analytes and system described by this chromatographic method is more similar to biological membrane than the n-octanol/water partition system. The result in this paper suggests that PC-modified MELC can serve as a possible alternative to the shake-flask method for high-throughput unionized and ionized drugs lipophilicity determination and simulation of biological processes. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Characterization of hexacationic imidazolium ionic liquids as effective and highly stable gas chromatography stationary phases.

    Science.gov (United States)

    González-Álvarez, Jaime; Blanco-Gomis, Domingo; Arias-Abrodo, Pilar; Díaz-Llorente, Daniel; Ríos-Lombardía, Nicolás; Busto, Eduardo; Gotor-Fernández, Vicente; Gutiérrez-Álvarez, María D

    2012-01-01

    Polycationic ionic liquids (ILs) are an attractive class of ILs with great potential applicability as gas chromatography stationary phases. A family of hexacationic imidazolium ILs derived from the cycloalkanol family was chemically first prepared in a straightforward manner and then applied for analytical separation purposes. Four tuneable engineering vectors, namely cation ring size structure, anion nature, spatial disposition of cycloalkanol substituents and O-substitution, were considered as experimental parameters for the design of the desired ionic liquids. A total number of five new phases based on a common benzene core respectively exhibited column efficiencies around to 2500 plates/m, broad operating temperature ranges and also, even more importantly, good thermal stabilities (bleeding temperature between 260 and 365°C), finding variations in the selectivity and analytes elution orders depending on the IL structures. Their solvation characteristics were evaluated using the Abraham solvation parameter model, establishing clear correlations between their cation structure and retention capability with respect to certain analytes. The study of relationships between the ILs structure and solvation parameters gives us an idea of the IL stationary phase to be used for specific separations. Copyright © 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Review of online coupling of sample preparation techniques with liquid chromatography.

    Science.gov (United States)

    Pan, Jialiang; Zhang, Chengjiang; Zhang, Zhuomin; Li, Gongke

    2014-03-07

    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.

  6. Reversed-phase high-performance liquid chromatography of tenuazonic acid and related tetramic acids.

    Science.gov (United States)

    Shephard, G S; Thiel, P G; Sydenham, E W; Vleggaar, R; Marasas, W F

    1991-05-03

    A reversed-phase high-performance liquid chromatographic system for the determination of the fungal toxin, tenuazonic acid, (5S,8S)-3-acetyl-5-sec.-butyltetramic acid, is described. The system utilizes a column packed with deactivated end-capped C18 silica with a high carbon load to overcome the problem of poor chromatographic performance of this beta-diketone on reversed-phase liquid chromatography which previously necessitated the use of anion-exchange, ligand-exchange or ion-pairing methods. The reversed-phase system allows the separation of tenuazonic acid from its (5R,8S)-diastereomer, allo-tenuazonic acid and was applied to the detection of tenuazonic acid in cultures of Alternaria alternata and Phoma sorghina. By means of diode-array ultraviolet detection, (5S)-3-acetyl-5-isopropyltetramic acid was observed in extracts of culture material. This metabolite was purified using the analytical reversed-phase system and was identified by 1H and 13C nuclear magnetic resonance spectroscopy.

  7. [Determination of commonly abused dyes in food by liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Yi, Xionghai; Deng, Xiaojun; Yang, Huiqin; Guo, Dehua; Zhu, Jian

    2011-11-01

    A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of commonly abused dyes in candy, sirup, liquid milk and fruit juice. The test sample was dissolved and diluted with water, then cleaned up by polyamide solid phase extraction. The LC separation was performed on an Agilent XDB-C18 column with 20 mmol/L ammonium acetate solution and acetonitrile as the mobile phase in a gradient elution mode. The dyes were determined by MS/MS in negative electrospray ionization mode, and quantified by matrix-matched external standard method. The calibration curves showed good linearity in the range of 0.5 - 50 mg/L with the correlation coefficients (r) greater than 0.99. The limits of quantitation (S/N > 10) were 0.5 mg/kg, and the limits of detection (S/N > 3) were 0.1 mg/kg. The recoveries of dyes in food samples at the three spiked levels of 0.5, 5 and 50 mg/kg were in the range of 62.6% - 115.3% with the relative standard deviations (RSDs) of 2.6% - 26.3%. The method can meet the requirements for the determination of the dyes in food samples for import and export inspection.

  8. High efficiency polyethylene glycol diacrylate monoliths for reversed-phase capillary liquid chromatography of small molecules.

    Science.gov (United States)

    Aggarwal, Pankaj; Lawson, John S; Tolley, H Dennis; Lee, Milton L

    2014-10-17

    Highly cross-linked monolithic networks (i.e., polyethylene glycol diacrylate, PEGDA) synthesized from monomers containing varying ethylene oxide chain lengths were fabricated inside fused silica capillary columns for use in liquid chromatography (LC) of small molecules. Tergitol was used as a surfactant porogen in combination with other typical organic liquid porogens. Column performance was correlated with quantitative descriptors of the physical/chemical properties of the monomers and porogens using a statistical model. Solubility and viscosity values of the components were identified as important predictors of monolith morphology and efficiency. The chromatographic retention mechanism was determined to be principally reversed-phase (RP) with additional hydrogen bonding between the polar groups of the analytes and the ethylene oxide groups embedded in the monolith structure. The fabricated monolithic columns were evaluated under RPLC conditions using phenols, hydroxy benzoic acids, and alkyl parabens as test compounds. Isocratic elution of hydroxy benzoic acids at a linear velocity of 0.04 cm/s using a PEGDA-700 monolith gave chromatographic peaks with little tailing (i.e., tailing factorphenol standards (3 different columns). Copyright © 2014 Elsevier B.V. All rights reserved.

  9. [Determination of phthalic anhydride in ambient air by high performance liquid chromatography].

    Science.gov (United States)

    Zaĭtseva, N V; Ulanova, T S; Karnazhitskaia, T D; Antip'eva, M V; Kislitsina, A V

    2011-01-01

    The paper considers the measurement of phthalic anhydride in the ambient air samples by high performance liquid chromatography. It describes conditions for air sampling and analysis of phthalic anhydride levels in the presence of concomitant components of its production (phthalic and maleic acids, maleic anhydride, etc.) on a liquid chromatograph with an UV detector. The procedure was tested, by estimating the quality of ambient air at the border of a sanitary protection zone of phthalic anhydride production and when analyzing the air in the industrial area. Field studies detected the concentrations of phthalic anhydrate in the air of an enterprise area, which were equal to 0.017-0.115 mg/dm3. Phthalic anhydride was detectable at a concentration of 0.001-0.0021 mg/dm3 at the border of the existing sanitary protection zone in single cases. The procedure has been recommended to measure the mass concentrations of phthalic anhydride aerosol and vapors in ambient air at the reference concentration.

  10. Design and implementation of an automated liquid-phase microextraction-chip system coupled on-line with high performance liquid chromatography

    DEFF Research Database (Denmark)

    Li, Bin; Petersen, Nickolaj J.; Payán, María D Ramos

    2014-01-01

    An automated liquid-phase microextraction (LPME) device in a chip format has been developed and coupled directly to high performance liquid chromatography (HPLC). A 10-port 2-position switching valve was used to hyphenate the LPME-chip with the HPLC autosampler, and to collect the extracted....... The composition of the supported liquid membrane (SLM) and carrier was optimized in order to achieve reasonable extraction performance of all the five alkaloids. With 1-octanol as SLM solvent and with 25mM sodium octanoate as anionic carrier, extraction recoveries for the different opium alkaloids ranged between....... The repeatability was within 5.0-10.8% (RSD). The membrane liquid in the LPME-chip was regenerated automatically between every third injection. With this procedure the liquid membrane in the LPME-chip was stable in 3-7 days depending on the complexity of sample solutions with continuous operation. With this LPME...

  11. Development of a high performance liquid chromatography method and a liquid chromatography-tandem mass spectrometry method with pressurized liquid extraction for simultaneous quantification and confirmation of cyromazine, melamine and its metabolites in foods of animal origin.

    Science.gov (United States)

    Yu, Huan; Tao, Yanfei; Chen, Dongmei; Wang, Yulian; Liu, Zhaoying; Pan, Yuanhu; Huang, Lingli; Peng, Dapeng; Dai, Menghong; Liu, Zhenli; Yuan, Zonghui

    2010-12-03

    Simple and sensitive methods have been developed for simultaneous detection of cyromazine, melamine and their metabolites (ammeline, ammelide and cyanuric acid) in samples of animal origins. These include a high performance liquid chromatography (HPLC) method and a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method and are useful in regular monitoring and in toxicity studies of these molecules. Representative samples used in this study include muscles and livers of swine, bovine, sheep and chicken, kidneys of swine, bovine and sheep, and milk powder. A new sample preparation procedure with pressurized liquid extraction (PLE) at 1400psi and 70°C was investigated. Quantification of these five compounds by HPLC was achieved using an APS-2 column with UV detection at 230 nm. Limit of detection (LOD) was at 10 μgkg(-1), and limit of quantification (LOQ) was at 40 μgkg(-1). Recoveries of the five analytes in spiked samples ranged from 72.2% to 115.4% with RSD less than 12%. Confirmatory analysis of the analytes was performed using LC-MS/MS in selected reaction monitoring (SRM) mode. The LOD and LOQ were 5 μgkg(-1) and 15 μgkg(-1), respectively. This is the first simultaneous analysis of cyromazine, melamine, ammeline, ammelide and cyanuric acid residues in complex tissue samples using PLE and HPLC. It is expected that these methods will find many practical applications in evaluating the safety of cyromazine, melamine and their metabolites. Copyright © 2010 Elsevier B.V. All rights reserved.

  12. Direct total and free testosterone measurement by liquid chromatography tandem mass spectrometry across two different platforms.

    Science.gov (United States)

    Rhea, Jeanne M; French, Deborah; Molinaro, Ross J

    2013-05-01

    To develop and validate liquid chromatography tandem mass spectrometry (LC-MS/MS) methods for the direct measurement of total and free testosterone in patient samples on two different analytical systems. An API 4000 and 5000 triple quadropoles were used and compared; the former is reported to be 3-5 times less sensitive, as was used to set the quantitation limits. Free testosterone was separated from the protein-bound fraction by equilibrium dialysis followed by derivatization. Either free or total testosterone, and a deuterated internal standard (d3-testosterone) were extracted by liquid-liquid extraction. The validation results were compared to two different clinical laboratories. The use of d2-testosterone was found to be unacceptable for our method. The total testosterone LC-MS/MS methods on both systems were linear over a wide concentration range of 1.5-2000ng/dL. Free testosterone was measured directly using equilibrium dialysis coupled LC-MS/MS and linear over the concentration range of 2.5-2500pg/mL. Good correlation (total testosterone, R(2)=0.96; free testosterone, R(2)=0.98) was observed between our LC-MS/MS systems and comparator laboratory. However, differences in absolute values for both free and total testosterone measurements were observed while a comparison to a second published LC-MS/MS method showed excellent correlation. Free and total testosterone measurements correlated well with clinical observations. To our knowledge, this is the first published validation of free and total testosterone methods across two analytical systems of different analytical sensitivities. A less sensitive system does not sacrifice analytical or clinical sensitivity to directly measure free and total testosterone in patient samples. Copyright © 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  13. [Measurement of 11 benzophenone ultraviolet-filters in cosmetics by high performance liquid chromatography].

    Science.gov (United States)

    Qu, Baocheng; Bian, Haitao; Mao, Xiqin; Li, Jin

    2015-12-01

    A sample preparation and analytical method with liquid-liquid extraction (LLE) and high performance liquid chromatography (HPLC) was developed to detect 11 benzophenone ultraviolet-filters in cosmetics. The target compounds were extracted by the mixed solutions of tetrahydrofuran (TH)/methanol/water or dichloromethane/water at proper ratios. The extracts were centrifuged and filtered to remove matrix compounds, and then analyzed by HPLC. The separation of analytes was carried out on a Diamonsil-C18 column (150 mm x 4.6 mm, 5 μm) with 0.1% (v/v) formic acid aqueous solution (containing 10 mmol/L ammonium acetate) as mobile phase A and methanol containing 0.1% (v/v) formic acid as mobile phase B. The spiked recoveries of the method (n = 7) were 93.4%-103.8% with the relative standard deviations of 0.1%-4.2%. The limits of detection (LODs) were in the range of 4.0-30 μg/g and the limits of quantitation (LOQs) ranged from 15 to 100 μg/g. The method was applied to the determination of 42 cosmetic samples randomly purchased from the supermarket in Dalian. Five benzophenone series were always detected, in which the content of benzophenone-3 in sunscreen cream and the content of benzophenone-2 in perfume were very high and reached 2 785 μg/g and 2 106 μg/g, respectively. The results showed that the developed method is efficient, reliable and sensitive, which can be applied to the determination of benzophenones in cosmetics.

  14. Dual reductive/oxidative electrochemistry/liquid chromatography/mass spectrometry: Towards peptide and protein modification, separation and identification.

    Science.gov (United States)

    Büter, Lars; Frensemeier, Lisa M; Vogel, Martin; Karst, Uwe

    2017-01-06

    A new hyphenated technique based on on-line dual (oxidative and reductive) electrochemistry coupled to liquid chromatography and high resolution electrospray mass spectrometry is presented. Two liquid streams are combined, with one containing a disulfide, which is reduced to the respective thiol in an electrochemical cell based on a titanium working electrode. The other stream contains phenol, which is electrochemically activated to benzoquinone on a boron-doped diamond working electrode. Upon combination of the two streams, a Michael addition takes places, leading to the covalent coupling of thiol to quinone. In continuous flow, the reaction mixture is transferred into an injection valve and the products are separated by reversed phase liquid chromatography and detected by electrospray-high resolution mass spectrometry. Proof of concept is demonstrated for low molecular mass disulfides and peptides, but further optimization will be required in future work to achieve efficient protein labelling. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Simulation of elution profiles in liquid chromatography - II: Investigation of injection volume overload under gradient elution conditions applied to second dimension separations in two-dimensional liquid chromatography.

    Science.gov (United States)

    Stoll, Dwight R; Sajulga, Ray W; Voigt, Bryan N; Larson, Eli J; Jeong, Lena N; Rutan, Sarah C

    2017-11-10

    An important research direction in the continued development of two-dimensional liquid chromatography (2D-LC) is to improve the detection sensitivity of the method. This is especially important in applications where injection of large volumes of effluent from the first dimension ( 1 D) column into the second dimension ( 2 D) column leads to severe 2 D peak broadening and peak shape distortion. For example, this is common when coupling two reversed-phase columns and the organic solvent content of the 1 D mobile phase overwhelms the 2 D column with each injection of 1 D effluent, leading to low resolution in the second dimension. In a previous study we validated a simulation approach based on the Craig distribution model and adapted from the work of Czok and Guiochon [1] that enabled accurate simulation of simple isocratic and gradient separations with very small injection volumes, and isocratic separations with mismatched injection and mobile phase solvents [2]. In the present study we have extended this simulation approach to simulate separations relevant to 2D-LC. Specifically, we have focused on simulating 2 D separations where gradient elution conditions are used, there is mismatch between the sample solvent and the starting point in the gradient elution program, injection volumes approach or even exceed the dead volume of the 2 D column, and the extent of sample loop filling is varied. To validate this simulation we have compared results from simulations and experiments for 101 different conditions, including variation in injection volume (0.4-80μL), loop filling level (25-100%), and degree of mismatch between sample organic solvent and the starting point in the gradient elution program (-20 to +20% ACN). We find that that the simulation is accurate enough (median errors in retention time and peak width of -1.0 and -4.9%, without corrections for extra-column dispersion) to be useful in guiding optimization of 2D-LC separations. However, this requires that real

  16. Application and recovery of ionic liquids in the preparative separation of four flavonoids from Rhodiola rosea by on-line three-dimensional liquid chromatography.

    Science.gov (United States)

    Ma, Shufeng; Hu, Liming; Ma, Chaoyang; Lv, Wenping; Wang, Hongxin

    2014-09-01

    A novel on-line three-dimensional liquid chromatography method was developed to separate four main flavonoids from Rhodiola rosea. Ethyl acetate/0.5 mol/L ionic liquid 1-butyl-3-methylimidazolium chloride aqueous solution was selected as the solvent system. In the first-dimension separation, the target flavonoids were entrapped and subsequently desorbed into the second-dimension high-speed countercurrent chromatographic column for separation. In the third-dimension chromatography, the residual ionic liquid in the four separated flavonoids was removed and the used ionic liquid was recovered. As a result, 35.1 mg of compound 1, 20.4 mg of compound 2, 8.5 mg of compound 3, and 10.6 mg of compound 4 were obtained from 1.53 g R. rosea extract. They were identified as rhodiosin, rhodionin, herbacetin, and kaempferol, respectively. The recovery of ionic liquid reached 99.1% of the initial amount. The results showed that this method is a powerful technology for the separation of R. rosea flavonoids and that the ionic-liquid-based solvent system has advantages over traditional solvent systems in renewable and environmentally friendly properties. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. High-performance liquid chromatography separation and purification of cacao (Theobroma cacao L.) procyanidins according to degree of polymerization using a diol stationary phase.

    Science.gov (United States)

    Kelm, Mark A; Johnson, J Chris; Robbins, Rebecca J; Hammerstone, John F; Schmitz, Harold H

    2006-03-08

    A new chromatographic approach for separating cacao procyanidins according to their degree of polymerization has been developed. It utilizes diol stationary phase columns operating in normal phase mode with a binary gradient of acidified acetonitrile and methanol-water. Performance of the diol stationary phase was evaluated on an analytical scale utilizing classical chromatographic conditions for the normal phase separation of procyanidins according to their degree of polymerization. The new separation approach was developed on an analytical scale but further extended to the preparative scale. These newly developed analytical and preparative high-performance liquid chromatography procedures were successfully applied to the separation, as well as isolation, of cacao procyanidins from unfermented cacao seeds. The degree of polymerization associated with each molecular weight fraction was determined by mass spectrometry.

  18. The analysis of carbohydrates in milk powder by a new "heart-cutting" two-dimensional liquid chromatography method.

    Science.gov (United States)

    Ma, Jing; Hou, Xiaofang; Zhang, Bing; Wang, Yunan; He, Langchong

    2014-03-01

    In this study, a new"heart-cutting" two-dimensional liquid chromatography method for the simultaneous determination of carbohydrate contents in milk powder was presented. In this two dimensional liquid chromatography system, a Venusil XBP-C4 analysis column was used in the first dimension ((1)D) as a pre-separation column, a ZORBAX carbohydrates analysis column was used in the second dimension ((2)D) as a final-analysis column. The whole process was completed in less than 35min without a particular sample preparation procedure. The capability of the new two dimensional HPLC method was demonstrated in the determination of carbohydrates in various brands of milk powder samples. A conventional one dimensional chromatography method was also proposed. The two proposed methods were both validated in terms of linearity, limits of detection, accuracy and precision. The comparison between the results obtained with the two methods showed that the new and completely automated two dimensional liquid chromatography method is more suitable for milk powder sample because of its online cleanup effect involved. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

  19. Determination of aflatoxins in medicinal plants by high-performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Siddique, Nadeem A; Mujeeb, Mohd; Ahmad, Sayeed; Panda, Bibhu P; Makhmoor, Mohd

    2013-01-01

    The intention of the proposed work is to study the presence of the aflatoxins B1, B2, G1 and G2 in medicinal plants, namely Mucuna pruriens, Delphinium denudatum and Portulaca oleraceae. The aflatoxins were extracted, purified by immunoaffinity column chromatography and analysed by high-performance liquid chromatography-tandem quadrupole mass spectrometry with electrospray ionisation (HPLC-MS/MS). Fungal count was carried out in PDA media. A good linear relationship was found for AFB1, AFB2, AFG1 and AFG2 at 1-10 ppb (r>0.9995). The analyte accuracy under three different spiking levels was 86.7-108.1 %, with low per cent relative standard deviations in each case. The aflatoxins can be separated within 5 to7 min using an Agilent XDB C18-column. We found that AFB1 and AFB2 were in trace amounts below the detection limit in M. pruriens whilst they were not detected in D. denudatum. P. oleraceae was found to be contaminated with AFB1 and AFB2. AFG1 and AFG2 were not detected in M. pruriens, P. oleraceae and were below the detection limit in D. denudatum. This was consistent with very low numbers of fungal colonies observed after 6 hr of incubation. The analytical method developed is simple, precise, accurate, economical and can be effectively used to determine the aflatoxins in medicinal plants and therefore to control the quality of products. The aflatoxin levels in the plant extracts examined were related to the minimal fungal load in the medicinal plants examined.

  20. Detailed analysis of petroleum hydrocarbon attenuation in biopiles by high-performance liquid chromatography followed by comprehensive two-dimensional gas chromatography.

    Science.gov (United States)

    Mao, Debin; Lookman, Richard; Van De Weghe, Hendrik; Van Look, Dirk; Vanermen, Guido; De Brucker, Nicole; Diels, Ludo

    2009-02-27

    Enhanced bioremediation of petroleum hydrocarbons in two biopiles was quantified by high-performance liquid chromatography (HPLC) followed by comprehensive two-dimensional gas chromatography (GCXGC). The attenuation of 34 defined hydrocarbon classes was calculated by HPLC-GCXGC analysis of representative biopile samples at start-up and after 18 weeks of biopile operation. In general, a-cyclic alkanes were most efficiently removed from the biopiles, followed by monoaromatic hydrocarbons. Cycloalkanes and polycyclic aromatic hydrocarbons (PAHs) were more resistant to degradation. A-cyclic biomarkers farnesane, trimethyl-C13, norpristane, pristane and phytane dropped to only about 10% of their initial concentrations. On the other hand, C29-C31 hopane concentrations remained almost unaltered after 18 weeks of biopile operation, confirming their resistance to biodegradation. They are thus reliable indicators to estimate attenuation potential of petroleum hydrocarbons in biopile processed soils.

  1. Screening of Potential Xanthine Oxidase Inhibitors in Gnaphalium hypoleucum DC. by Immobilized Metal Affinity Chromatography and Ultrafiltration-Ultra Performance Liquid Chromatography-Mass Spectrometry.

    Science.gov (United States)

    Zhang, Hong-Jian; Hu, Yi-Juan; Xu, Pan; Liang, Wei-Qing; Zhou, Jie; Liu, Pei-Gang; Cheng, Lin; Pu, Jin-Bao

    2016-09-17

    In this study, a new method based on immobilized metal affinity chromatography (IMAC) combined with ultrafiltration-ultra performance liquid chromatography-mass spectrometry (UF-UPLC-MS) was developed for discovering ligands for xanthine oxidase (XO) in Gnaphalium hypoleucum DC., a folk medicine used in China for the treatment of gout. By IMAC, the high flavonoid content of G. hypoleucum could be determined rapidly and efficiently. UF-UPLC-MS was used to select the bound xanthine oxidase ligands in the mixture and identify them. Finally, two flavonoids, luteolin-4'-O-glucoside and luteolin, were successfully screened and identified as the candidate XO inhibitors of G. hypoleucum. They were evaluated in vitro for XO inhibitory activity and their interaction mechanism was studied coupled with molecular simulations. The results were in favor of the hypothesis that the flavonoids of G. hypoleucum might be the active content for gout treatment by inhibiting XO.

  2. Simultaneous analysis of fourteen tertiary amine stimulants in human urine for doping control purposes by liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Lu Jianghai; Wang San; Dong Ying; Wang Xiaobing; Yang Shuming; Zhang Jianli; Deng Jing; Qin Yang; Xu Youxuan; Wu Moutian [National Doping Control Laboratory, China Anti-Doping Agency, 1st Anding Road, ChaoYang District, Beijing 100029 (China); Ouyang Gangfeng, E-mail: cesoygf@mail.sysu.edu.cn [Research Center for Microextraction and Separation Technology, School of Chemistry and Chemical Engineering, Sun Yat-Sen University, Guangzhou 510275 (China)

    2010-01-04

    A method for the simultaneous screening and confirmation of the presence of fourteen tertiary amine stimulants in human urine by gas chromatography-mass spectrometry (GC-MS) in combination with liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed and validated. Solid phase extraction (SPE) and liquid-liquid extraction (LLE) approaches were utilized for the pre-treatment of the urine samples. The study indicated that the capillary temperature played a significant role in the signal abundances of the protonated molecules of cropropamide and crotethamide under positive ion electrospray ionization (ESI) conditions. In addition, comparison studies of two different pre-treatment approaches as well as the two ionization modes were conducted. The LODs of the developed method for all the analytes were lower than the minimum required performance limit (MRPL) as set forth in the World Anti-Doping Agency (WADA) technical document for laboratories. The human urine sample obtained after oral administration of prolintane.HCl was successfully analyzed by the developed method, which demonstrated the applicability and reliability of the method for routine doping control analysis.

  3. Preparation and chromatographic evaluation of a cysteine-bonded zwitterionic hydrophilic interaction liquid chromatography stationary phase.

    Science.gov (United States)

    Shen, Aijin; Guo, Zhimou; Cai, Xiaoming; Xue, Xingya; Liang, Xinmiao

    2012-03-09

    orthogonality between Click TE-Cys and TSK Amide-80 system was 21.4%, i.e. the selectivity was similar but slightly different from each other. The successful separation of protein digests indicated the great potential of Click TE-Cys stationary phase in the separation of complex samples and applicability in two-dimensional liquid chromatography (2D-LC). Copyright © 2011 Elsevier B.V. All rights reserved.

  4. Determination of esomeprazole in rabbit plasma by liquid chromatography-mass spectrometry and its application to a pharmacokinetic study

    OpenAIRE

    Zhang, Meiling; Wang, Xuebao; Wang, Zhiyi; Deng, Mingjie; Xu, Zhisheng; Zheng, Yuancai; Zhang, Yuan; Huang, Xueli

    2012-01-01

    A sensitive and selective liquid chromatography-mass spectrometry (LC–MS) method for determination of esomeprazole in rabbit plasma was developed and validated. After addition of midazolam as internal standard (IS), protein precipitation by acetonitrile was used as sample preparation, and chromatography involved Agilent SB-C18 column (2.1 x 150 mm, 5.0 μm) using 0.1 % formic acid in water and acetonitrile as a mobile phase with gradient elution. Detection involved positive ion mode...

  5. Simultaneous determination of 14 sulfonamides and tetracyclines in biogas plants by liquid-liquid-extraction and liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Spielmeyer, Astrid; Ahlborn, Jenny; Hamscher, Gerd

    2014-04-01

    A new method for the analysis of sulfonamides and tetracyclines in heterogenic biogas plant input samples and fermentation residues is introduced. Veterinary antibiotics are only partially absorbed in the animal gut; therefore, animal manure can contain high loads of these substances. Animal manure is used for biogas generation, so antibiotics can enter the anaerobic fermentation process this way. However, only little is known about the fate of antibiotics within this process, also due to the lack of suitable analytical methods for this complex sample matrix. Therefore, we developed a method for the analysis of ten sulfonamides (sulfachloropyridazine, sulfadiazine, sulfadimethoxine, sulfaguanidine, sulfamerazine, sulfamethazine, sulfamethoxazol, sulfamethoxypyridazin, sulfapyridine, sulfathiazole) and four tetracyclines (chlortetracycline, doxycycline, oxytetracycline, tetracycline) in biogas plant input and output samples, including a single liquid-liquid-extraction step and analysis via liquid chromatography (LC) and triple quadrupole mass spectrometry. The detection limit of this method ranges from 0.01 to 0.08 mg kg(-1). Matrix calibration using antibiotic-free cattle feces and isotopic-labeled internal standards enables quantification of antibiotics in different matrices such as animal manure, dung, or fermenter outputs with recovery rates between 70 and 130 %. This makes the method suitable for investigating the fate of antibiotics in animal manure and fermentation processes. A screening of 15 German biogas plants revealed the presence of several antibiotics up to 9 mg kg(-1) (201 mg kg(-1) dry matter). During the fermentation process, elimination occurs; however, with the exception of chlortetracycline, the antibiotic content remains in the same order of magnitude.

  6. A new method for the determination of benzophenone-UV filters in human serum samples by dispersive liquid-liquid microextraction with liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Vela-Soria, F; Ballesteros, O; Zafra-Gómez, A; Ballesteros, L; Navalón, A

    2014-04-01

    Benzophenone-UV filters (BP-UV filters) are extensively used in cosmetics products to avoid damaging effects of UV radiation. Despite their low toxicity, many research papers indicate that BP-UV filters are weak endocrine disruptors (EDCs). There are clear relationships between BP-UV filters exposure and several health disorders such as carcinogenesis and malformations observed in animals. In the present work, a new sample treatment procedure by dispersive liquid-liquid microextraction (DLLME) is proposed for the extraction of six BPs, namely benzophenone-1 (BP-1), benzophenone-2 (BP-2), benzophenone-3 (BP-3), benzophenone-6 (BP-6), benzophenone-8 (BP-8) and 4-hydroxybenzophenone (4-OH-BP), in human serum samples, followed by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. The method involves an enzymatic treatment to quantify the total content (free plus conjugated species) of BP-UV filters in serum. The extraction parameters were accurately optimized using multivariate optimization approach. Benzophenone-d₁₀ (BP-d₁₀) was used as surrogate. Limits of quantification (LOQs) ranged from 0.4 to 0.9 ng mL(-1) and inter-day precision (evaluated as relative standard deviation) ranged from 1.9% to 13.1%. The method was validated using matrix-matched calibration and a recovery assay. Recovery rates for spiked samples ranged from 97% to 106%, and acceptable linearity was obtained up to concentrations of 40 ng mL(-1). The method was applied to the determination of the target compounds in human serum samples from 20 randomly selected anonymous individuals. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Analysis of biogenic amines in wines by salting-out assisted liquid-liquid extraction and high-performance liquid chromatography with fluorimetric detection.

    Science.gov (United States)

    Ramos, Rui Miguel; Valente, Inês Maria; Rodrigues, José António

    2014-06-01

    Biogenic amines are nitrogenous organic compounds of low molecular weight that are either formed or metabolized in cells of living organisms and can be found in several food products, being produced mainly by amino acid decarboxylation. When ingested in high concentrations they can induce several health problems in humans. In alcoholic beverages, and especially in wine, they are formed during the vinification process as a result of the action of microorganisms. In this work it is proposed a new methodology for the determination of biogenic amines in wines, which includes a sample preparation approach based on salting-out assisted liquid-liquid extraction, the use of dansyl chloride for the derivatization and chromatographic separation by high-performance liquid chromatography with fluorimetric detection. The salting-out effect is used to promote phase separation between water and a water-miscible organic solvent, while improving the extraction of organic or inorganic species. Several extraction parameters were optimized, such as the dansyl chloride concentration, pH and the effects caused by the order in which the extraction and derivatization were performed. Extraction of amines, and consequent detection, depends on the presence of dansyl chloride in solution prior to extraction. The results showed the possibility to simultaneously perform the extraction and the derivatization, making sample preparation easier and less time-consuming. The methodology was successfully applied to the determination of biogenic amines in five wines (white, red and rosé). This method has the potential to be a good alternative to existing methods since it is cheaper, easier and simplifies the sample preparation step. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. An evaluation of cis- and trans-retinol contents in juices using dispersive liquid-liquid microextraction coupled to liquid chromatography with fluorimetric detection.

    Science.gov (United States)

    Viñas, Pilar; Bravo-Bravo, María; López-García, Ignacio; Hernández-Córdoba, Manuel

    2013-01-15

    This study describes a method for coupling dispersive liquid-liquid microextraction (DLLME) and normal-phase liquid chromatography (NP-LC) with fluorescence detection for vitamin A determination with the view to developing a new green sample preparation technique. Parameters affecting DLLME, including the nature and volume of both extractant and disperser solvents, salt addition and time and speed of the centrifugation step, were optimized. The sample was saponified according to European Standards to convert all forms of vitamin A to retinol. For microextraction, 8 mL water were placed in a glass tube with conical bottom and the saponified sample consisting of 2 mL of the methanolic extract containing 100 μL tetrachloroethane was rapidly injected by syringe, thereby forming a cloudy solution. Phase separation was performed by centrifugation, and a volume of 20 μL of the sedimented phase was analyzed by NP-LC. The enrichment factor, calculated as the ratio between the slopes of DLLME-LC and direct LC, was 50 ± 3. The matrix effect was evaluated for different juice samples, and it was concluded that sample quantification can be carried out by aqueous calibration when the standards are also submitted to saponification. The proposed method was applied for determining both cis- and trans-retinol isomers in commercial juices of different types. The intraday and interday precisions were lower than 6% in terms of relative standard deviation. The method was validated using two certified reference materials. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Rapid determination of lipophilic vitamins in human serum by ultra-high performance liquid chromatography using a fluorinated column and high-throughput miniaturized liquid-liquid extraction.

    Science.gov (United States)

    Cervinkova, Barbora; Krcmova, Lenka Kujovska; Klabackova, Sava; Solichova, Dagmar; Solich, Petr

    2017-09-01

    A high-throughput miniaturized liquid-liquid extraction procedure followed by a simple ultra-high performance liquid chromatography method coupled with fluorescence detection for bioanalytical analysis of all tocopherol isomers and retinol in human serum has been developed and validated. In the extraction procedure, a synthetic internal standard tocol was used, which does not occur in the human body. The separation of structurally related vitamins was achieved using a new generation of pentafluorophenyl propyl core-shell stationary phase with elution using methanol and an aqueous solution of ammonium acetate. The fluorescence of retinol and tocopherol isomers was detected at λex  = 325, 295 nm and λem  = 480, 325 nm, respectively. The rapid baseline separation of all analytes was accomplished within 4.0 min. The sensitivity of method was demonstrated with lower limits of quantification: retinol 0.01 μM, α-tocopherol 0.38 μM, β-tocopherol 0.18 μM, γ-tocopherol 0.14 μM, and δ-tocopherol 0.01 μM. Possible application of this method in clinical practice was confirmed by the analysis of human serum samples from healthy volunteers. Finally, the simultaneous determination of retinol and all tocopherol isomers in human serum can enable the clarification of their role in metabolism and in diseases such as cancer. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Method development for the quantitation of ABT-578 in rabbit artery tissue by 96-well liquid-liquid extraction and liquid chromatography/tandem mass spectrometric detection.

    Science.gov (United States)

    Ji, Qin C; Zhang, Jun; Rodila, Ramona; Watson, Pamela; El-Shourbagy, Tawakol

    2004-01-01

    Quantitative determination of drug concentrations in tissue samples can provide critical information for drug metabolism, kinetics, and toxicity evaluations. For analysis of tissue samples using liquid chromatography/tandem mass spectrometric (LC/MS/MS) detection, homogenization is a critical step in achieving good assay performance. Assay performance can be closely evaluated by spiking the drug directly into tissue samples prior to homogenization. It is especially important to include this assay evaluation for the analysis of artery tissue samples because artery tissue is very elastic, making it quite a challenge to develop an effective procedure for homogenization. An LC/MS/MS assay in 96-well format using liquid-liquid extraction was developed for analyzing ABT-578 in rabbit artery samples. Tissue quality control samples were prepared by spiking ABT-578 stock solutions directly into the tissue before homogenization. The usage of the tissue control samples gives a thorough evaluation of the sample preparation process that includes both homogenization and sample extraction. A 20% blood in saline solution was used as a homogenization solution. Calibration standards were made by spiking ABT-578 into rabbit whole blood. Blood quality control samples were also prepared by spiking ABT-578 into rabbit whole blood. These blood QC samples were used to confirm the validity of the calibration curve. A lower limit of quantitation of 0.050 ng/mL was achieved. The linear dynamic range of blood standards was from 0.050-30.3 ng/mL with the correlation coefficient (r) ranging from 0.9969-0.9996. Overall %CV was between 1.3 and 7.0%, and analytical recovery was between 98.2 and 105.8% for blood QC samples. The %CVs for tissue QC samples were between 6.7 and 13.0%, and analytical recovery after correction was between 93.5 and 114.3%. Copyright 2004 John Wiley & Sons, Ltd.

  11. Rapid extraction and determination of amphetamines in human urine samples using dispersive liquid-liquid microextraction and solidification of floating organic drop followed by high performance liquid chromatography.

    Science.gov (United States)

    Ahmadi-Jouibari, Toraj; Fattahi, Nazir; Shamsipur, Mojtaba

    2014-06-01

    A novel, rapid, simple and sensitive dispersive liquid-liquid microextraction method based on the solidification of floating organic drop (DLLME-SFO) combined with high-performance liquid chromatography-ultraviolet detection (HPLC-UV) was used to determine amphetamine and methamphetamine in urine samples. The factors affecting the extraction efficiency of DLLME-SFO such as the kind and volume of the extraction and the disperser solvents, effect of concentration of K2CO3 and extraction time were investigated and the optimal extraction conditions were established. Under the optimum conditions (extraction solvent: 30.0μl 1-undecanol; disperser solvent: 300μl acetonitrile; buffer concentration: 2% (w/v) K2CO3 and extraction time: 1min), calibration curves are linear in the range of 10-3000μgl(-1) and limit of detections (LODs) are in the range of 2-8μgl(-1). The relative standard deviations (RSDs) for 100μgl(-1) of amphetamine and methamphetamine in diluted urine are in the range of 6.2-7.8% (n=7). The method was successfully applied for the determination of amphetamine and methamphetamine in the actual urine samples. The relative recoveries of urine samples spiked with amphetamine and methamphetamine are 87.8-113.2%. The obtained results show that DLLME-SFO combined with HPLC-UV is a fast and simple method for the determination of amphetamine and methamphetamine in urine. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Urinary detection of conjugated and unconjugated anabolic steroids by dilute-and-shoot liquid chromatography-high resolution mass spectrometry.

    Science.gov (United States)

    Tudela, Eva; Deventer, Koen; Geldof, Lore; Van Eenoo, Peter

    2015-02-01

    Anabolic androgenic steroids (AAS) are an important class of doping agents. The metabolism of these substances is generally very extensive and includes phase-I and phase-II pathways. In this work, a comprehensive detection of these metabolites is described using a 2-fold dilution of urine and subsequent analysis by liquid chromatography-high resolution mass spectrometry (LC-HRMS). The method was applied to study 32 different metabolites, excreted free or conjugated (glucuronide or sulfate), which permit the detection of misuse of at least 21 anabolic steroids. The method has been fully validated for 21 target compounds (8 glucuronide, 1 sulfate and 12 free steroids) and 18 out of 21 compounds had detection limits in the range of 1-10 ng mL(-1) in urine. For the conjugated compounds, for which no reference standards are available, metabolites were synthesized in vitro or excretion studies were investigated. The detection limits for these compounds ranged between 0.5 and 18 ng mL(-1) in urine. The simple and straightforward methodology complements the traditional methods based on hydrolysis, liquid-liquid extraction, derivatization and analysis by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS). Copyright © 2014 John Wiley & Sons, Ltd.

  13. Determination of Organic Pollutants in Small Samples of Groundwaters by Liquid-Liquid Extraction and Capillary Gas Chromatography

    DEFF Research Database (Denmark)

    Harrison, I.; Leader, R.U.; Higgo, J.J.W.

    1994-01-01

    A method is presented for the determination of 22 organic compounds in polluted groundwaters. The method includes liquid-liquid extraction of the base/neutral organics from small, alkaline groundwater samples, followed by derivatisation and liquid-liquid extraction of phenolic compounds after neu...

  14. Accelerated solvent extraction combined with dispersive liquid-liquid microextraction before gas chromatography with mass spectrometry for the sensitive determination of phenols in soil samples.

    Science.gov (United States)

    Xing, Han-Zhu; Wang, Xia; Chen, Xiang-Feng; Wang, Ming-Lin; Zhao, Ru-Song

    2015-05-01

    A method combining accelerated solvent extraction with dispersive liquid-liquid microextraction was developed for the first time as a sample pretreatment for the rapid analysis of phenols (including phenol, m-cresol, 2,4-dichlorophenol, and 2,4,6-trichlorophenol) in soil samples. In the accelerated solvent extraction procedure, water was used as an extraction solvent, and phenols were extracted from soil samples into water. The dispersive liquid-liquid microextraction technique was then performed on the obtained aqueous solution. Important accelerated solvent extraction and dispersive liquid-liquid microextraction parameters were investigated and optimized. Under optimized conditions, the new method provided wide linearity (6.1-3080 ng/g), low limits of detection (0.06-1.83 ng/g), and excellent reproducibility (solvent extraction with dispersive liquid-liquid microextraction as a sample pretreatment procedure coupled with gas chromatography and mass spectrometry is an excellent method for the rapid analysis of trace levels of phenols in environmental soil samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Evaluation of aminoacids in irradiated beans (Vigna unguiculata (L.) Walp) by high performance liquid chromatography (HPLC)

    Energy Technology Data Exchange (ETDEWEB)

    Lima, Keila S. Cople; Souza, Luciana B.; Coelho, Maysa J.; Lima, Antonio L. Santos; Hernandes, Nilber K. [Instituto Militar de Engenharia (IME), Rio de Janeiro, RJ (Brazil). Secao de Engenharia Nuclear]. E-mail: keila@ime.eb.br; Godoy, Ronoel L.O. [EMBRAPA Agroindustria de Alimentos, Rio de Janeiro, RJ (Brazil)]. E-mail: ronoel@ctaa.embrapa.br

    2007-07-01

    Fradinho-bean (Vigna unguiculata (L.) Walp) is originated from Africa and is known in Brazil as 'caupi', 'corda' or 'macassar'. It is grown in the interior of Northeast Brazil (semi-arid region) and can be found in parts of the North, being one of the most important components of people's diet in those regions. The Northeast area produces around 429,375 ton of fradinho-bean per year. Leguminous plants are very important sources of proteins, vitamins, carbohydrates and minerals. This kind of bean is an excellent source of proteins (around 23- 25% of its nutritional content), being superior to regular beans (Phaseolus vulgaris). The irradiation process is an alternative to avoid post-harvesting losses, without changing the nutritional value of food. This study has the objective to evaluate the effect of different gamma irradiation doses (0.0; 0.5; 1.0; 2.5; 5.0 and 10.0 kGy) on aminoacid content of fradinho-bean by high performance liquid chromatography (HPLC) and the accompanying of the grains during storage time of 6 months. After irradiation, the bean grains went through a milling process in order to make flour for posterior extraction. A liquid chromatographer Waters, model Alliance 2695, with fluorescent detector Waters 2475, having a mobile phase with gradient elution of sodium acetate. acetonitrile and Milli-Q water, was employed. The flux used was 1 mL/min and the injection volume of 10 {mu}L. The column (C 18 150.0 x 3.9 mm) was kept at 36 deg C. The results show that gamma irradiation is a promise process for fradinho bean during conservation storage time of 6 months, until the dose of 10.0 kGy. Even the most radio-sensitive aminoacids like aromatics and basic lateral chains were preserved. (author)

  16. Cavity ring-down spectroscopy as a detector for liquid chromatography.

    Science.gov (United States)

    Snyder, Kate L; Zare, Richard N

    2003-07-01

    We have demonstrated the use of cavity ring-down spectroscopy (CRDS) as a detector for high performance liquid chromatography (HPLC). For this use, we have designed and implemented a Brewster's angle flow cell such that cavity ring-down spectroscopy can be performed on microliter volumes of liquids. The system exhibits a linear dynamic range of 3 orders of magnitude (30 nM to 30 microM quinalizarin at 470 nm) for static measurements and 2 orders of magnitude (0.5 microM to 50 microM) for HPLC measurements. For the static measurements, the baseline noise is 2.8 x 10(-6) AU rms and 1.0 x 10(-5) AU peak-to-peak, and for the HPLC separations, it is 3.2 x 10(-6) AU rms and 1.3 x 10(-5) AU peak-to-peak. The baseline noise is determined after the data are smoothed by an 11-point boxcar average. The peak areas detected from HPLC separations are reproducible to within 2-3%. The HPLC mass detection limit for a molecule with epsilon = 9 x 10(3) M(-1) cm(-1) in a 300-microm path length cell (illuminated volume, 0.5 microL) is reported as 2.5 x 10(-8) g/mL. These results were obtained using a simple pulsed CRDS system and are comparable to, if not better than, a high-quality commercial UV-vis absorption detector for the same path length.

  17. Analysis of the Constituents in “Zhu She Yong Xue Shuan Tong” by Ultra High Performance Liquid Chromatography with Quadrupole Time-of-Flight Mass Spectrometry Combined with Preparative High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Lin-Lin Wang

    2015-11-01

    Full Text Available “Zhu She Yong Xue Shuan Tong” lyophilized powder (ZSYXST, consists of a series of saponins extracted from Panax notoginseng, which has been widely used in China for the treatment of strokes. In this study, an ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS combined with preparative high performance liquid chromatography (PHPLC method was developed to rapidly identify both major and minor saponins in ZSYXST. Some high content components were removed through PHPLC in order to increase the sensitivity of the trace saponins. Then, specific characteristic fragment ions in both positive and negative mode were utilized to determine the types of aglycone, saccharide, as well as the saccharide chain linkages. As a result, 94 saponins, including 20 pairs of isomers and ten new compounds, which could represent higher than 98% components in ZSYXST, were identified or tentatively identified in commercial ZSYXST samples.

  18. Metabolite Profile of Salidroside in Rats by Ultraperformance Liquid Chromatography Coupled with Quadrupole Time-of-Flight Mass Spectrometry and High-Performance Liquid Chromatography Coupled with Quadrupole-Linear Ion Trap Mass Spectrometry.

    Science.gov (United States)

    Hu, Zhiwei; Wang, Ziming; Liu, Yong; Wu, Yan; Han, Xuejiao; Zheng, Jian; Yan, Xiufeng; Wang, Yang

    2015-10-21

    In the present work, the salidroside metabolite profile in rat urine was investigated, and subsequently the metabolic pathways of salidroside were proposed. After administrations of salidroside at an oral dose of 100 or 500 mg/kg, rat urine samples were collected and pretreated with methanol to precipitate the proteins. The pretreated samples were analyzed by an Acquity ultraperformance liquid chromatography (UPLC) coupled with an HSS T3 column and detected by quadrupole time-of-flight mass spectrometry (Q-TOF-MS) or high-performance liquid chromatography coupled with hybrid triple-quadrupole linear ion trap mass spectrometry (HPLC/Q-trap-MS). A total of eight metabolites were detected and identified on the basis of the characteristics of their protonated ions in the urine samples. The results elucidated that salidroside was metabolized via glucuronidation, sulfation, deglycosylation, hydroxylation, methylation, and dehydroxylation pathways in vivo.

  19. Determination of chlorobenzenes in textiles by pressurized hot water extraction followed by vortex-assisted liquid-liquid microextraction and gas chromatography-mass spectrometry.

    Science.gov (United States)

    Lu, Yang; Zhu, Yan

    2013-12-06

    A method for quantitative determination of chlorobenzenes in textiles is developed, using pressurized hot water extraction (PHWE), vortex-assisted liquid-liquid microextraction (VALLME) and gas chromatography-mass spectrometry (GC-MS). VALLME serves as a trapping step after PHWE. The extraction conditions are investigated, as well as the quantitative features such as linearity, limits of detection (LODs), limits of quantification (LOQs), repeatabilities and reproducibilities between days. LOQs of 0.018-0.032mg/kg were achieved. The present method provides good repeatabilities (RSDtextiles. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Profiling of drug binding proteins by monolithic affinity chromatography in combination with liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Zhang, Xuepei; Wang, Tongdan; Zhang, Hanzhi; Han, Bing; Wang, Lishun; Kang, Jingwu

    2014-09-12

    A new approach for proteome-wide profiling drug binding proteins by using monolithic capillary affinity chromatography in combination with HPLC-MS/MS is reported. Two immunosuppresive drugs, namely FK506 and cyclosporin A, were utilized as the experimental models for proof-of-concept. The monolithic capillary affinity columns were prepared through a single-step copolymerization of the drug derivatives with glycidyl methacrylate and ethylene dimethacrylate. The capillary chromatography with the affinity monolithic column facilitates the purification of the drug binding proteins from the cell lysate. By combining the capillary affinity column purification and the shot-gun proteomic analysis, totally 33 FK506- and 32 CsA-binding proteins including all the literature reported target proteins of these two drugs were identified. Among them, two proteins, namely voltage-dependent anion-selective channel protein 1 and serine/threonine-protein phosphatase PGAM5 were verified by using the recombinant proteins. The result supports that the monolithic capillary affinity chromatography is likely to become a valuable tool for profiling of binding proteins of small molecular drugs as well as bioactive compounds. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Comprehensive analysis of umami compounds by ion-pair liquid chromatography coupled to mass spectrometry.

    Science.gov (United States)

    Coulier, Leon; Bas, Richard; Hekman, Maarten; van der Werff, Bianca J C; Burgering, Maurits; Thissen, Uwe

    2011-09-01

    An ion-pair LC-ESI-MS method was developed capable of analyzing various reported umami or umami-enhancing compounds, including glutamic acid and 5'-ribonucleotides. The method was validated using tomato and potato samples and showed overall good analytical performance with respect to selectivity, detection limit, linearity, and repeatability. The method was applied to various tomato samples resulting in concentrations of glutamic acid and 5'-ribonucleotides that were in good comparison with literature. The methodology might also be used for the discovery of new umami (enhancing) compounds in an untargeted mode. This was to a certain extent demonstrated for tomato samples by correlating all peaks observed with the ion-pair liquid chromatography-mass spectrometry (LC-MS) method to sensory properties using multivariate statistics. This study describes the development and application of a LC-MS method, which can be used to quantify several known umami (enhancing) compounds in various foods. Furthermore, the method might be useful for the discovery of new umami (enhancing) compounds. © 2011 Institute of Food Technologists®

  2. [Determination of three isothiazolinone biocides in water-borne adhesives by high performance liquid chromatography].

    Science.gov (United States)

    Zhou, Xiao; Li, Xiaolan; Chen, Zhiyan; Ye, Changwen; Zhou, Yun; Meng, Dongling

    2015-01-01

    A rapid high performance liquid chromatography (HPLC) method was developed for the quantitative analysis of three isothiazolinone biocides (2-methyl-4-isothiazolin-3-one (MI), 5-chloro-2-methyl-4-isothiazolin-3-one (CMI) and 1, 2-benzylisothiazolin-3-one (BIT)) in water-borne adhesives. The sample was extracted with methanol-water (1:1, v/v), and purified by centrifugation and filtration. The isothiazolones were separated on a C18 column with methanol-water as mobile phases under gradient elution and detected with a diode array detector (DAD). The pretreatment factors such as extraction solvent, extraction method, dilution ratio, extraction time were optimized. Under the optimized conditions, the targets had good linearities (r2H > or = 0.9992) in the range of 0.25-10.0 mg/L. The recoveries were between 92% and 103% with the relative standard deviations not more than 4%. The limits of detection (LODs) were between 0.43 mg/kg and 1.14 mg/kg. The limits of quantification (LOQs) were between 1.44 mg/kg and 3.81 mg/kg. The results showed that the method can achieve the purpose of quantitative detection. The analyses of real samples verified the reliability of this method.

  3. Simultaneous quantification of five major biologically active ingredients of saffron by high-performance liquid chromatography.

    Science.gov (United States)

    Li, N; Lin, G; Kwan, Y W; Min, Z D

    1999-07-23

    A simple, sensitive and specific high-performance liquid chromatography-UV (HPLC-UV) method has been developed for the first time to simultaneously quantify the five major biologically active ingredients of saffron, namely crocin 1, crocin 2, crocin 3, crocin 4 and crocetin. Calibration curves were derived by spiking authentic compounds and internal standard, 13-cis-retinoic acid, into herbal samples prior to extraction. Extraction was conducted simply by stirring dried herb (20 mg) with 80% aqueous methanol (5 ml) at ambient temperature in the dark for 2 h. The HPLC assay was performed on a reversed-phase C18 column with linear gradient elution using methanol and 1% aqueous acetic acid. Calibrations were linear (r2 = 0.999) for all five analytes, with overall intra- and inter-day RSDs of less than 11%. The assay was successfully applied to the determination of four crocins and crocetin in three saffron samples and two Zhizi, another crocin-containing herb. Results indicate that the developed HPLC assay can be readily utilized as a quality control method for crocin-containing medicinal herbs.

  4. Rapid Preconcentration for Liquid Chromatography-Mass Spectrometry Assay of Trace Level Neuropeptides

    Science.gov (United States)

    Zhou, Ying; Mabrouk, Omar S.; Kennedy, Robert T.

    2013-11-01

    Measurement of neuropeptides in the brain through in vivo microdialysis sampling provides direct correlation between neuropeptide concentration and brain function. Capillary liquid chromatography-multistage mass spectrometry (CLC-MSn) has proven to be effective at measuring endogenous neuropeptides in microdialysis samples. In the method, microliter samples are concentrated onto nanoliter volume packed beds before ionization and mass spectrometry analysis. The long times required for extensive preconcentration present a barrier to routine use because of the many samples that must be analyzed and instability of neuropeptides. In this study, we evaluated the capacity of 75 μm inner diameter (i.d.) capillary column packed with 10 μm reversed phase particles for increasing the throughput in CLC-MSn based neuropeptide measurement. Coupling a high injection flow rate for fast sample loading/desalting with a low elution flow rate to maintain detection sensitivity, this column has reduced analysis time from ˜30 min to 3.8 min for 5 μL sample, with 3 pM limit of detection (LOD) for enkephalins and 10 pM LOD for dynorphin A1-8 in 5 μL sample. The use of isotope-labeled internal standard lowered peptide signal variation to less than 5 %. This method was validated for in vivo detection of Leu and Met enkephalin with microdialysate collected from rat globus pallidus. The improvement in speed and stability makes CLC-MSn measurement of neuropeptides in vivo more practical.

  5. Current role of liquid chromatography-mass spectrometry in clinical and forensic toxicology.

    Science.gov (United States)

    Maurer, Hans H

    2007-08-01

    This paper reviews multi-analyte single-stage and tandem liquid chromatography-mass spectrometry (LC-MS) procedures using different mass analyzers (quadrupole, ion trap, time-of-flight) for screening, identification, and/or quantification of drugs, poisons, and/or their metabolites in blood, plasma, serum, or urine published after 2004. Basic information about the biosample assayed, work-up, LC column, mobile phase, ionization type, mass spectral detection mode, and validation data of each procedure is summarized in tables. The following analytes are covered: drugs of abuse, analgesics, opioids, sedative-hypnotics, benzodiazepines, antidepressants including selective-serotonin reuptake inhibitors (SSRIs), herbal phenalkylamines (ephedrines), oral antidiabetics, antiarrhythmics and other cardiovascular drugs, antiretroviral drugs, toxic alkaloids, quaternary ammonium drugs and herbicides, and dialkylphosphate pesticides. The pros and cons of the reviewed procedures are critically discussed, particularly, the need for studies on matrix effects, selectivity, analyte stability, and the use of stable-isotope labeled internal standards instead of unlabeled therapeutic drugs. In conclusion, LC-MS will probably become a gold standard for detection of very low concentrations particularly in alternative matrices and for quantification in clinical and forensic toxicology. However, some drawbacks still need to be addressed and finally overcome.

  6. Sensitive quantification of diphemanil methyl sulphate in human plasma by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Do, B; Goulay-Dufaÿ, S; Le Hoang, M D; Adoui, N; Graffard, H; Guyon, F; Pradeau, D

    2007-01-01

    A simple detection system with a high-performance liquid chromatography (HPLC) with positive ionisation-tandem mass spectrometry (ESI-MS/MS) for determining diphemanil methylsulphate (DMS) levels in human plasma using 4-diphemanylmethylene,1-methylpiperidine as an internal standard (I.S.), is proposed. The acquisition was performed with the multiple reactional monitoring (MRM) mode, by monitoring the transitions: m/z 278>262 for DMS and m/z 263>247 for the I.S. The method involved a simple single-step deproteinisation with acetonitrile. The analyte was chromatographed on a Zorbax C18 reversed-phase chromatographic column by isocratic elution with 10(-3)M ammonium acetate and 10(-3)M hexafluorobutyric acid, adjusted to pH 7.0 with ammoniac/acetonitrile (40/60, v/v). The results were linear over the studied range (0.5-50.0 ng mL(-1)) and the total analysis time for each run was 10 min. The mean extraction apparent recoveries expressed at the 95% intervals of confidence were 94-104% for DMS and 92-106% for the I.S. The intra- and inter-assay precisions were 4.6-8.4% and 2.9-10.6%, respectively. The limit of quantification was 0.15 ng mL(-1). The devised assay was successfully applied to the residual concentrations monitoring in infant.

  7. Liquid chromatography-tandem mass spectrometry for analysis of intestinal permeability of loperamide in physiological buffer.

    Directory of Open Access Journals (Sweden)

    Miriam S Rubelt

    Full Text Available Analysis of in vitro samples with high salt concentrations represents a major challenge for fast and specific quantification with liquid chromatography-tandem mass spectrometry (LC-MS/MS. To investigate the intestinal permeability of opioids in vitro employing the Ussing chamber technique, we developed and validated a fast, sensitive and selective method based on LC-MS/MS for the determination of loperamide in HEPES-buffered Ringer's solution. Chromatographic separation was achieved with an Atlantis dC18 column, 2.1 mm×20 mm, 3 µm particle size and a gradient consisting of methanol/0.1% formic acid and ammonium acetate. The flow rate was 0.7 ml/min, and the total run time was 3 min. For quantification, two mass transitions for loperamide and a deuterated internal standard (methadone-d(3 were used. The lower limit of loperamide quantification was 0.2 ng/ml. This new LC-MS/MS method can be used for the detection of loperamide in any experimental setup using HEPES-buffered Ringer's solution as a matrix compound.

  8. Liquid chromatography quadrupole time-of-flight mass spectrometry selective determination of ochratoxin A in wine.

    Science.gov (United States)

    Rodríguez-Cabo, T; Rodríguez, I; Ramil, M; Cela, R

    2016-05-15

    The performance of liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) for ochratoxin A (OTA) determination in wine is evaluated for the first time. Sample preparation was optimized to obtain quantitative recoveries at the same time that the efficiency of electrospray ionization (ESI) remained unaltered between sample extracts and calibration standards. Under final conditions, samples (20 mL) were concentrated using a reversed-phase solid-phase extraction (SPE) cartridge, followed by OTA elution with 1 mL of ethyl acetate. The absolute recoveries of the method, established against calibration standards, were 91-121% and 90-113% (without and with internal standard correction, respectively), for wines fortified at 3 concentration levels. The attained LOQ (0.05 ng mL(-1)) remained below the maximum permitted OTA concentration (2 ng mL(-1)) in dry wines. The method was applied to different samples, with OTA being found in some dessert wines at concentrations below 1 ng mL(-1). The ethyl ester of OTA (OTC) could be identified in the same wine samples from its accurate full product ion spectra. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Optimization of solid phase microextraction coatings for liquid chromatography mass spectrometry determination of neurotransmitters.

    Science.gov (United States)

    Cudjoe, Erasmus; Pawliszyn, Janusz

    2014-05-09

    A simple solid phase microextraction method coupled to liquid chromatography mass spectrometry is introduced for the analysis of neurotransmitter compounds with a wide range of polarities in biological matrices. A novel "reversed" reverse-phase chromatographic method was developed without pre-column derivatization for the analysis of dopamine, serotonin, gamma aminobutyric acid and glutamate. New solid phase microextraction "in house" coatings using mixed-mode solid phase extraction particles were prepared, and used for the extraction of polar neurotransmitters. The polymer-support base reverse phase mixed-mode sorbents with strong ion exchange properties generally had higher extraction efficiencies compared to similar sorbents with weak ion exchange properties. The linear range was determined to be between 0.01 and 150ng/mL for all the analytes, except for GABA, which was from 0.1 to 100ng/mL. The limit of detection range was from 6 to 10pg/mL for all the neurotransmitters, and the limits of quantitation were in the range of 20-35pg/mL. The results demonstrate the potential of the SPME-LC-MS/MS technique for bioanalysis of small polar endogenous compounds, such as neurotransmitters, from various biological matrices using the mixed-mode sorbents as the extraction phase. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Multiplicative effects model with internal standard in mobile phase for quantitative liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Song, Mi; Chen, Zeng-Ping; Chen, Yao; Jin, Jing-Wen

    2014-07-01

    Liquid chromatography-mass spectrometry assays suffer from signal instability caused by the gradual fouling of the ion source, vacuum instability, aging of the ion multiplier, etc. To address this issue, in this contribution, an internal standard was added into the mobile phase. The internal standard was therefore ionized and detected together with the analytes of interest by the mass spectrometer to ensure that variations in measurement conditions and/or instrument have similar effects on the signal contributions of both the analytes of interest and the internal standard. Subsequently, based on the unique strategy of adding internal standard in mobile phase, a multiplicative effects model was developed for quantitative LC-MS assays and tested on a proof of concept model system: the determination of amino acids in water by LC-MS. The experimental results demonstrated that the proposed method could efficiently mitigate the detrimental effects of continuous signal variation, and achieved quantitative results with average relative predictive error values in the range of 8.0-15.0%, which were much more accurate than the corresponding results of conventional internal standard method based on the peak height ratio and partial least squares method (their average relative predictive error values were as high as 66.3% and 64.8%, respectively). Therefore, it is expected that the proposed method can be developed and extended in quantitative LC-MS analysis of more complex systems. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Determination of levofloxacin in human serum using liquid chromatography tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Samiksha Ghimire

    2018-01-01

    Full Text Available A rapid liquid chromatography tandem-mass spectrometry method was developed for the determination of levofloxacin and its metabolite (desmethyl-levofloxacin in human serum. Sample preparation was done using protein precipitation technique. Our method had a run time of 2.5 min and retention times of 1.6 min for all analytes. The standard curves were linear within the concentration range of 0.10 to 5.00 mg/L for levofloxacin and 0.10 to 4.99 mg/L for desmethyl- levofloxacin; a correlation coefficient (R2 of 0.999 and 0.998 respectively. The lower limit of quantification for both analytes was 0.10 mg/L. Within-day precision ranged from 1.4% and 2.4% for levofloxacin, 1.5% to 5% for desmethyl-levofloxacin and between-day precision ranged from 3.6% to 4.1% for levofloxacin and 0.0% to 3.3% for desmethyl-levofloxacin; whereas, accuracy ranged from 0.1% to 12.7% for levofloxacin and 0.2% to 15.6% for desmethyl-levofloxacin. This method could be a useful asset for routine therapeutic drug monitoring of levofloxacin in multi-drug resistant tuberculosis patients.

  12. Determination of meropenem levels in human serum by high-performance liquid chromatography with ultraviolet detection.

    Science.gov (United States)

    Roth, Thomas; Fiedler, Stella; Mihai, Sidonia; Parsch, Hans

    2017-05-01

    Meropenem is a β-lactam broad-spectrum antibiotic and belongs to the subgroup of carbapenems. It is primarily used in intensive care units for intravenous treatment of severe infections. To avoid bacterial resistance or toxic side effects, the determination of serum meropenem concentration is highly advisable. A simple and fast method for the quantitative determination of meropenem in human serum using high-performance liquid chromatography with ultraviolet detection (HPLC/UV) was developed and validated. Meropenem was determined by an isocratic HPLC using a tris(hydroxymethyl)aminomethane buffer (pH 8.5; 15% methanol) as a mobile phase and UV detection at 300 nm, with a flow rate of 1.0 mL/min and an analysis time of 10 min. Chromatographic separation was performed on a Kinetex C18 column (5 μm, 150 × 4.6 mm). In order to remove undesired serum components, solid-phase extraction was used for sample preparation. Since meropenem is not stable in solution, sample and stock solution were stored at -80°C. After preparation, samples were stable at room temperature for at least 6 h. The calibration curve was linear from 3.5 to 200 mg/L with a correlation coefficient r2 of 0.999. The method is accurate with an intra- and inter-assay precision <18.5%. Copyright © 2016 John Wiley & Sons, Ltd.

  13. Determination of mono- and dichloroacetic acids in betaine media by liquid chromatography.

    Science.gov (United States)

    Ghassempour, Alireza; Chalavi, Soheila; Abdollahpour, Assem; Mirkhani, Seyed Amir

    2006-02-15

    A simple and sensitive method has been developed for the analysis of residue amounts of chloroacetic acids in betaine samples based on derivatization by 1-naphthylamine (NA). The derivatized compounds are analyzed by reverse phase high performance liquid chromatography using methanol and water as mobile phase in the ratio of 32/68 (v/v) and phenyl column and PDA detection at 222nm. The detection limits (LOD) of monochloroacetic acid (MCA) and dichloroacetic acid (DCA) are 0.1 and 0.15mugmL(-1), respectively. The limits of quantification (LOQ) and the linear dynamic ranges (LDR) of MCA are found to be 1 and 1-400mugmL(-1), respectively, and for DCA are found to be 3 and 3-400mugmL(-1), respectively. The precision at the 5ppm level for MCA and DCA are about 3% and 2%, (n=5), respectively. The average recovery for MCA and DCA spiked to betaine samples are 98% and 97%, respectively.

  14. Determination of streptomycin and dihydrostreptomycin in milk and honey by liquid chromatography with tandem mass spectrometry.

    Science.gov (United States)

    van Bruijnsvoort, Michel; Ottink, Stef J M; Jonker, Klaas M; de Boer, Enne

    2004-11-26

    Two liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were developed for the determination of streptomycin (STR) and its derivative dihydrostreptomycin (DHSTR) in milk and honey. These aminoglycoside antibiotics are used as veterinary drugs. In the EU, the presence of dihydro- and streptomycin residues in honey is forbidden, the maximum residue level (MRL) in milk is 200 microg/kg. The methods were optimised with regard to sensitivity and chromatographic efficiency, and validated by a procedure consistent with EU directive 2002/657. Average recoveries and accompanying standard deviations were satisfactory. The limit of quantification of STR was 2 microg/kg in honey and 10 microg/kg in milk, of DHSTR it was a factor two lower. The precision of the milk analysis was improved by using STR as the internal standard for DHSTR and vice versa. In a survey of 186 honeys available on the Dutch market, 26% of the honeys of foreign origin were positive for (DH)STR. This occurence rate was consistent with previous surveys, but lower concentrations were found.

  15. Comparative determination of phenytoin in plasma by fluorescence polarization immunoassay and high performance liquid chromatography.

    Science.gov (United States)

    Othman, S; al-Turk, W A; Awidi, A S; Daradkeh, T K; Shaheen, O

    1987-09-01

    The need for careful monitoring of plasma concentrations of phenytoin during use of the drug in the treatment of epilepsy is well recognized; there can be great intersubject variation in the absorption rate and clearance rate of the drug, and its therapeutic ratio is narrow. In this study, two methods for determining plasma phenytoin concentrations were compared. One, based on fluorescence polarisation immunoassay (FPIA), is utilised in a commercially-available kit. The other, our own modification of a published procedure, was based on high performance liquid chromatography (HPLC). The accuracy and precision of both methods were evaluated, and the coefficients of variation (C.V.) were calculated. The C.V.'s ranged from 0.71 to 1.86% for the FPIA method, and from 2.81 to 8.69% for the HPLC method. Corresponding bias values were 1.20 to 1.60%, and 2.81 to 8.69%, respectively. A good correlation coefficient (0.977) was obtained, but estimated phenytoin concentrations were significantly higher (95% confidence level) using the HPLC method. We conclude that both methods perform adequately for clinical purposes. The HPLC method is, however, less expensive than the FPIA method.

  16. Quantitative analysis of the tumor suppressor dendrogenin A using liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Noguer, Emmanuel; Soules, Régis; Netter, Claude; Nagarathinam, Citra; Leignadier, Julie; Huc-Claustre, Emilie; Serhan, Nizar; Rives, Arnaud; de Medina, Philippe; Silvente-Poirot, Sandrine; Poirot, Marc

    2017-10-01

    Dendrogenin A (DDA) was recently identified as a mammalian cholesterol metabolite that displays tumor suppressor and neurostimulating properties at low doses. In breast tumors, DDA levels were found to be decreased compared to normal tissues, evidencing a metabolic deregulation of DDA production in cancers. DDA is an amino-oxysterol that contains three protonatable nitrogen atoms. This makes it physico-chemically different from other oxysterols and it therefore requires specific analytical methods We have previously used a two-step method for the quantification of DDA in biological samples: 1) DDA purification from a Bligh and Dyer extract by RP-HPLC using a 250×4.6mm column, followed by 2) nano-electrospray ionization mass spectrometry (MS) fragmentation to analyze the HPLC fraction of interest. We report here the development a liquid chromatography tandem mass spectrometry method for the analysis of DDA and its analogues. This new method is fast (10min), resolving (peak width <4s) and has a weak carryover (<0.01%). We show that this technique efficiently separates DDA from its C17 isomer and other steroidal alkaloids from the same family establishing a proof of concept for the analysis of this family of amino-oxysterols. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Quantitative analysis of racecadotril metabolite in human plasma using a liquid chromatography/tandem mass spectrometry.

    Science.gov (United States)

    Xu, Yu; Huang, Jinchang; Liu, Fei; Gao, Shu; Guo, Qingxiang

    2007-06-01

    Orally administered racecadotril is rapidly hydrolyzed to the more potent enkephalinase inhibitor thiorphan in vivo. A sensitive and specific liquid chromatography/tandem mass spectrometry method was developed and validated to quantify thiorphan in human plasma using lisinopril as the internal standard. After a simple protein precipitation with methanol, the post-treatment samples were analyzed on a CN column interfaced with a triple-quadruple tandem mass spectrometer using negative electrospray ionization. The method was validated to demonstrate the specificity, lower limit of quantification, accuracy, and precision of measurements. The assay was linear over the concentration range 9.38-600 ng/mL using a 5 microL aliquot of plasma. The correlation coefficients for the calibration curves ranged from 0.9985 to 0.9995. The intra- and inter-day precisions over the entire concentration were not more than 6.33%. Methanol and water (35:65, v/v) is used as the isocratic mobile phase, with 0.1% of formic acid in water. The method was successfully applied for pharmacokinetic study after a single oral administration of 200 mg racecadotril to 20 healthy volunteers.

  18. Trace analysis of urea nitrate by liquid chromatography-UV/fluorescence.

    Science.gov (United States)

    de Perre, Chloé; McCord, Bruce

    2011-09-10

    In this paper we have adapted a technique, previously used to determine the presence of urea in aqueous samples of wine and urine, to detect trace levels of urea nitrate explosives. The procedure involves the reaction of the uronium ion (protonated urea) with a fluorophore, xanthydrol. By modification of the procedure to utilize non-aqueous reagents, in neutral conditions, it can be made specific to the presence of the urea nitrate ion pair. The procedure includes selective detection of derivatization products by UV and fluorescence following separation by High-Performance Liquid Chromatography (HPLC). Analytical method development included optimization of HPLC conditions (solvent, gradient), UV and fluorescence wavelengths, and derivatization parameters (xanthydrol amount, reaction times, temperature). The extraction of urea nitrate from surfaces was also investigated and optimized. For best quantification, it was shown that an internal standard was required; this resulted in a quantification limit around 0.17mM (21mg/L). The entire procedure could be performed in less than 30min per sample and potential interferences such as ammonium, nitrate, and urea did not produce a response under standard conditions. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  19. Hard cap espresso extraction and liquid chromatography determination of bioactive compounds in vegetables and spices.

    Science.gov (United States)

    Martinez-Sena, María Teresa; de la Guardia, Miguel; Esteve-Turrillas, Francesc A; Armenta, Sergio

    2017-12-15

    A new analytical procedure, based on liquid chromatography with diode array and fluorescence detection, has been proposed for the determination of bioactive compounds in vegetables and spices after hard cap espresso extraction. This novel extraction system has been tested for the determination of capsaicin and dihydrocapsaicin from fresh chilli and sweet pepper, piperine from ground pepper, curcumin from turmeric and curry, and myristicin from nutmeg. Extraction efficiency was evaluated by using acetonitrile:water and ethanol:water mixtures. The proposed method allows the extraction of samples with 100mL of 60% (v/v) ethanol in water. The obtained limits of quantification for the proposed procedure ranged from 0.07 to 0.30mgg-1 and results were statistically comparable with those obtained by ultrasound assisted extraction. Hard cap espresso machines offer a fast, effective and quantitative tool for the extraction of bioactive compounds from food samples with an extraction time lower than 30s, using a global available and low cost equipment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Rapid screening and characterisation of antioxidants of Cosmos caudatus using liquid chromatography coupled with mass spectrometry.

    Science.gov (United States)

    Shui, Guanghou; Leong, Lai Peng; Wong, Shih Peng

    2005-11-15

    Ulam raja (Cosmos caudatus) is used traditionally for improving blood circulation. In this study, it was found that ulam raja had extremely high antioxidant capacity of about 2,400 mg l-ascorbic acid equivalent antioxidant capacity (AEAC) per 100 g of fresh sample. Antioxidant peaks in extract of ulam raja were firstly characterized using free radical spiking test through high performance liquid chromatography coupled with mass spectrometry (MS). Upon reaction with 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals, intensities of antioxidant peaks will be significantly reduced. HPLC/MS(n) was further applied to elucidate the chemical structures of antioxidant peaks characterized in the spiking test. More than twenty antioxidants were identified in ulam raja, and their chemical structures were proposed. The major antioxidants in ulam raja were attributed to a number of proanthocyanidins that existed as dimers through hexamers, quercetin glycosides, chlorogenic, neo-chlorogenic, crypto-chlorogenic acid and (+)-catching. High content of antioxidants antioxidants contained in ulam raja could be partly responsible for its ability to reduce oxidative stress.

  1. Determination of Decabrominated Diphenyl Ether in Soils by Soxhlet Extraction and High Performance Liquid Chromatography

    Science.gov (United States)

    Yang, Xing-Jian; Dang, Zhi; Zhang, Fang-Li; Lin, Zhao-Ying; Zou, Meng-Yao; Tao, Xue-Qin; Lu, Gui-Ning

    2013-01-01

    This study described the development of a method based on soxhlet extraction combining high performance liquid chromatography (soxhlet-HPLC) for the accurate detection of BDE-209 in soils. The solvent effect of working standard solutions in HPLC was discussed. Results showed that 1 : 1 of methanol and acetone was the optimal condition which could totally dissolve the BDE-209 in environmental samples and avoid the decrease of the peak area and the peak deformation difference of BDE-209 in HPLC. The preliminary experiment was conducted on the configured grassland (1 μg/g) to validate the method feasibility. The method produced reliable reproducibility, simulated soils (n = 4) RSD 1.0%, and was further verified by the analysis e-waste contaminated soils, RSD range 5.9–11.4%. The contamination level of BDE-209 in burning site was consistent with the previous study of Longtang town but lower than Guiyu town, and higher concentration of BDE-209 in paddy field mainly resulted from the long-standing disassembling area nearby. This accurate and fast method was successfully developed to extract and analyze BDE-209 in soil samples, showing its potential use for replacing GC to determinate BDE-209 in soil samples. PMID:24302876

  2. [Serum metabolomics analysis on benign prostate hyperplasia in mice based on liquid chromatography-mass spectrometry].

    Science.gov (United States)

    Geng, Yue; Sun, Fengxia; Ma, Yu; Deng, Ligang; Lü, Jianyun; Li, Teng; Wang, Congcong

    2014-12-01

    Benign prostatic hyperplasia (BPH) increasingly becomes a common factor affecting the quality of life of aging men. Its pathogenesis has not yet been fully elucidated. Ultra-high pressure liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) was employed to detect the changes of serum metabolites in normal mice, benign prostatic hyperplasia model mice and BPH model mice with finasteride intervention. The serum metabolite profiles of the three groups of mice were analyzed. Partial least squares-discriminant analysis (PLS-DA) was used for group differentiation and biomarker selection. The results showed good distinction among the three groups of mice serum metabolite spectra. Three potential biomarkers, 1-hexadecanoyl-SN-glycero-3-phosphocholine, 1-O-hexadecyl-2-O-acetyl-sn-glyceryl-3-phosphorylcholine and (Z)-13-docosenamide, were discovered and identified. They all indicated the occurrence of benign prostatic hypertrophy is closely related to the disorders of lipid metabolism. Coinpared with the control group, the contents of the first two substances were significantly increased in the serum of BPH model mice, and significantly decreased after intervened by finasteride. The contents of (Z)-13-docosenamide decreased significantly in the serum of model group, and increased after intervened by finasteride. Compared with the control group, the contents of three biomarkers in finasteride group did not recover completely and had significant differences. This study is conductive to open new avenues of diagnosis and medical treatment for BPH.

  3. Hydrophilic interaction liquid chromatography method for measuring the composition of aquatic humic substances

    KAUST Repository

    Wang, Renqi

    2015-01-01

    A hydrophilic interaction liquid chromatography (HILIC) method was developed to measure the composition of humic substances from river, reservoir, and treated wastewater based on their physicochemical properties. The current method fractionates the humic substances into four well-defined groups based on parallel analyses with a neutral and a cationic HILIC column, using mobile phases of varied compositions and pH. The results indicate that: (i) the proportion of carboxylic acids in the humic substances from terrestrial origins is less than half of that from treated wastewater (Jeddah, KSA), (ii) a higher content of basic compounds was observed in the humic substances from treated wastewater and Ribou Reservoir (Cholet, France) than in the sample from Loire River (France), (iii) a higher percentage of hydrophobic macromolecules were found in the humic substances from Loire River than in the other samples, and (iv) humic substances of treated wastewater contained less ionic neutral compounds (i.e., pKa 5-9) than the waters from terrestrial origins. The physicochemical property disparity amongst the compounds in each humic substances sample was also evaluated. The humic substances from the lightly humic Loire river displayed the highest disparity, whereas the highly humic Suwannee river (Georgia, USA) showed the most homogeneous humic substances.

  4. Gluten Detection and Speciation by Liquid Chromatography Mass Spectrometry (LC-MS/MS

    Directory of Open Access Journals (Sweden)

    Stephen Lock

    2013-12-01

    Full Text Available Liquid chromatography tandem mass spectrometry (LC-MS/MS has been used historically in proteomics research for over 20 years. However, until recently LC-MS/MS has only been routinely used in food testing for small molecule contaminant detection, for example pesticide and veterinary residue detection, and not as a replacement of microbiological food testing methods, specifically allergen analysis. Over the last couple of years, articles have started to be published which describe the detection of allergens by LC-MS/MS. In this article we will describe how LC-MS/MS can be applied in the area of gluten detection and how it can be used to specifically differentiate the species of gluten used in food, where specific markers for each variety of gluten can be simultaneously acquired and detected at the same time. The article will discuss the effect of variety on the peptide response observed from different wheat grain varieties and will describe the sample preparation protocol which is essential for generating the peptide markers used for speciation.

  5. Is high pressure liquid chromatography an effective screening tool for characterization of molecular defects in hemoglobinopathies?

    Science.gov (United States)

    Moorchung, Nikhil; Phillip, Joseph; Sarkar, Ravi Shankar; Prasad, Rupesh; Dutta, Vibha

    2013-01-01

    Hemoglobinopathies constitute entities that are generated by either abnormal hemoglobin or thalassemias. high pressure liquid chromatography (HPLC) is one of the best methods for screening and detection of various hemoglobinopathies but it has intrinsic interpretive problems. The study was designed to evaluate the different mutations seen in cases of hemoglobinopathies and compare the same with screening tests. 68 patients of hemoglobinopathies were screened by HPLC. Mutation studies in the beta globin gene was performed using the polymerase chain reaction (PCR)-based allele-specific Amplification Refractory Mutation System (ARMS). Molecular analysis for the sickle cell mutation was done by standard methods. The IVS 1/5 mutation was the commonest mutation seen and it was seen in 26 (38.23%) of the cases. This was followed by the IVS 1/1, codon 41/42, codon 8/9, del 22 mutation, codon 15 mutation and the -619 bp deletion. No mutation was seen in eight cases. There was a 100% concordance between the sickle cell trait as diagnosed by HPLC and genetic testing. Our study underlies the importance of molecular testing in all cases of hemoglobinopathies. Although HPLC is a useful screening tool, molecular testing is very useful in accurately diagnosing the mutations. Molecular testing is especially applicable in cases with an abnormal hemoglobin (HbD, HbE and HbS) because there may be a concomitant inheritance of a beta thalassemia mutation. Molecular testing is the gold standard when it comes to the diagnosis of hemoglobinopathies.

  6. Determination of acesulfame and sucralose in oral electrolyte maintenance solution by liquid chromatography.

    Science.gov (United States)

    Johns, Paul; Dowlati, Lobat

    2003-01-01

    A method was developed for the direct, simultaneous determination of acesulfame and sucralose in oral electrolyte maintenance solution (OEMS). Analyte separation and quantitation were achieved by gradient reversed-phase liquid chromatography (LC) and UV absorbance at 192 nm, respectively. Detection at a second wavelength, 214 nm, was used to check sucralose peak purity; 20 microL OEMS was injected without preparation or dilution. System linearity was demonstrated as 192 nm peak area versus analyte concentration at 80-120% OEMS sweetener fortification (r > 0.999, and all residuals sucralose). Spike recoveries for OEMS samples prepared at 3 spiking levels (80, 100, and 120% sweetener fortification) ranged from 100.3 to 102.0% for acesulfame, and from 97.9 to 102.3% for sucralose. In a second assessment of method accuracy, the same spiked OEMS samples were tested by 2 alternative methods: acesulfame (LC/UV at 230 nm) and sucralose (anion exchange-pulsed amperometric detection). Results for the alternative acesulfame method were within 1.2%, and for the alternative sucralose method within 6.0%, of the corresponding results obtained by the 192 nm method. Repeatability and intermediate precision RSD values were sucralose. The limits of quantitation were 1.6 and 32 mg/L for acesulfame potassium and sucralose, respectively. Despite the weak UV absorptivity of sucralose and the consequent small size of its LC peak, no evidence was found for sucralose interference in any of the commercial OEMS flavors.

  7. Multi-detection of preservatives in cheeses by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Fuselli, Fabio; Guarino, Chiara; La Mantia, Alessandro; Longo, Lucia; Faberi, Angelo; Marianella, Rosa Maria

    2012-10-01

    The incorrect use of preservatives in cheeses may compromise food safety and damage consumers. According to the law, more than one preservative may be contemporarily used in cheeses. So a method for their contemporary detection may be useful for both manufacturers and control agencies quality control. In this research a liquid chromatography-tandem mass spectrometric with electrospray ionization method for the multi-determination of seven preservatives (benzoic acid, citric acid, hexamethylenetetramine, lysozyme, natamycin, nisin and sorbic acid) in cheese was developed. The preservatives were contemporarily extracted from cheese by a single procedure, and analyzed by RP-LC/ESI-MS/MS (Ion Trap) in positive ionization mode, with single reaction monitoring (SRM) acquisition. Three sample types (hard, pasta filata and fresh cheese) were used for method evaluation. Recoveries were mostly higher than 90%; MDLs ranged from 0.02 to 0.26 mgkg(-1), and MQLs were included between 0.07 and 0.88 mgkg(-1). Due to matrix effect, quantitation was performed by referring to a matrix matched calibration curve, for each cheese typology. This method was also applied to commercial cheese samples, with good results. It appears fast, reliable and suitable for both screening and confirmation of the presence and quantitation of the preservatives in a single, multi-detection analysis. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC

    Directory of Open Access Journals (Sweden)

    Cheng Bai

    2007-01-01

    Full Text Available High-performance liquid chromatography (HPLC analysis was used for identification of two problematic ureides, asparagine and citrulline. We report here a technique that takes advantage of the predictable delay in retention time of the co-asparagine/citrulline peak to enable both qualitative and quantitative analysis of asparagine and citrulline using the Platinum EPS reverse-phase C18 column (Alltech Associates. Asparagine alone is eluted earlier than citrulline alone, but when both of them are present in biological samples they may co-elute. HPLC retention times for asparagine and citrulline were influenced by other ureides in the mixture. We found that at various asparagines and citrulline ratios [= 3:1, 1:1, and 1:3; corresponding to 75:25, 50:50, and 25:75 (μMol ml–1/μMol ml–1], the resulting peak exhibited different retention times. Adjustment of ureide ratios as internal standards enables peak identification and quantification. Both chemicals were quantified in xylem sap samples of pecan [Carya illinoinensis (Wangenh. K. Koch] trees. Analysis revealed that tree nickel nutrition status affects relative concentrations of Urea Cycle intermediates, asparagine and citrulline, present in sap. Consequently, we concluded that the HPLC methods are presented to enable qualitative and quantitative analysis of these metabolically important ureides.

  9. Quantitative analysis of mitragynine in human urine by high performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Lu, Shijun; Tran, Buu N; Nelsen, Jamie L; Aldous, Kenneth M

    2009-08-15

    Mitragynine is the primary active alkaloid extracted from the leaves of Mitragyna speciosa Korth, a plant that originates in South-East Asia and is commonly known as kratom in Thailand. Kratom has been used for many centuries for their medicinal and psychoactive qualities, which are comparable to that of opiate-based drugs. Kratom abuse can lead to a detectable content of mitragynine residue in urine. Ultra trace amount of mitragynine in human urine was determined by a high performance liquid chromatography coupled to an electrospray tandem mass spectrometry (HPLC-ESI/MS/MS). Mitragynine was extracted by methyl t-butyl ether (MTBE) and separated on a HILIC column. The ESI/MS/MS was accomplished using a triple quadrupole mass spectrometer in positive ion detection and multiple reactions monitoring (MRM) mode. Ajmalicine, a mitragynine's structure analog was selected as internal standard (IS) for method development. Quality control (QC) performed at three levels 0.1, 1 and 5 ng/ml of mitragynine in urine gave mean recoveries of 90, 109, and 98% with average relative standard deviation of 22, 12 and 16%, respectively. The regression linearity of mitragynine calibration ranged from 0.01 to 5.0 ng/ml was achieved with correlation coefficient greater than 0.995. A detection limit of 0.02 ng/ml and high precision data within-day and between days analysis were obtained.

  10. [Determination of gossypol in edible vegetable oil with high performance liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Zhang, Wenhua; Huang, Chaoqun; Xie, Wen; Shen, Li

    2014-06-01

    A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of gossypol in edible vegetable oil. The sample was extracted with ethyl alcohol by vortex-excited oscillation. The extract was cleaned up by 0.22 microm filter membrane and centrifuged for 5 min at 4 000 r/min after standing in a fridge at 4 degrees C for 30 min. The compound was separated on a C18 column (100 mm x 2.1 mm, 3.5 microm) with acetonitrile and 1% (v/v) formic acid aqueous solution as mobile phase. The detection of gossypol was carried out by LC-MS/MS with positive electrospray ionization under multiple reaction monitoring (MRM) mode using external standard method. The limits of quantification (S/N > 10) of gossypol in edible vegetable oil was 1 mg/kg. The recoveries were from 87.4% to 100% at the spiked levels of 1, 2, 200 mg/kg of gossypol in edible vegetable oil with the relative standard deviations (RSDs) between 3.9% and 12.2%. The method, with high sensitivity, good precision and high recovery, was suitable for the confirmation and quantification of gossypol residue in edible vegetable oil.

  11. Triacylglycerols composition, oxidation and oxidation compounds in camellia oil using liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Zeb, Alam

    2012-07-01

    Camellia seed oil is one of most important edible oil, rich in oleic acid and contains many natural antioxidants with various biological activities. During preparation of foods or storage camellia oil oxidizes by the auto-oxidation and produce oxidized compounds. Traditional analytical techniques like FFA, POV are used for the determination of oxidation and adulteration of oils and fats. These methods were rarely able to detect the oxidized compounds produced and extent of oxidation. This paper presents the uses of liquid chromatography coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS) for the analysis of triacylglycerols (TAGs) composition and evaluation of auto-oxidation and oxidation products of camellia seed oil. The camellia oil was auto-oxidized for 12 months at room temperature. The TAGs were identified from their characteristics fragmentations such as protonated molecular ion, ammonium and sodium adducts, diacylglycerols, epoxy-diacylglycerols fragments and mono-acylglycerol fragments in ESI-MS mass spectra. HPLC-ESI-MS data revealed the separation and identification of 15 TAGs. The major TAGs separated and identified in camellia seed oil were POO, OOO, OLO, PLO/POL, OLL, SOO, ALO and OLLn. The auto-oxidation studies revealed a total loss of LnLLn, LnOLn, LLLn and OLLn amounting about 13.5% total oxidation. The auto-oxidation products were epoxy hydroperoxides, epoxy epidioxides, and mono-epoxides. It was observed that these were characteristic compounds produced in high oleic oils. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  12. A validated new method for nevirapine quantitation in human plasma via high-performance liquid chromatography.

    Science.gov (United States)

    Silverthorn, Courtney F; Parsons, Teresa L

    2006-01-01

    A fully validated and clinically relevant assay was developed for the assessment of nevirapine concentrations in neonate blood plasma samples. Solid-phase extraction with an acid-base wash series was used to prepare subject samples for analysis. Samples were separated by high performance liquid chromatography and detected at 280 nm on a C8 reverse-phase column in an isocratic mobile phase. The retention times of nevirapine and its internal standard were 5.0 and 6.9 min, respectively. The method was validated by assessment of accuracy and precision (statistical values 0.996) and the average recovery was 93% (n = 18). The lower limit of quantification (relative standard deviation <20%) was determined to be 25 ng/mL for 50 microL of plasma, allowing detection of as little as 1.25 ng of nevirapine in a sample. This value represents an increase in sensitivity of up to 30-fold over previously published methods. Copyright 2005 John Wiley & Sons, Ltd.

  13. Effect of first dimension phase selectivity in online comprehensive two dimensional liquid chromatography (LC × LC)

    Science.gov (United States)

    Gu, Haiwei; Huang, Yuan; Filgueira, Marcelo; Carr, Peter W.

    2012-01-01

    In this study, we examined the effect of first dimension column selectivity in reversed phase (RP) online comprehensive two dimensional liquid chromatography (LC × LC). The second dimension was always a carbon clad metal oxide reversed phase material. The hydrophobic subtraction model (HSM) and the related phase selective triangles were used to guide the selection of six different RP first dimension columns. Various kinds of samples were investigated and thus two different elution conditions were needed to cause full elution from the first dimension columns. We compared LC × LC chromatograms, contours plots, and fcoverage plots by measuring peak capacities, peak numbers, relative spatial coverage, correlation values, etc. The major finding of this study is that the carbon phase due to its rather different selectivity from other reversed phases is reasonably orthogonal to a variety of common types of bonded reversed phases. Thus quite surprisingly the six different first dimension stationary phases all showed generally similar separation patterns when paired to the second dimension carbon phase. This result greatly simplifies the task of choosing the correct pair of phases for RP × RP. PMID:21840009

  14. Analysis of Fluconazole in Human Urine Sample by High Performance Liquid Chromatography Method

    Science.gov (United States)

    Hermawan, D.; Ali, N. A. Md; Ibrahim, W. A. Wan; Sanagi, M. M.

    2013-04-01

    A method for determination of fluconazole, antifungal drug in human urine by using reversed-phased high performance liquid chromatography (RP-HPLC) with ultraviolet (UV) detector was developed. Optimization HPLC conditions were carried out by changing the flow rate and composition of mobile phase. The optimum separation conditions at a flow rate 0.85 mL/min with a composition of mobile phase containing methanol:water (70:30, v/v) with UV detection at a wavelength 254 nm was able to analyze fluconazole within 3 min. The excellent linearity was obtained in the range of concentration 1 to 10 μg/mL with r2 = 0.998. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.39 μg/mL and 1.28 μg/mL, respectively. Solid phase extraction (SPE) method using octadecylsilane (C18) as a sorbent was used to clean-up and pre-concentrated of the urine sample prior to HPLC analysis. The average recoveries of fluconazole in spiked urine sample was 72.4% with RSD of 3.21% (n=3).

  15. Determination of isotopically labelled monoesterphthalates in urine by high performance liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Anderson, Warwick A C; Barnes, Karen A; Castle, Laurence; Damant, Andrew P; Scotter, Michael J

    2002-09-01

    A method of analysis for monoesters of phthalic acid ('monoesterphthalates') in human urine has been developed. The method was needed to determine the hydrolysis and excretion efficiency of isotopically-labelled phthalate diesters ('phthalates') when they were fed to volunteers as part of a biomarker study to estimate total exposure to phthalates. The targeted substances were 13C-monobutylphthalate (MBP), 2H4-monobutylphthalate (MBP), 2H4-monobenzylphthalate (MBeP), 13C-monocyclohexylphthalate (MCHP), 13C-monoethylhexylphthalate (MEHP), and 13C-monoisodecylphthalate (MIDP). The monoesters in urine were deconjugated enzymatically, extracted into solvent, and then determined by high performance liquid chromatography-mass spectrometry (LC-MS) using atmospheric pressure chemical ionisation in the negative ion mode. The limits of determination were 10 ng ml(-1) for MBP, MCHP, MBeP and MEHP, and 40 ng ml(-1) for MIDP. The recovery from urine spiked at 100 ng ml(-1) was in the range from 70 to 85% except for MIDP which was lower at 55%. The between-batch reproducibility of the analysis was in the range 8 to 17% (n = 6 batches on separate days).

  16. Analysis of plant nucleotide sugars by hydrophilic interaction liquid chromatography and tandem mass spectrometry.

    Science.gov (United States)

    Ito, Jun; Herter, Thomas; Baidoo, Edward E K; Lao, Jeemeng; Vega-Sánchez, Miguel E; Michelle Smith-Moritz, A; Adams, Paul D; Keasling, Jay D; Usadel, Björn; Petzold, Christopher J; Heazlewood, Joshua L

    2014-03-01

    Understanding the intricate metabolic processes involved in plant cell wall biosynthesis is limited by difficulties in performing sensitive quantification of many involved compounds. Hydrophilic interaction liquid chromatography is a useful technique for the analysis of hydrophilic metabolites from complex biological extracts and forms the basis of this method to quantify plant cell wall precursors. A zwitterionic silica-based stationary phase has been used to separate hydrophilic nucleotide sugars involved in cell wall biosynthesis from milligram amounts of leaf tissue. A tandem mass spectrometry operating in selected reaction monitoring mode was used to quantify nucleotide sugars. This method was highly repeatable and quantified 12 nucleotide sugars at low femtomole quantities, with linear responses up to four orders of magnitude to several 100pmol. The method was also successfully applied to the analysis of purified leaf extracts from two model plant species with variations in their cell wall sugar compositions and indicated significant differences in the levels of 6 out of 12 nucleotide sugars. The plant nucleotide sugar extraction procedure was demonstrated to have good recovery rates with minimal matrix effects. The approach results in a significant improvement in sensitivity when applied to plant samples over currently employed techniques. Copyright © 2013 Elsevier Inc. All rights reserved.

  17. Simultaneous analysis of multiple neurotransmitters by hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry.

    Science.gov (United States)

    Tufi, Sara; Lamoree, Marja; de Boer, Jacob; Leonards, Pim

    2015-05-22

    Neurotransmitters are endogenous metabolites that allow the signal transmission across neuronal synapses. Their biological role is crucial for many physiological functions and their levels can be changed by several diseases. Because of their high polarity, hydrophilic interaction liquid chromatography (HILIC) is a promising tool for neurotransmitter analysis. Due to the large number of HILIC stationary phases available, an evaluation of the column performances and retention behaviors has been performed on five different commercial HILIC packing materials (silica, amino, amide and two zwitterionic stationary phases). Several parameters like the linear correlation between retention and the distribution coefficient (logD), the separation factor k and the column resolution Rs have been investigated and the column performances have been visualized with a heat map and hierarchical clustering analysis. An optimized and validated HILIC-MS/MS method based on the ZIC-cHILIC column is proposed for the simultaneous detection and quantification of twenty compounds consisting of neurotransmitters, precursors and metabolites: 3-methoxytyramine (3-MT), 5-hydroxyindoleacetic acid (5-HIAA), 5-hydroxy-L-tripthophan, acetylcholine, choline, L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine, epinephrine, γ-aminobutyric acid (GABA), glutamate, glutamine, histamine, histidine, L-tryptophan, L-tyrosine, norepinephrine, normetanephrine, phenylalanine, serotonin and tyramine. The method was applied to neuronal metabolite profiling of the central nervous system of the freshwater snail Lymnaea stagnalis. This method is suitable to explore neuronal metabolism and its alteration in different biological matrices. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Detection of HEMA in self-etching adhesive systems with high performance liquid chromatography

    Science.gov (United States)

    Panduric, V.; Tarle, Z.; Hameršak, Z.; Stipetić, I.; Matosevic, D.; Negovetić-Mandić, V.; Prskalo, K.

    2009-04-01

    One of the factors that can decrease hydrolytic stability of self-etching adhesive systems (SEAS) is 2-hydroxymethylmethacrylate (HEMA). Due to hydrolytic instability of acidic methacrylate monomers in SEAS, HEMA can be present even if the manufacturer did not include it in original composition. The aim of the study was to determine the presence of HEMA because of decomposition by hydrolysis of methacrylates during storage, resulting with loss of adhesion strength to hard dental tissues of the tooth crown. Three most commonly used SEAS were tested: AdheSE ONE, G-Bond and iBond under different storage conditions. High performance liquid chromatography analysis was performed on a Nucleosil C 18-100 5 μm (250 × 4.6 mm) column, Knauer K-501 pumps and Wellchrom DAD K-2700 detector at 215 nm. Data were collected and processed by EuroCrom 2000 HPLC software. Calibration curves were made related eluted peak area to known concentrations of HEMA (purchased from Fluka). The elution time for HEMA is 12.25 min at flow rate 1.0 ml/min. Obtained results indicate that no HEMA was present in AdheSE ONE because methacrylates are substituted with methacrylamides that seem to be more stable under acidic aqueous conditions. In all other adhesive systems HEMA was detected.

  19. [Determination of 25 quinolones in cosmetics by liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Lin, Li; Zhang, Yi; Tu, Xiaoke; Xie, Liqi; Yue, Zhenfeng; Kang, Haining; Wu, Weidong; Luo, Yao

    2015-03-01

    An analytical method was developed for the simultaneous determination of 25 quinolones, including danofloxacin mesylate, enrofloxacin, flumequine, oxloinic acid, ciprofloxacin, sarafloxacin, nalidixic acid, norfloxacin, and ofloxacin etc in cosmetics using direct extraction and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). Cosmetic sample was extracted by acidified acetonitrile, defatted by n-hexane and separated on Poroshell EC-C18 column with gradient elution program using acetonitrile and water (both containing 0. 1% formic acid) as the mobile phases and analyzed by LC-ESI-MS/MS under the positive mode using multiple reaction monitoring (MRM). The interference of matrix was reduced by the matrix-matched calibration standard curve. The method showed good linearities over the range of 1-200 mg/kg for the 25 quinolones with good linear correlation coefficients (r ≥ 0.999). The method detection limit of the 25 quinolones was 1.0 mg/kg, and the recoveries of all analytes in lotion, milky and cream cosmetics matrices ranged from 87.4% to 105% at the spiked levels of 1, 5 and 10 mg/kg with the relative standard deviations (RSD) of 4.54%-19.7% (n = 6). The results indicated that this method is simple, fast and credible, and suitable for the simultaneous determination of the quinolones in the above three types of cosmetics.

  20. Simultaneous determination of 11 preservatives in cosmetics by high-performance liquid chromatography.

    Science.gov (United States)

    Aoyama, Airin; Doi, Takahiro; Tagami, Takaomi; Kajimura, Keiji

    2014-10-01

    Preservatives prevent the growth of microorganisms in foods, pharmaceuticals and cosmetics. There exist numerous restrictions regarding the maximum allowable levels of preservatives in cosmetics. We analyzed 11 regulated preservatives in commercial cosmetics and manufacturers need to analyze their products for quality control purposes. However, methods used in previous studies to date have been inadequate for use by public institutions and manufacturers. Therefore, an effective, scalable method for the analysis of preservatives in cosmetics is required. We developed a novel method for the simultaneous determination of 11 regulated preservatives in cosmetics by high-performance liquid chromatography (HPLC). We applied the samples to a C18 column in a simple mobile phase (5 mmol/L ammonium formate solution and acetonitrile) with gradient elution at a flow rate of 1.0 mL/min at a single wavelength (230 nm). The correlation coefficients of the calibration curves were >0.997. The percent recoveries were 92.8-111.9% and the relative standard deviations were 1.9. Because of the simple conditions for isolation and complete separation, the HPLC method can be effectively applied to the analysis of preservatives in commercially retailed cosmetics. © The Author [2013]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  1. Quantitative Determination of Compounds from Akebia quinata by High-Performance Liquid Chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Yen, Nguyen; Thu, Nguyen; Zhao, Bing Tian; Woo, Mi Hee; Min, Byung Sun [Catholic Univ. of Daegu, Gyeongsan (Korea, Republic of); Lee, Jae Hyun [Dongguk Univ., Yongin (Korea, Republic of); Kim, Jeong Ah [Kyungpook National Univ., Daegu (Korea, Republic of); Son, Jong Keun [Yeungnam Univ., Gyeongsan (Korea, Republic of); Choi, Jae Sui [Pukyung National Univ., Busan (Korea, Republic of); Woo, Eun Rhan [Chosun Univ., Gwangju (Korea, Republic of)

    2014-07-15

    To provide the scientific corroboration of the traditional uses of Akebia quinata (Thunb.) Decne., a detailed analytical examination of A. quinata stems was carried out using a reversed-phase high performance liquid chromatography (RP-HPLC) method coupled to photodiode array detector (PDA) for the simultaneous determination of four phenolic substances; cuneataside D, 2-(3,4-dihydroxyphenyl)ethyl-O-β-D-glucopyranoside, 3-caffeoylquinic acid and calceolarioside B. Particular attention was focused on the main compound, 3-caffeoylquinic acid, which has a range of biological functions. In addition, 2-(3,4-dihydroxyphenyl)ethyl-O-β-D-glucopyranoside was considered as a discernible marker of A. quinata from its easy confuse plants. The contents of compounds 2 and 3 ranged from 0.72 to 2.68 mg/g and from 1.66 to 5.64 mg/g, respectively. The validation data indicated that this HPLC/PDA assay was used successfully to quantify the four phenolic compounds in A. quinata from different locations using relatively simple conditions and procedures. The pattern-recognition analysis data from 53 samples classified them into two groups, allowing discrimination between A. quinata and comparable herbs. The results suggest that the established HPLC/PDA method is suitable for quantitation and pattern-recognition analyses for a quality evaluation of this medicinal herb.

  2. Determination of 5-methylcytosine from plant DNA by high-performance liquid chromatography.

    Science.gov (United States)

    Wagner, I; Capesius, I

    1981-06-26

    The relative amounts of the five nucleosides (deoxycytidine, 5-methyldeoxycytidine, deoxyadenosine, deoxyguanosine and thymidine) in the DNA of nine plant species, one plant satellite DNA, and one animal species were determined by high performance liquid chromatography. The method allows the clean separation of the nucleosides from 10 microgram samples with 15 min. The following values for the proportion of methylated cytosines among all cytosines were obtained: Lobularia maritima 18.5%, Nicotiana tabacum 32.6%, Pisum sativum 23.2%, Rhinanthus minor 29.2%, Sinapsis alba 12.2%, Vicia faba 30.5%, Viscum album 23.2%, Cymbidium pumilum 18.8%, Cymbidium pumilum AT-rich satellite DNA 15.8%, Triticum aestivum 22.4%. DNA of an animal, the gerbil, Meriones unguiculatus, had a methylation percentage of 3.1%. An estimate of the GC content based on the buoyant density of DNA tends to be lower than the actual value, an estimate based on the melting temperature tends to be higher. This supports the finding by other authors that DNA methylation decreases the buoyant density and may increase the melting temperature at high m5C concentration.

  3. Simultaneous detection of water-soluble vitamins using the High Performance Liquid Chromatography (HPLC - a review

    Directory of Open Access Journals (Sweden)

    Rosemond Godbless Dadzie

    2014-01-01

    Full Text Available The water-soluble vitamins (WSV: ascorbic acid (vitamin C, thiamine (B1, riboflavin (B2, niacin (B3, panthothenic acid (B5, pyridoxine, and pyridoxal (B6, folic acid (B9, biotin(B8 , and B12 are very essential in the diet of humankind. As a result of ever increasing pressures from both consumers and legal enforcers, to specify accurately nutritive compositions of WSV that are present in food materials, many researchers have attempted to fill this niche through the provision of highly sensitive and rapid high performance liquid chromatography (HPLC procedures. In view of the health benefits of WSV, a replete of HPLC methods have been developed for simultaneous determination of their contents in nature and fortified food samples, nutritional supplements, as well as blood plasmas. The rate of losses of these vitamins during food processing and analysis, in addition to their transient dynamics, presents complexities in developing a highly sensitive HPLC procedure for their simultaneous separations and assays. This review critically assesses the different HPLC procedures developed by researchers and available in the open literature for simultaneous determination of water-soluble vitamins (WSV in dried tropical fruits materials. The study revealed that not a single chromatographic run developed by researchers can simultaneously elute all the WSV at a time. However, the HPLC procedures that are capable of determining all the WSV were coupled with electrospray ionization mass spectroscopy (ESI-MS, thus making the set-up expensive.

  4. Analysis of serotonin concentrations in human milk by high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Chiba, Takeshi; Maeda, Tomoji; Tairabune, Tomohiko; Tomita, Takashi; Sanbe, Atsushi; Takeda, Rika; Kikuchi, Akihiko; Kudo, Kenzo

    2017-03-25

    Serotonin (5-hydroxytryptamine, 5-HT) plays an important role in milk volume homeostasis in the mammary gland during lactation; 5-HT in milk may also affect infant development. However, there are few reports on 5-HT concentrations in human breast milk. To address this issue, we developed a simple method based on high-performance liquid chromatography with fluorescence detection (HPLC-FD) for measuring 5-HT concentrations in human breast milk. Breast milk samples were provided by four healthy Japanese women. Calibration curves for 5-HT in each sample were prepared with the standard addition method between 5 and 1000 ng/ml, and all had correlation coefficients >0.999. The recovery of 5-HT was 96.1%-101.0%, with a coefficient of variation of 3.39%-8.62%. The range of 5-HT concentrations estimated from the calibration curves was 11.1-51.1 ng/ml. Thus, the HPLC-FD method described here can effectively extract 5-HT from human breast milk with high reproducibility. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Rapid Analysis of Cefazolin in Serum by High-Pressure Liquid Chromatography

    Science.gov (United States)

    Wold, John S.

    1977-01-01

    A high-pressure liquid chromatography (HPLC) method has been developed for the analysis of cefazolin in serum. Serum was deproteinized by the addition of 6% trichloroacetic acid and injected onto a reverse-phase column with a mobile phase of 10 to 15% methanol in 1% aqueous acetic acid. Cefazolin chromatographed without interference from ultraviolet-absorbing components of serum, with a retention time of 3.1 min. Standard curves comparing peak area with concentration prepared from dog or human sera were linear over a range of 1.6 to 200 μg/ml. Results from the HPLC assay were compared with microbiological assays (cylinder plate method) on both standard serum samples and sera from dogs and human subjects receiving intramuscular cefazolin. The HPLC method was somewhat more accurate in comparison with the microbiological assay performed on serum samples of known concentration. The comparison of results from an analysis of serum levels of dogs or human subjects receiving cefazolin indicated that the two methods would lead to identical conclusions concerning pharmacokinetics or the achievement of therapeutic serum levels. The HPLC assay method presents an alternative to conventional microbiological assays, with marked improvement in speed (30 min) and considerable potential for future development. PMID:836007

  6. Determination of miloxacin and metabolites in human serum and urine by high-pressure liquid chromatography.

    Science.gov (United States)

    Yoshitake, A; Kawahara, K; Shono, F; Umeda, I; Izawa, A; Komatsu, T

    1980-01-01

    A sensitive and reliable high-pressure liquid chromatography (HPLC) assay for miloxacin and its two principal metabolites, 5,8-dihydro-8-oxo-2H-1,3-dioxolo[4,5-g]quinoline-7-carboxylic acid (M-1) and 1,4-dihydro-1,6-dimethoxy-7-hydroxy-4-oxoquinoline-3-carboxylic acid (M-2), in human serum and urine was developed. A strong anion-exchange Zipax SAX column using a mobile phase of 0.01 M citric acid solution containing 0.03 M sodium nitrate with pH 5.0 was used to achieve separation of the three compounds. The retention times of miloxacin, M-1, and M-2 were 3.8, 9.3, and 5.9 min, respectively. Serum and urine concentrations of these compounds as low as 10 ng/ml were measured. When results from the HPLC assay were compared with those from the microbiological assay of serum and urine samples from human subjects receiving miloxacin orally, the correlation coefficients were 0.94 for the serum and 0.99 for the urine. The HPLC assay method presents an alternative to the microbiological assay and permits future pharmacokinetic investigations of miloxacin. PMID:7416751

  7. COMPARISON OF CAROTENOID CONTENT IN TOMATO, TOMATO PULP AND KETCHUP BY LIQUID CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    J. J. T. GAMA

    2009-01-01

    Full Text Available

    Although tomatoes are commonly consumed fresh, over 80 % the consumption of tomatoes is in the form of processed products such as tomato pulp, ketchup, juice and sauce. Research has indicated the potential health benefits of a diet rich in tomatoes and tomato products. The present study was carried out to determine the carotenoid content of fresh tomato, tomato pulp and ketchup by high performance liquid chromatography. The major differences among these products were in the concentration of some of the pigments. Tomato had all-trans-lycopene (1046-1099 μg/g DW, cislycopene (125-132 μg/g DW and all-trans- -carotene (45-59 μg/g DW as principal carotenoids. Tomato pulp and ketchup had all-trans-lycopene (951-999 μg/g DW and 455-476 μg/g DW, all-trans- -carotene (76-88 DW μg/g and 20-27 DW μg/g and cis-lycopene (71-83 μg/g DW and 14-25 μg/g DW as the main pigments, respectively. They also contained other carotenoids in much smaller amounts (lycoxanthin, zeaxanthin, anteraxanthin, lutein, -carotene, -carotene and phytofluene.

  8. Simultaneous determination of eleven characteristic lignans in Schisandra chinensis by high-performance liquid chromatography.

    Science.gov (United States)

    Hu, Junyang; Mao, Chunqin; Gong, Xiaodong; Lu, Tulin; Chen, Han; Huang, Zhijun; Cai, Baochang

    2013-04-01

    Schisandra chinensis, one of the well-known traditional Chinese herbal medicines, is derived from the dry ripe fruits of Schisandra chinensis (Turcz.) Baill. according to the 9th China Pharmacopeia. Lignans are the main components isolated from extracts of S. chinensis and their content varies depending on where S. chinensis was collected. We have established a qualitative and quantitative method based on the bioactive lignans for control of the quality of S. chinensis from different sources. To develop a high-performance liquid chromatography method, an Elite ODS C18 column (250 mm Χ 4.6 mm, 5μm) at a column temperature of 30°C and flow rate of 1.0ml/min using acetonitrile (A) and water (B) as the mobile phase with a linear gradient and the peaks were monitored at 217 nm. All calibration curves showed good linearity (r ≥ 0.9995) within test ranges. This method showed good repeatability for the quantification of these eleven components in S. chinensis with intra- and inter-day relative standard deviations less than 0.43% and 1.21%, respectively. In the recovery test, results of accuracy ranged from 99.51% to 101.31% with RSD values less than 2. The validated method can be successfully applied to quantify the eleven investigated components in 22 samples of S. chinensis from different sources.

  9. Determination of lipid oxidation level in broiler meat by liquid chromatography.

    Science.gov (United States)

    Marcincák, Slavomir; Sokol, Jozef; Bystrický, Pavel; Popelka, Peter; Turek, Peter; Bhide, Mangesh; Máté, Dionýz

    2004-01-01

    An assay was conducted for the determination of malondialdehyde (MDA) levels in broiler meat. The method involves extraction of tissues with trichloroacetic acid (TCA) and reaction of the TCA extract with 2,4-dinitrophenylhydrazine (DNPH). After separation of the MDA-DNPH complex using a solid-phase extraction C18 column, samples were eluted with 1 mL acetonitrile. Aliquots of 20 microL acetonitrile were analyzed by liquid chromatography on reversed-phase C18 column (3 microm) with UV detection. The products were eluted isocratically with the mobile phase containing acetonitrile-water-acetic acid (39 + 61 + 0.2, v/v/v). The retention time for MDA-DNPH was 6.5 min, and the detection limit was 3.5 microg/kg. Two extraction methods (cold and hot) were also used in the study. The results showed that hot extraction increased results about 55.8% and recovery from samples spiked with 116.6 microg/kg was lower (74.6%) in comparison with cold extraction (94.7%).

  10. Determination of vinblastine in tumour tissue with liquid chromatography-high resolution mass spectrometry.

    Science.gov (United States)

    Kosjek, Tina; Dolinšek, Tanja; Gramec, Darja; Heath, Ester; Strojan, Primož; Serša, Gregor; Čemažar, Maja

    2013-11-15

    There are virtually no analytical methods that describe determination of vinblastine and other vinca alkaloids in tumour tissue, albeit quantitative data on tumour drug amount is essential for maximal benefit of a particular anticancer treatment. The analytical method presented herein uses state-of-the-art sample preparation, separation and detection techniques to allow sensitive and selective determination of vinblastine in tumour tissue. After cryogenic grinding and sonication, tumour suspensions were extracted by Oasis MAX solid phase extraction and analysed for vinblastine with ultra-high performance liquid chromatography coupled to positive electrospray ionisation-high resolution mass spectrometric detection. The developed analytical method quantifies vinblastine down to 23 ng/g tumour tissue and shows satisfactory linearity (r(2)>0.99), precision (1.1-8.2%), accuracy (98%) and high selectivity with almost complete absence of matrix effects. The proposed method was found suitable to follow vinblastine levels in mice tumours and could be used to support preclinical pharmacologic studies. © 2013 Elsevier B.V. All rights reserved.

  11. High perfomance liquid chromatography fingerprint analysis for quality control of brotowali (Tinospora crispa)

    Science.gov (United States)

    Syarifah, V. B.; Rafi, M.; Wahyuni, W. T.

    2017-05-01

    Brotowali (Tinospora crispa) is widely used in Indonesia as ingredient of herbal medicine formulation. To ensure the quality, safety, and efficacy of herbal medicine products, its chemical constituents should be continuously evaluated. High performance liquid chromatography (HPLC) fingerprint is one of powerful technique for this quality control process. In this study, HPLC fingerprint analysis method was developed for quality control of brotowali. HPLC analysis was performed in C18 column and detection was performed using photodiode array detector. The optimum mobile phase for brotowali fingerprint was acetonitrile (ACN) and 0.1% formic acid in gradient elution mode at a flow rate of 1 mL/min. The number of peaks detected in HPLC fingerprint of brotowali was 32 peaks and 23 peaks for stems and leaves, respectively. Berberine as marker compound was detected at retention time of 20.525 minutes. Evaluation of analytical performance including precision, reproducibility, and stability prove that this HPLC fingerprint analysis was reliable and could be applied for quality control of brotowali.

  12. Methacrylate-bonded covalent-organic framework monolithic columns for high performance liquid chromatography.

    Science.gov (United States)

    Liu, Li-Hua; Yang, Cheng-Xiong; Yan, Xiu-Ping

    2017-01-06

    Covalent-organic frameworks (COFs) are a newfangled class of intriguing microporous materials. Considering their unique properties, COFs should be promising as packing materials for high performance liquid chromatography (HPLC). However, the irregular shape and sub-micrometer size of COFs synthesized via the traditional methods render the main obstacles for the application of COFs in HPLC. Herein, we report the preparation of methacrylate-bonded COF monolithic columns for HPLC to overcome the above obstacles. The prepared COF bonded monolithic columns not only show good homogeneity and permeability, but also give high column efficiency, good resolution and precision for HPLC separation of small molecules including polycyclic aromatic hydrocarbons, phenols, anilines, nonsteroidal anti-inflammatory drugs and benzothiophenes. Compared with the bare polymer monolithic column, the COF bonded monolithic columns show enhanced hydrophobic, π-π and hydrogen bond interactions in reverse phase HPLC. The results reveal the great potential of COF bonded monoliths for HPLC and COFs in separation sciences. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. [Determination of ribavirin and amantadine in chicken by ultra performance liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Yun, Huan; Cui, Fengyun; Yan, Hua; Liu, Xin; He, Yue; Zhang, Zhaohui

    2013-08-01

    A method for the determination of ribavirin and amantadine in chicken has been developed by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/ MS). After extracted by 1% (volume percentage) trichloroacetic acid solution-acetonitrile (1:1, v/v) and purified by a Supelco LC-SCX cartridge, the samples were loaded onto an Acquity UPLC BEH Hillic column (150 mm x 2.1 mm, 1.7 microm) and separated with gradient elution. The electrospray was operated in the positive mode and the samples were monitored by the multiple reaction monitoring (MRM) mode. The limits of quantification (LOQs, S/N = 10) of ribavirin and amantadine were 4.0 microg/kg. The calibration curves showed good linearity in the range of 10.0 - 100.0 microg/L, and the correlation coefficients (r(2)) were not lower than 0.99. When the spiked levels were 4.0, 8.0 and 20.0 microg/kg, the recoveries of ribavirin and amantadine in chicken ranged from 78% to 102.5%, with the relative standard deviations (RSDs) of 2.2% - 7.6%. The results indicate that the method is simple, rapid, sensitive and suitable for the qualitative and quantitative analyses of ribavirin and amantadine in chicken samples.

  14. Quantitation of retinaldehyde in small biological samples using ultrahigh-performance liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Wang, Jinshan; Yoo, Hong Sik; Obrochta, Kristin M; Huang, Priscilla; Napoli, Joseph L

    2015-09-01

    We report an ultrahigh-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method to quantify all-trans-retinal in biological samples of limited size (15-35mg), which is especially advantageous for use with adipose. To facilitate recovery, retinal and the internal standard 3,4-didehydroretinal were derivatized in situ into their O-ethyloximes. UHPLC resolution combined with high sensitivity and specificity of MS/MS allowed quantification of retinal-O-ethyloximes with a 5-fmol lower limit of detection and a linear range from 5fmol to 1pmol. This assay revealed that extraocular concentrations of retinal range from approximately 2 to 40pmol/g in multiple tissues-the same range as all-trans-retinoic acid. All-trans-retinoic acid has high affinity (kd⩽0.4nM) for its nuclear receptors (RARα, -β, and -γ), whereas retinal has low (if any) affinity for these receptors, making it unlikely that these retinal concentrations would activate RAR. We also show that the copious amount of vitamin A used in chow diets increases retinal in adipose depots 2- to 5-fold relative to levels in adipose of mice fed a vitamin A-sufficient diet, as recommended for laboratory rodents. This assay also is proficient for quantifying conversion of retinol into retinal in vitro and, therefore, provides an efficient method to study metabolism of retinol in vivo and in vitro. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. Simultaneous characterization and quantification of 17 main compounds in Rabdosia rubescens by high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Sen Guo

    2017-04-01

    Full Text Available Rabdosia rubescens is a healthy herbal tea and well-known Chinese medicinal herb. To evaluate the quality of R. rubescens from China, a high performance liquid chromatography method with dual-wavelength detection was developed and validated. The method was successfully applied for the simultaneous characterization and quantification of 17 main constituents from four different cultivation regions in China. Under optimal conditions, analysis was performed on a Luna C-18 column and gradient elution with a solvent system of acetonitrile and 0.5% (v/v acetic acid–water at a flow rate of 1.0 mL/min and wavelength of 220 nm and 280 nm. All standard calibration curves exhibited good linearity (r2 > 0.9992 within the test ranges. The precision was evaluated by intraday and interday tests, which revealed relative standard deviation values within the ranges of 0.57–2.35% and 0.52–3.40%, respectively. The recoveries were in the range of 96.37–101.66%. The relative standard deviation values for stability and repeatability were < 5%. The contents of some compounds were low and varied with different cultivars. The proposed method could serve as a prerequisite for quality control of R. rubescens materials and products.

  16. Quantitative bioanalysis of peptides by liquid chromatography coupled to (tandem) mass spectrometry.

    Science.gov (United States)

    van den Broek, Irene; Sparidans, Rolf W; Schellens, Jan H M; Beijnen, Jos H

    2008-09-01

    With the growing interest for peptides and proteins in different kinds of fields, e.g. pharmacy, clinical diagnostics or food industry, the quantification of these compounds is becoming more and more important. Quantitative analysis of these analytes in biological matrices, however, remains a challenging task, due to the complexity of both the matrix and the analytical characteristics of these large bio-molecules. Liquid chromatography coupled to (tandem) mass spectrometry (LC-MS or LC-MS/MS) is the preferred analytical technique for peptide analysis as it allows very selective and sensitive measurements. This article summarizes the numerous published LC-MS applications for the quantification of peptides in biological matrices and discusses all different issues herewith concerned. This includes chromatographic aspects as the selection and effects of mobile and stationary phase, flow rate and temperature, as well as mass spectrometric characteristics such as ionization and detection modes, collision-induced dissociation of peptides and factors influencing the mass spectrometric response. For both techniques the main properties of all described methods have been listed, creating a comprehensive overview with the peptide analytes divided into different classes. Likewise, all other issues concerned with quantitative bioanalysis have been evaluated in detail, including extensive consideration of several different applied sample pre-treatment techniques and reflection of subjects as the choice for an internal standard and assay validation. Furthermore, several issues which are of particular interest for the quantitative bioanalysis of peptide compounds like peptide adsorption and degradation have been regarded.

  17. Quantitative analysis of forskolin in Coleus forskohlii (Lamiaceae) by reversed-phase liquid chromatography.

    Science.gov (United States)

    Schaneberg, Brian T; Khan, Ikhlas A

    2003-01-01

    A rapid method was developed for the evaluation of forskolin in Coleus forskohlii Briq. (Lamiaceae). Forskolin was quantitated in the root and stem of dried C. forskohlii and in 17 market products by reversed-phase liquid chromatography (LC) with a photodiode array detector at 210 nm. The temperature was held constant at 30 degrees C, and the retention time of forskolin was approximately 6.8 min. The samples were extracted with acetonitrile by sonication. The precision of the method was confirmed by a standard deviation < 5.0% (n = 3), and forskolin recovery was 99.1%. Limit of detection was 1.5 microg/mL, and the response was linear through zero from 6.3 to 630 microg/mL with a correlation coefficient (R2) of 0.9998. Identity of the marker compound was confirmed by an LC/mass spectrometry experiment. The method was successful in the qualitative and quantitative evaluation of the marker compound in C. forskohlii plant material and in market products claiming to contain C. forskohlii.

  18. Determination of the Thyreostats in Animal Feeding Stuffs Using Liquid Chromatography-Tandem Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Woźniak Barbara

    2014-10-01

    Full Text Available A rapid liquid chromatography tandem mass spectrometry method was developed and validated to detect and confirm five thyreostatic drugs: tapazole, thiouracil, methylthiouracil, propylthiouracil, and phenylthiouracil in animal feeding stuff samples. Thyreostats were extracted from feed with methanol, and then degreasing of the extract with petroleum ether was performed, followed by the derivatisation of the compounds with 3-iodobenzylbromide in basic medium (pH 8.0. The derivatives were extracted with diethyl ether and analysed by gradient elution on a Poroshell 120-EC C18 column with triple quadrupole MS detection with turbo spray source in positive ionisation mode. The method was validated in accordance with the Commission Decision 2002/657/EC. For validation level of 10 ļig kg-1, the recovery ranged from 82% to 97.5% for all examined compounds. The repeatability and reproducibility did not exceed the limit of 20% for all analytes. The linearity was good for all thyreostats in the whole range of tested concentrations, as proved by the correlation coefficients greater than 0.99. The decision limits (CCa ranged from 1.63 ļig kg-1 to 3.95 ļig kg-1, whereas the detection capabilities (CCß ranged from 2.74 ļig kg-1 to 6.73 ļig kg-1. The developed analysis is sensitive and robust, and therefore useful for quantification and confirmation of thyreostats in residue control programme.

  19. Determination of porphyrins in oral bacteria by liquid chromatography electrospray ionization tandem mass spectrometry.

    Science.gov (United States)

    Fyrestam, Jonas; Bjurshammar, Nadja; Paulsson, Elin; Johannsen, Annsofi; Östman, Conny

    2015-09-01

    Biofilms in the oral cavity can be visualized by fluorescence and a common assumption is that the endogenously produced porphyrins in certain bacteria give rise to this fluorescence. Porphyrin content in oral bacteria has been sparingly investigated, and non-selective detection techniques such as utilizing the Soret fluorescence band of porphyrins are often used. In the present study, a quantitative and selective method for the determination of porphyrins in oral bacteria has been developed and validated using high performance liquid chromatography-tandem mass spectrometry. Lysis of bacteria using Tris-EDTA buffer together with ultrasonication showed high microbial killing efficiency ≥99.98%, and sample clean-up using C18-solid phase extraction resulted in low matrix effects ≤14% for all analytes. Using this method, the porphyrin content was determined in the two oral pathogens Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis, as well as for baker's yeast, Saccharomyces cerevisiae. Uroporphyrin, 7-carboxylporphyrin, 6-carboxylporphyrin, coproporphyrin, and protoporphyrin IX were identified in the investigated microorganisms, and it was shown that the porphyrin profile differs between the two bacteria, as well as for S. cerevisiae. To our knowledge, this is the first time the porphyrin profile has been determined for the bacterium A. actinomycetemcomitans.

  20. Quantitative determination of amygdalin epimers from armeniacae semen by liquid chromatography.

    Science.gov (United States)

    Koo, Ja-Yong; Hwang, Eun-Young; Cho, Sonhae; Lee, Je-Hyun; Lee, Yong-Moon; Hong, Seon-Pyo

    2005-01-05

    D-amygdalin and its conversion product, neoamygdalin, were quantitatively analyzed on reverse-phase, high-performance liquid chromatography with an optimized eluent of 10 mM sodium phosphate buffer (pH 3.1) containing 8.5% acetonitrile. Linearity between concentrations and detector responses was obtained in the range from 0.05 to 0.5 mM. The detection limits for D-amygdalin and neoamygdalin were approximately 5 microM per injected amount. Armeniacae semen contains not only amygdalin but also emlusin, which is an enzyme that hydrolyzes amygdalin. When extracting amygdalin from a whole piece of armeniacae semen in boiling water, there was almost no influence of emulsin; which increased the extraction efficiency. However, conversion of d-amygdalin into neoamygdalin at high temperature was found. In this report, we solved this problem by using 4% citric acid as an extractant. This solution also prevented the extraction process from being affected by emulsin. In addition, the extraction efficiency remained the same as that when methanol was used as an extractant, regardless of the cutting size.