PHYTOCHEMICAL ANALYSIS, Issue 2 2010
Nian Yun Yang
Abstract Introduction , The aerial part Eupatorium lindleyanum is commonly used as an antipyretic and detoxicant clinically in traditional Chinese medicine. Our previous research showed that germacrane sesquiterpene lactones were its main active constituents, so the development of rapid and accurate methods for the identification of the sesquiterpene lactones is of great significance. Objective , To develop an HPLC-PDA-ESI-MS/MS method capable for simple and rapid analysis of germacrane sesquiterpene lactones in the aerial part E. lindleyanum. Methodology , High-performance liquid chromatography-photodiode array detection-electrospray ionization-tandem mass spectrometry was used to analyze germacrane sesquiterpene lactones of Eupatorium lindleyanum. The fragmentation behavior of germacrane sesquiterpene lactones in a Micromass Q/TOF Mass Spectrometer was discussed, and 9 germacrane sesquiterpene lactones were identified by comparison of their characteristic data of HPLC and MS analyses with those obtained from reference compounds. Results , The investigated germacrane sesquiterpene lactones were identified as eupalinolides C (1), 3,-acetoxy-8,-(4,-hydroxy-tigloyloxy)-14-hydroxy-costunolide (2), eupalinolides A (3), eupalinolides B (4), eupalinolides E (5), 3,-acetoxy-8,-(4,-oxo-tigloyloxy)-14-hydroxy-heliangolide (6), 3,-acetoxy-8,-(4,-oxo- tigloyloxy)-14-hydroxy-costunolide (7), hiyodorilactone B (8), and 3,-acetoxy-8,-(4,-hydroxy-tigloyloxy)- costunolide (9). Compounds 6, 7 and 9 were reported for the first time. Conclusion , HPLC-PDA-ESI-MS/MS provides a new powerful approach to identify germacrane sesquiterpene lactones in E. lindleyanum rapidly and accurately. Copyright © 2009 John Wiley & Sons, Ltd. [source]

An HPLC-MS method for simultaneous estimation of ,,, -arteether and its metabolite dihydroartemisinin, in rat plasma for application to pharmacokinetic study

BIOMEDICAL CHROMATOGRAPHY, Issue 7 2003
M. Rajanikanth
Abstract This manuscript reports, the development and validation of a sensitive and selective assay method for simultaneous determination of ,,, -arteether and its metabolite dihydroartemisinin (DHA) in rat plasma by liquid chromatography,mass spectrometry. Chromatographic separations were achieved by gradient elution of the analytes with an initial composition of methanol,potassium acetate buffer (pH 4; 73:27, v/v) to 100% methanol in 3 min and maintained for 5 min on a Spheri-10, RP18 (100 × 4.6 mm i.d.) column following an RP18 (30 × 4.6 mm i.d.) guard column. The total ef,uent from the column was split so that one-tenth was injected into the electrospray LC/MS interface. ESI-MS analysis was performed using a Micromass Quattro II Triple Quadrupole Mass Spectrometer equipped with an electrospray source. The MS analysis was carried out at cone voltage of 22 V with a scan range of 200,500 Da. The analytes were quanti,ed from the [M+ K]+ ion chromatograms of ,,, -arteether at m/z 352, DHA at m/z 323, artemisinin at m/z 321 and propyl ether analogue of arteether at m/z 365. Liquid,liquid extractions with a combination of n -hexane and hexane,ethyl acetate (8:2) were used to isolate ,,, -arteether and DHA from rat plasma. The method was validated and gave good accuracy and precision for the studied domain. Linearity in serum was observed over the range 4.375,70 ng/mL for a -arteether and 10,160 ng/mL for , -arteether and DHA. Percentage bias (accuracy) and within- and between-assay precision were well within the acceptable range. This method was applied to study the pharmacokinetics following oral administration of ,,, -arteether (30 mg/kg) in rats. Copyright © 2003 John Wiley & Sons, Ltd. [source]

Relation between Developmental Stage, Sensory Properties, and Volatile Content of Organically and Conventionally Grown Pac Choi (Brassica rapa,var.

JOURNAL OF FOOD SCIENCE, Issue 4 2010
Mei Qing Choi)
ABSTRACT:, This study was conducted to identify and quantify the sensory characteristics and chemical profile of organically and conventionally grown pac choi (Brassica rapa,var. Mei Qing Choi), also called bok choy, at 3 stages of growth (2.5, 4.5, and 6.5 wk). Sensory and instrumental data were correlated using partial least squares regression. Pac choi was grown in late spring. Descriptive sensory analysis was conducted by a highly trained panel and compounds were identified and quantified using a gas chromatograph/mass spectrometer. The findings of the study indicate that the differences in sensory characteristics and chemical profiles among stages of growth are more substantial than the differences between organic and conventional production. Green-unripe, musty/earthy, lettuce, and sweet flavors are representative in pac choi at early stages of growth. When older, pac choi has higher intensities of green-grassy/leafy, bitter, cabbage, and sulfur flavors that are associated with the increase of (Z)-3-hexen-1-ol, octyl acetate, 1-nonanol, 2-decanone, 1-penten-3-ol, linalool, camphor, menthol, isobornyl acetate, geranylacetone, and cedrol compounds. Conventional pac choi was higher than organic pac choi in green overall, bitter, and soapy flavors only at 2.5 wk of age. This may be associated with the presence of (Z)-3-hexenal, 2-hexyn-1-ol, and (E)-2-hexenal compounds. Practical Application: The increased popularity of organic production has amplified the need for research that will help in understanding how this production system affects the final quality of food products. This study suggests that the stage of development has a much larger impact on sensory quality than organic or conventional growing of pac choi. Findings from this study promote consumer choice by showing that comparable sensory quality can be obtained using either production system, making the ultimate choice not only based on sensory quality but consumer choice related to environmental beliefs or economics. [source]

Volatile metabolite profiling to detect and discriminate stem-end rot and anthracnose diseases of mango fruits

PLANT PATHOLOGY, Issue 6 2006
M. Moalemiyan
The volatile metabolites from the headspace gas of containerised mango (Mangifera indica) cv. Tommy Atkins fruits, surface wounded and inoculated with the two fungal anamorphic pathogens Colletotrichum gloeosporioides and Lasiodiplodia theobromae, or non-inoculated (controls), were profiled using a portable gas chromatograph/mass spectrometer to discriminate diseases of mango. Thirty-four compounds were detected relatively consistently among replicates. Several of these were disease/inoculation-discriminatory and were classified into three groups: (i) compounds unique to only one treatment; (ii) compounds common to two or more treatments, but not to all; and (iii) compounds common to all treatments, but varying in their abundance. Two compounds, 1-pentanol and ethyl boronate, were detected in L. theobromae- inoculated mangoes alone, while thujol was observed only in C. gloeosporioides- inoculated mangoes. Discriminant analysis models based on the abundance of significant mass ions and consistent compounds correctly classified diseases/inoculations in up to 100% of cases. The disease-discriminatory compounds and discriminant analysis models developed here have the potential to be used in the early detection of postharvest diseases of mango fruits after validation under commercial conditions. [source]

Design and implementation of a high-performance CCA event service,

CONCURRENCY AND COMPUTATION: PRACTICE & EXPERIENCE, Issue 9 2009
Ian Gorton
Abstract Event services based on publish,subscribe architectures are well-established components of distributed computing applications. Recently, an event service has been proposed as part of the common component architecture (CCA) for high-performance computing (HPC) applications. In this paper we describe our implementation, experimental evaluation, and initial experience with a high-performance CCA event service that exploits efficient communications mechanisms commonly used on HPC platforms. We describe the CCA event service model and briefly discuss the possible implementation strategies of the model. We then present the design and implementation of the event service using the aggregate remote memory copy interface as an underlying communication layer for this mechanism. Two alternative implementations are presented and evaluated on a Cray XD-1 platform. The performance results demonstrate that event delivery latencies are low and that the event service is able to achieve high-throughput levels. Finally, we describe the use of the event service in an application for high-speed processing of data from a mass spectrometer and conclude by discussing some possible extensions to the event service for other HPC applications. Published in 2009 by John Wiley & Sons, Ltd. [source]

Screening for the calstabin-ryanodine receptor complex stabilizers JTV-519 and S-107 in doping control analysis

DRUG TESTING AND ANALYSIS, Issue 1 2009
Mario Thevis
Abstract Recent studies outlined the influence of exercise on the stability of the skeletal muscle calstabin1-ryanodine receptor1-complex, which represents a major Ca2+ release channel. The progressive modification of the type-1 skeletal muscle ryanodine receptor (RyR1) combined with reduced levels of calstabin1 and phosphodiesterase PDE4D3 resulted in a Ca2+ leak that has been a suggested cause of muscle damage and impaired exercise capacity. The use of 1,4-benzothiazepine derivatives such as the drug candidates JTV-519 and S-107 enhanced rebinding of calstabin1 to RyR1 and resulted in significantly improved skeletal muscle function and exercise performance in rodents. Due to the fact that the mechanism of RyR1 remodelling under exercise conditions were proven to be similar in mice and humans, a comparable effect of JTV-519 and S-107 on trained athletes is expected, making the compounds relevant for doping controls. After synthesis of JTV-519, S-107, and a putative desmethylated metabolite of S-107, target compounds were characterized using nuclear magnetic resonance spectroscopy and electrospray ionization (ESI),high-resolution/high-accuracy Orbitrap mass spectrometry. Collision-induced dissociation pathways were suggested based on the determination of elemental compositions of product ions and H/D-exchange experiments. The most diagnostic product ion of JTV-519 was found at m/z 188 (representing the 4-benzyl-1-methyl piperidine residue), and S-107 as well as its desmethylated analog yielded characteristic fragments at m/z 153 and 138 (accounting for 1-methoxy-4-methylsulfanyl-benzene and 4-methoxy-benzenethiol residues, respectively). The analytes were implemented in existing doping control screening procedures based on liquid chromatography, multiple reaction monitoring and simultaneous precursor ion scanning modes using a triple quadrupole mass spectrometer. Validation items such as specificity, recovery (68,92%), lower limit of detection (0.1,0.2 ng/mL), intraday (5.2,18.5%) and interday (8.7,18.8%) precision as well as ion suppression/enhancement effects were determined. Copyright © 2009 John Wiley & Sons, Ltd. [source]

CEC-ESI ion trap MS of multiple drugs of abuse

ELECTROPHORESIS, Issue 7 2010
Zeineb Aturki
Abstract This article describes a method for the separation and determination of nine drugs of abuse in human urine, including amphetamines, cocaine, codeine, heroin and morphine. This method was based on SPE on a strong cation exchange cartridge followed by CEC-MS. The CEC experiments were performed in fused silica capillaries (100,,m×30,cm) packed with a 3,,m cyano derivatized silica stationary phase. A laboratory-made liquid junction interface was used for CEC-MS coupling. The outlet capillary column was connected with an emitter tip that was positioned in front of the MS orifice. A stable electrospray was produced at nanoliter per minute flow rates applying a hydrostatic pressure (few kPa) to the interface. The coupling of packed CEC columns with mass spectrometer as detector, using a liquid junction interface, provided several advantages such as better sensitivity, low dead volume and independent control of the conditions used for CEC separation and ESI analysis. For this purpose, preliminary experiments were carried out in CEC-UV to optimize the proper mobile phase for CEC analysis. Good separation efficiency was achieved for almost all compounds, using a mixture containing ACN and 25,mM ammonium formate buffer at pH 3 (30:70, v/v), as mobile phase and applying a voltage of 12,kV. ESI ion-trap MS detection was performed in the positive ionization mode. A spray liquid, composed by methanol,water (80:20, v/v) and 1% formic acid, was delivered at a nano-flow rate of ,200,nL/min. Under optimized CEC-ESI-MS conditions, separation of the investigated drugs was performed within 13,min. CEC-MS and CEC-MS2 spectra were obtained by providing the unambiguous confirmation of these drugs in urine samples. Method precision was determined with RSDs values ,3.3% for retention times and ,16.3% for peak areas in both intra-day and day-to-day experiments. LODs were established between 0.78 and 3.12,ng/mL for all compounds. Linearity was satisfactory in the concentration range of interest for all compounds (r2,0.995). The developed CEC-MS method was then applied to the analysis of drugs of abuse in spiked urine samples, obtaining recovery data in the range 80,95%. [source]

CE-LIF-MSn profiling of oligosaccharides in human milk and feces of breast-fed babies

ELECTROPHORESIS, Issue 7 2010
Simone Albrecht
Abstract Mixtures of the complex human milk oligosaccharides (HMOs) are difficult to analyze and gastrointestinal bioconversion products of HMOs may complicate analysis even more. Their analysis, therefore, requires the combination of a sensitive and high-resolution separation technique with a mass identification tool. This study introduces for the first time the hyphenation of CE with an electrospray mass spectrometer, capable to perform multiple MS analysis (ESI-MSn) for the separation and characterization of HMOs in breast milk and feces of breast-fed babies. LIF was used for on- and off-line detections. From the overall 47 peaks detected in off-line CE-LIF electropherograms, 21 peaks could be unambiguously and 11 peaks could be tentatively assigned. The detailed structural characterization of a novel lacto- N -neo-tetraose isomer and a novel lacto- N -fucopentaose isomer was established in baby feces and pointed to gastrointestinal hydrolysis of higher-Mw HMOs. CE-LIF-ESI-MSn presents, therefore, a useful tool which contributes to an advanced understanding on the fate of individual HMOs during their gastrointestinal passage. [source]

A novel methodology for the analysis of membrane and cytosolic sub-proteomes of erythrocytes by 2-DE

ELECTROPHORESIS, Issue 23 2009
Gloria Alvarez-Llamas
Abstract With the aim of studying a wide cohort of erythrocyte samples in a clinical setting, we propose here a novel approach that allows the analysis of both human cytosolic and membrane sub-proteomes. Despite their simple structure, the high content of hemoglobin present in the red blood cells (RBCs) makes their proteome analysis enormously difficult. We investigate here different strategies for isolation of the membrane and cytosolic fractions from erythrocytes and their influence on proteome profiling by 2-DE, paying particular attention to hemoglobin removal. A simple, quick and satisfactory approach for hemoglobin depletion based on HemogloBindÔ reagent was satisfactorily applied to erythrocyte cells, allowing the analysis of the cytosolic sub-proteome by 2-DE without major interference. For membrane proteome, a novel combined strategy based on hypotonic lysis isolation and further purification on minicolumns is described here, allowing detection of high molecular weight proteins (i.e. spectrin, ankyrin) and well-resolved 2-DE patterns. An aliquot of the membrane fraction was also in solution digested and analyzed by nano-LC coupled to an LTQ-Orbitrap mass spectrometer. A total of 188 unique proteins were identified by this approach. This study sets the basis for future clinical studies where the erythrocyte cell may be implicated. [source]

Analyses of gibberellins in coconut (Cocos nucifera L.) water by partial filling-micellar electrokinetic chromatography-mass spectrometry with reversal of electroosmotic flow

ELECTROPHORESIS, Issue 10 2008
Liya Ge
Abstract In this paper, we present the results of simultaneous screening of eight gibberellins (GAs) in coconut (Cocos nucifera L.) water by MEKC directly coupled to ESI-MS detection. During the development of MEKC-MS, partial filling (PF) was used to prevent the micelles from reaching the mass spectrometer as this is detrimental to the MS signal, and a cationic surfactant, cetyltrimethylammonium hydroxide, was added to the electrolyte to reverse the EOF. On the basis of the resolution of the neighboring peaks, different parameters (i.e., the pH and concentration of buffer, surfactant concentrations, length of the injected micellar plug, organic modifier, and applied separation voltage) were optimized to achieve a satisfactory PF-MEKC separation of eight GA standards. Under optimum conditions, a baseline separation of GA standards, including GA1, GA3, GA5, GA6, GA7, GA9, GA12, and GA13, was accomplished within 25,min. Satisfactory results were obtained in terms of precision (RSD of migration time below 0.9%), sensitivity (LODs in the range of 0.8,1.9,,M) and linearity (R2 between 0.981 and 0.997). MS/MS with multiple reaction monitoring (MRM) detection was carried out to obtain sufficient selectivity. PF-MEKC-MS/MS allowed the direct identification and confirmation of the GAs presented in coconut water (CW) sample after SPE, while, the quantitative analysis of GAs was performed by PF-MEKC-MS approach. GA1 and GA3 were successfully detected and quantified in CW. It is anticipated that the current PF-MEKC-MS method can be applicable to analyze GAs in a wide range of biological samples. [source]

Rapid capillary electrophoresis time-of-flight mass spectrometry separations of peptides and proteins using a monoquaternarized piperazine compound (M7C4I) for capillary coatings

ELECTROPHORESIS, Issue 8 2008
Anisa Elhamili
Abstract A monoquaternarized piperazine, 1-(4-iodobutyl) 4-aza-1-azoniabicyclo[2,2,2] octane iodide (M7C4I), has been evaluated as a surface derivatization reagent for CE in combination with TOF MS for the analysis of proteins, peptides, and protein digests. The M7C4I piperazine, at alkaline pH, forms a covalent bond via alkylation of the ionized silanols producing a cationic surface with a highly stable and reversed EOF. The obtained surface yields rapid separations (less than 5,min) of peptides and proteins at acidic pH with high separation efficiencies (up to 1.1×106 plates/m for peptides and up to 1.8×106 plates/m for proteins) and no observed bleeding of the coating reagent into the mass spectrometer. The simplicity of the coating procedure also enables fast (2,min) regeneration of the surface, if necessary. This is useful in the analysis of complex samples in order to prevent possible memory effects. The potential of using M7C4I-coated capillaries for MS analysis of complex samples is demonstrated by the separation of peptides, proteins, and protein digests. Even more, the spectacular thing in which large intact proteins with molecular masses over 0.5,MDa could be separated. The coating showed good ability to handle these large proteins with high efficiency and retained peak shape as demonstrated by separation of IgG1 (150,kDa) and thyroglobulin (669,kDa). [source]

A novel approach for analysis of oligonucleotide,cisplatin interactions by continuous elution gel electrophoresis coupled to isotope dilution inductively coupled plasma mass spectrometry and matrix-assisted laser desorption/ionization mass spectrometry

ELECTROPHORESIS, Issue 7 2008
Wolfram Brüchert
Abstract In this work we present a novel approach for in vitro studies of cisplatin interactions with 8-mer oligonucleotides. The approach is based on the recently developed coupling of continuous elution gel electrophoresis (GE) to an inductively coupled plasma-sector field mass spectrometer (ICP-SFMS) with the aim of monitoring the interaction process between this cytostatic drug and the nucleotides. In contrast to existing methods, the electrophoretic separation conditions used here allow both the determination of the reaction kinetics in more detail as well as the observation of dominant intermediates. Two different nucleotides sequences have been investigated for comparison purposes, one containing two adjacent guanines (5,-TCCGGTCC-3,) and one with a combination of thymine and guanine (5,-TCCTGTCC-3,), respectively. In order to gain further structural information, MALDI-TOF MS measurements have been performed after fraction collection. This allows for identification of the intermediates and the final products and confirms the stepwise coordination of cisplatin via monoadduct to bisadduct formation. Furthermore, the ICP-MS results were quantitatively evaluated in order to calculate the kinetics of the entire process. [source]

Ammonium perfluorooctanoate as a volatile surfactant for the analysis of N -methylcarbamates by MEKC-ESI-MS

ELECTROPHORESIS, Issue 22 2006
Geert Van Biesen
Abstract Ammonium perfluorooctanoate (APFOA) was investigated as an MS-friendly surfactant for the analysis of a mixture of ten N -methylcarbamates with MEKC-ESI-MS. Because of the relatively low boiling point of perfluorooctanoic acid (,190°C), APFOA can be introduced into a mass spectrometer without the adverse effects of less volatile surfactants such as SDS. With a BGE consisting of 50,mM APFOA/isopropanol (IPA) 98:2 and with 30,kV applied, a very fast separation (,6,min) was possible with only one pair of analytes comigrating. Using an experimental design with four factors (voltage, nebulizer pressure, concentration of APFOA, and concentration of IPA) we were able to resolve all analytes in just over 11,min. Sheath liquid composition and flow rate, drying gas temperature and flow rate, and fragmentor voltage were then optimized for maximum signal intensity and S/N. It was found that the faster method gave better S/N because of narrower peak widths, and detection limits in SIM mode were between 0.01 (aldicarb) and 0.08,mg/L (methomyl). Calibration curves were prepared with standards of 0.50, 1.00, and 2.00,mg/L for the analysis of samples obtained after SPE of tap water spiked with the ten N -methylcarbamates at a level of 10,µg/L. All analytes showed very good recoveries (>86%), except for the most polar analyte aldicarb sulfone (recovery of 73%), testifying for the potential use of APFOA for this kind of analyses. [source]

Contribution of Chloroflexus respiration to oxygen cycling in a hypersaline microbial mat from Lake Chiprana, Spain

ENVIRONMENTAL MICROBIOLOGY, Issue 8 2007
Lubos Polerecky
Summary In dense stratified systems such as microbial mats, photosynthesis and respiration are coupled due to a tight spatial overlap between oxygen-producing and -consuming microorganisms. We combined microsensors and a membrane inlet mass spectrometer with two independent light sources emitting in the visible (VIS) and near infrared (NIR) regions to study this coupling in more detail. Using this novel approach, we separately quantified the activity of the major players in the oxygen cycle in a hypersaline microbial mat: gross photosynthesis of cyanobacteria, NIR light-dependent respiration of Chloroflexus -like bacteria (CLB) and respiration of aerobic heterotrophs. Illumination by VIS light induced oxygen production in the top ,1 mm of the mat. In this zone CLB were found responsible for all respiration, while the contribution of the aerobic heterotrophs was negligible. Additional illumination of the mat with saturating NIR light completely switched off CLB respiration, resulting in zero respiration in the photosynthetically active zone. We demonstrate that microsensor-based quantification of gross and net photosyntheses in dense stratified systems should carefully consider the NIR light-dependent behaviour of CLB and other anoxygenic phototrophic groups. [source]

Analysis of endosulfan and its metabolites in rat plasma and selected tissue samples by gas chromatography,mass spectrometry

ENVIRONMENTAL TOXICOLOGY, Issue 1 2005
Melissa P. L. Chan
Abstract A method has been developed for the determination of trace levels of ,-endosulfan, ,-endosulfan, endosulfan sulfate, and endosulfan diol in rat plasma and tissue samples. Endosulfan and its metabolites in the plasma samples were extracted with solid-phase extraction Chromabond-end-capped C18 cartridges and analyzed by a Shimadzu QP-5050A gas chromatograph,mass spectrometer (GCMS) with quadrupole detector in selected-ion-monitoring mode. The analysis of endosulfan and its metabolites in liver and kidney samples involved solvent extraction, Florisil solid-phase-extraction cleanup, and quantitation by GCMS. Recovery experiments for the plasma and tissue samples were conducted over concentration ranges of 10,100 ng mL,1 and 100,1000 ng mL,1, respectively. The method was applied to the analysis of trace levels of endosulfan and its metabolites in plasma and tissue samples collected from an animal study. Trace levels of ,-endosulfan and ,-endosulfan in the ranges of undetectable to 3.11 ,g g,1 and undetectable to 1.19 ,g g,1, respectively, were detected in the kidney samples, whereas trace levels of endosulfan sulfate in the range of 0.02,0.22 ,g g,1 were detected in the liver samples of rats. Neither endosulfan nor its metabolites was detected in any of the plasma samples. © 2005 Wiley Periodicals, Inc. Environ Toxicol 20: 45,52, 2005. [source]

Occurrence and fate of micropollutants in the Vidy Bay of Lake Geneva, Switzerland.

ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 8 2010
Part II: Micropollutant removal between wastewater, raw drinking water
Abstract The occurrence and removal of 58 pharmaceuticals, endocrine disruptors, corrosion inhibitors, biocides, and pesticides, were assessed in the wastewater treatment plant (WWTP) of the city of Lausanne, Switzerland, as well as in the effluent-receiving water body, the Vidy Bay of Lake Geneva. An analytical screening method to simultaneously measure all of the 58 micropollutants was developed based on ultra performance liquid chromatography coupled to a tandem mass spectrometer (UPLC-MS/MS). The selection of pharmaceuticals was primarily based on a prioritization study, which designated them as environmentally relevant for the Lake Geneva region. Except for the endocrine disruptor 17,-ethinylestradiol, all substances were detected in 24-h composite samples of wastewater entering the WWTP or in the treated effluent. Of these compounds, 40% were also detected in raw drinking water, pumped from the lake 3,km downstream of the WWTP. The contributions of dilution and degradation to micropollutant elimination between the WWTP outlet and the raw drinking water intake were established in different model scenarios using hypothetical residence times of the wastewater in Vidy Bay of 1, 4, or 90 d. Concentration decrease due to processes other than dilution was observed for diclofenac, beta-blockers, several antibiotics, corrosion inhibitors, and pesticides. Measured environmental concentrations (MECs) of pharmaceuticals were compared to the predicted environmental concentrations (PECs) determined in the prioritization study and agreed within one order of magnitude, but MECs were typically greater than the corresponding PECs. Predicted no-effect concentrations of the analgesic paracetamol, and the two antibiotics ciprofloxacin and sulfamethoxazole, were exceeded in raw drinking water samples and therefore present a potential risk to the ecosystem. Environ. Toxicol. Chem. 2010; 29:1658,1668. © 2010 SETAC [source]

,, ,-Unsaturated sulfophenylcarboxylates as degradation intermediates of linear alkylbenzenesulfonates: Evidence for ,-oxygenation followed by ,-oxidations by liquid chromatography-mass spectrometry

ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 1 2002
Peter Eichhorn
Abstract Liquid chromatography with an electrospray interface to a mass spectrometer (LC-ES-MS) and LC-ES coupled to a tandem MS (LC-ES-MS/MS) were used to detect and identify intermediates excreted transiently during the aerobic degradation of linear alkylbenzenesulfonates (LAS) in fixed-bed bioreactors (FBBR). The inoculum for the FBBR was the microflora of the River Rhine, Germany. Two major phenomena were observed on the addition of 100 mg/L LAS to the system, sorption and then biodegradation. Disappearance due to sorption was followed in an inhibited FBBR. Biodegradation of LAS started on day 7 and was accompanied by the transient excretion of intermediates, which were later largely degraded. We detected not only the sulfophenylcarboxylates (SPCs) observed previously but also the ,, ,-unsaturated SPCs (SPC-2H), which have not been reported before. Experiments with the (4-sulfophenyl)dodecanes (C12-LAS), which had minor contaminants of C11-LAS, showed C12-, C10-, C8-, C6-, and C4-SPCs when LAS was degraded as well as traces of C9-, C7-, and C5-SPCs. Signals from the SPC-2H species were usually some 10% of those from the corresponding SPCs. Samples from these experiments were also examined by gas chromatography-mass spectrometry (GC-MS), but no desulfonated intermediates were detected. We interpret the data to mean that the only attack on LAS was by ,-oxygenation; there was no visible initial desulfonation. The products of ,-oxygenation were oxidized to the corresponding SPC and subject to ,-oxidation, as evidenced not only by the pattern of C-2 units in the excreted SPCs but also in the corresponding series of SPC-2H, representing the intermediates in ,-oxidation. [source]

Peptides of human gingival crevicular fluid determined by HPLC-ESI-MS

EUROPEAN JOURNAL OF ORAL SCIENCES, Issue 6 2005
Elisabetta Pisano
The acidic-soluble protein content of human gingival crevicular fluid was analyzed by reverse-phase high-performance liquid chromatography (RP-HPLC), and the eluent deriving from the chromatography separation was directly introduced into an ion-trap mass spectrometer through electrospray ionization (ESI-IT MS). By this technique the molecular weight of peptides/proteins was determined with a precision of ,,1/10,000 amu. On the basis of the chromatographic behavior and the knowledge of the molecular mass value, some peptides and proteins soluble in acidic solution were unambiguously recognized. Besides high quantities of human serum albumin, , -defensins 1,4 and minor amounts of cystatin A, statherin, basic PB salivary peptide and other unidentified components were detected. The presence of , -defensins in gingival crevicular fluid is in agreement with their relevant contribution to protein composition deriving from granulocyte secretions. Other peptides and proteins abundant in human saliva, such as proline-rich proteins (PRPs) and histatins, were not detected in gingival crevicular fluid. Further investigations will be necessary to establish the origin of statherin and PB salivary peptide in gingival crevicular fluid. [source]

The effect of hypoxia on pulmonary O2 uptake, leg blood flow and muscle deoxygenation during single-leg knee-extension exercise

EXPERIMENTAL PHYSIOLOGY, Issue 3 2004
Darren S. DeLorey
The effect of hypoxic breathing on pulmonary O2 uptake (VO2p), leg blood flow (LBF) and O2 delivery and deoxygenation of the vastus lateralis muscle was examined during constant-load single-leg knee-extension exercise. Seven subjects (24 ± 4 years; mean ±s.d.) performed two transitions from unloaded to moderate-intensity exercise (21 W) under normoxic and hypoxic (PETO2= 60 mmHg) conditions. Breath-by-breath VO2p and beat-by-beat femoral artery mean blood velocity (MBV) were measured by mass spectrometer and volume turbine and Doppler ultrasound (VingMed, CFM 750), respectively. Deoxy-(HHb), oxy-, and total haemoglobin/myoglobin were measured continuously by near-infrared spectroscopy (NIRS; Hamamatsu NIRO-300). VO2p data were filtered and averaged to 5 s bins at 20, 40, 60, 120, 180 and 300 s. MBV data were filtered and averaged to 2 s bins (1 contraction cycle). LBF was calculated for each contraction cycle and averaged to 5 s bins at 20, 40, 60, 120, 180 and 300 s. VO2p was significantly lower in hypoxia throughout the period of 20, 40, 60 and 120 s of the exercise on-transient. LBF (l min,1) was approximately 35% higher (P > 0.05) in hypoxia during the on-transient and steady-state of KE exercise, resulting in a similar leg O2 delivery in hypoxia and normoxia. Local muscle deoxygenation (HHb) was similar in hypoxia and normoxia. These results suggest that factors other than O2 delivery, possibly the diffusion of O2, were responsible for the lower O2 uptake during the exercise on-transient in hypoxia. [source]

Mass spectrometric detection of tyrosine sulfation in human pancreatic trypsinogens, but not in tumor-associated trypsinogen

FEBS JOURNAL, Issue 2 2008
Outi Itkonen
Trypsinogen-1 and -2 are well-characterized enzymes that are expressed in the pancreas and also in several other tissues. Many cancers produce trypsinogen isoenzymes that differ from the pancreatic ones with respect to substrate specificity and isoelectric point. These tumor-associated trypsinogens play a pivotal role in cancer progression and metastasis. The differences between these and the pancreatic isoenzymes have been suggested to be caused by post-translational modification, either sulfation or phosphorylation of a tyrosine residue. We aimed to elucidate the cause of these differences. We isolated trypsinogens from pancreatic juice and conditioned medium from a colon carcinoma cell line. Intact proteins, and tryptic and chymotryptic peptides were characterized by electrospray ionization mass spectrometry. We also used immunoblotting with antibody against phosphotyrosine and N-terminal sequencing. The results show that pancreatic trypsinogen-1 and -2 are sulfated at Tyr154, whereas tumor-associated trypsinogen-2 is not. Detachment of a labile sulfogroup could be demonstrated by both in-source dissociation and low-energy collision-induced dissociation in a tandem mass spectrometer. Tyrosine sulfation is an ubiquitous protein modification occurring in the secretory pathway, but its significance is often underestimated due to difficulties in its analysis. Sulfation is an almost irreversible modification that is thought to regulate protein,protein interactions and the activity of proteolytic enzymes. We conclude that the previously known differences in charge, substrate specificity and inhibitor binding between pancreatic and tumor-associated trypsinogens are probably caused by sulfation of Tyr154 in pancreatic trypsinogens. [source]

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

FEBS JOURNAL, Issue 2 2000
Søren Persson
We have used matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for mass determination of pigments and proteins in chlorosomes, the light-harvesting organelles from the photosynthetic green sulfur bacterium Chlorobium tepidum. By applying a small volume (1 µL) of a concentrated suspension of isolated chlorosomes directly to the target of the mass spectrometer we have been able to detect bacteriochlorophyll a and all the major homologs of bacteriochlorophyll c. The peak heights of the different bacteriochlorophyll c homologs in the MALDI spectra were proportional to peak areas obtained from HPLC analysis of the same sample. The same result was also obtained when whole cells of Chl. tepidum were applied to the target, indicating that MALDI-MS can provide a rapid method for obtaining a semiquantitative determination or finger-print of the bacteriochlorophyll homologs in a small amount of green bacterial cells. In addition to information on pigments, the MALDI spectra also contained peaks from chlorosome proteins. Thus we have been able with high precision to confirm the molecular masses of the chlorosome proteins CsmA and CsmE which have been previously determined by conventional biochemical and genetic methods, and demonstrate the presence of truncated versions of CsmA and CsmB. [source]

Influence of carbonation on aroma release from liquid systems using an artificial throat and a proton transfer reaction,mass spectrometric technique (PTR,MS)

FLAVOUR AND FRAGRANCE JOURNAL, Issue 5 2009
Maria Ángeles Pozo-Bayón
Abstract To determine whether carbonation affects aroma release from liquid systems, carbonated and non-carbonated flavoured model systems were prepared and volatile release was determined under static (equilibrium) and dynamic conditions. A model flavour system was added as a single compound or as a mixture of the six aroma compounds used in this study. Volatile release under dynamic conditions involved using a home-made device simulating an artificial throat, coupled to a proton transfer mass spectrometer (PTR,MS). The results showed that carbonation increased the release of most of the aroma compounds in both static and in dynamic testing conditions. The extent of this effect depended, however, on the physicochemical characteristics of the aroma compounds (the most volatile and most hydrophobic compounds were affected more). Release was also increased if the aroma compounds were added as a mixture rather than as individual compounds. CO2 appears to be a key factor responsible for the enhanced release of flavourings from carbonated liquid systems. Copyright © 2009 John Wiley & Sons, Ltd. [source]

Solid-phase aroma concentrate extraction (SPACEÔ ): a new headspace technique for more sensitive analysis of volatiles

FLAVOUR AND FRAGRANCE JOURNAL, Issue 3 2004
Masashi Ishikawa
Abstract The SPACEÔ (solid-phase aroma concentrate extraction) method is a modi,ed version of the SPME (solid-phase micro extraction) technique for headspace analysis, with increased area of the adsorbent to enable more sensitive analysis of volatiles. The SPACEÔ rod used in the technique is fabricated from stainless steel coated with an adsorbent mixture, consisting mainly of a graphite carbon. Initially, the SPACEÔ rod is ,xed in the head of a closed ,ask, where it adsorbs the aroma. Next, the rod is thermally desorbed on-line with a high-resolution gas chromatography,mass spectrometer (HRGC,MS). In the present experiments, SPACEÔ sampling reproducibility was determined by analysing a standard mixture and roasted coffee beans. The SPACEÔ rod collected the analytes with good reproducibility, with the exception of high polar compounds. Similar analyses of coffee powder were performed by SPME and other methods for comparison with the SPACEÔ method. The SPACEÔ method proved to have superior capabilities with high concentrations, and it produced a well-balanced chromatogram. Copyright © 2004 John Wiley & Sons, Ltd. [source]

Opportunities for ultra-high resolution analysis of essential oils using comprehensive two-dimensional gas chromatography: a review

FLAVOUR AND FRAGRANCE JOURNAL, Issue 3 2003
Robert Shellie
Abstract In comprehensive 2D gas chromatography, the entire sample is simultaneously subjected to analysis on two capillary columns. By using a suitable modulation interface between the primary and secondary columns, hundreds of fast, second-dimension chromatograms are produced. The data from these chromatograms are treated such that a 3D surface plot or a 2D contour plot of the components' individual retention times, on each column, as well as peak responses, are represented. In a properly tuned comprehensive 2D chromatogram, the individual sample components are spread throughout a 2D separation space, providing a signi,cant increase in the probability of resolving a greater number of sample components without increasing the analysis time. Comprehensive 2D,GC has proved useful for high-resolution conventional essential oil analysis as well as high-resolution enantioselective essential oil analysis. Combining comprehensive 2D,GC with either a quadrupole or time-of-,ight mass spectrometer gives a powerful 3D analysis technique, which is extremely effective for complex sample analysis. The present status and opportunities arising from these ultra-high resolution approaches are discussed herein. Copyright © 2003 John Wiley & Sons, Ltd. [source]

Real-time quadrupole mass spectrometer analysis of gas in borehole fluid samples acquired using the U-tube sampling methodology

GEOFLUIDS (ELECTRONIC), Issue 3 2006
B. M. FREIFELD
Abstract Sampling of fluids in deep boreholes is challenging because of the necessity of minimizing external contamination and maintaining sample integrity during recovery. The U-tube sampling methodology was developed to collect large volume, multiphase samples at in situ pressures. As a permanent or semi-permanent installation, the U-tube can be used for rapidly acquiring multiple samples or it may be installed for long-term monitoring applications. The U-tube was first deployed in Liberty County, TX to monitor crosswell CO2 injection as part of the Frio CO2 sequestration experiment. Analysis of gases (dissolved or separate phase) was performed in the field using a quadrupole mass spectrometer, which served as the basis for determining the arrival of the CO2 plume. The presence of oxygen and argon in elevated concentrations, along with reduced methane concentration, indicates sample alteration caused by the introduction of surface fluids during borehole completion. Despite producing the well to eliminate non-native fluids, measurements demonstrate that contamination persists until the immiscible CO2 injection swept formation fluid into the observation wellbore. [source]

Preparation of a Synthetic Titanite Glass Calibration Material for In Situ Microanalysis by Direct Fusion in Graphite Electrodes: A Preliminary Characterisation by EPMA and LA-ICP-MS

GEOSTANDARDS & GEOANALYTICAL RESEARCH, Issue 2 2005
Magne Ødegård
matériaux de calibration; microanalyse; fusion directe; électrodes de graphite; verre de titanite This paper describes a technique for the preparation of a titanite (CaTiSiO5) glass calibration material for use in in situ microanalysis of major, minor, and trace elements in geological materials. The starting composition was a titanite matrix doped with minor and trace elements at , 200 ,g g -1. The elements Sc, Y, REEs, Th and U were added in the form of nitrates in solution, and the elements V, Cr, Mn, Fe, Co, Ni, Zr, Nb, Hf and W were added as solid oxides. The synthetic titanite glass was produced by direct fusion by resistance heating in graphite electrodes at 1600-1700 °C, and quenched in air. Backscattered electron images indicate good homogeneity, with no signs of separate phases or vesicles, and analysis of the major elements Ca, Ti and Si by electron microprobe showed relative standard deviations between 0.5 and 0.7%, based on six independent measurements. Deviations from nominal concentrations for Ca, Si and Ti were measured to -1.2, -3.3 and -0.8%, respectively. The homogeneity of the trace elements in the glass was assessed by LA-ICP-MS analyses, using NIST SRM 610, 612 and 616 as external calibrators, and Ca as the internal standard element. Determinations were made both with a quadrupole mass spectrometer and a sector field instrument, and both raster and spot modes of analysis were used. For the majority of doped elements, precision was better than 10%, and relative deviations from nominal values were, with few exceptions, between 5 and 10%. Cet article décrit une technique de préparation d'un verre de composition CaTiSiO5 (titanite) pour l'utiliser comme matériau de calibration lors de microanalyses in situ des éléments majeurs, mineurs et en trace dans des matériaux géologiques. La composition de départ a une matrice de titanite, dopée avec des éléments mineurs et en trace à une concentration de , 200 ,g g-1. Les éléments Sc, Y, REE, Th et U ont été ajoutés sous forme de nitrates en solution et les éléments V, Cr, Mn, Fe, Co, Ni, Zr, Nb, Hf et W sous forme d'oxydes solides. Le verre synthétique de titanite a été produit par fusion directe avec un chauffage par des résistances dans des électrodes de graphite à 1600-1700 °C suivi d'un refroidissement rapide à l'air. Les images obtenues par électrons rétrodiffusés montrent que le verre présente une bonne homogénéité, sans aucun signe de phases individualisées ou de vésicules, et l'analyse des éléments majeurs Ca, Ti et Si par microsonde électronique a des déviations standard relatives (RSD) entre 0.5 et 0.7% provenant de six mesures indépendantes. Les déviations par rapport aux concentrations calculées, pour Ca, Si et Ti, sont de -1.2, -3.3 et -0.8% respectivement. L'homogénéité de répartition des éléments en trace dans le verre a été vérifiée par des analyses LA-ICP-MS, en utilisant les matériaux de référence NIST SRM 610, 612 et 616 pour la calibration externe et Ca comme élément standard interne. Les déterminations ont été faites avec un spectromètre de masse de type quadrupôle et un autre de type secteur magnétique, par des analyses à la fois en mode balayage et en mode ponctuel. Pour la majorité des éléments dopés, la précision est meilleure que 10% et les déviations standard relatives par rapport aux valeurs calculées sont, à quelques exceptions près, entre 5 et 10%. [source]

A kinetic and product study of reaction of chlorine atom with CH3CH2OD

INTERNATIONAL JOURNAL OF CHEMICAL KINETICS, Issue 11 2004
Mary A. Crawford
The reaction of atomic chlorine with CH3CH2OD has been examined using a discharge fast flow system coupled to a mass spectrometer combined with the relative rate method (RR/DF/MS). At 298 ± 2 K, the rate constant for the Cl + CH3CH2OD reaction was determined using cyclohexane as a reference and found to be k3 = (1.13 ± 0.21) × 10,10 cm3 molecule,1 s,1. Mass spectral studies of the reaction products resulted in yields greater than 97% for the combined hydrogen abstraction at the , and , sites (3a + 3b) and less than 3% at the hydroxyl site (3c). As a calibration of the apparatus and the RR/DF/MS technique, the rate constant of the Cl + CH3CH2OH reaction was also determined using cyclohexane as the reference, and a value of k2 = (1.05 ± 0.07) × 10,10 cm3 molecule,1 s,1 was obtained at 298 ± 2 K, which was in excellent agreement with the value given in current literature. © 2004 Wiley Periodicals, Inc. Int J Chem Kinet 36: 584,590, 2004 [source]

Desorption kinetics of model polar stratospheric cloud films measured using Fourier Transform Infrared Spectroscopy and Temperature-Programmed Desorption

INTERNATIONAL JOURNAL OF CHEMICAL KINETICS, Issue 5 2001
Birgit G. Koehler
This study combines Fourier transform infrared (FTIR) spectroscopy and temperature-programmed desorption to examine the evaporation kinetics of thin films of crystalline nitric acid hydrates, solid amorphous H2O/HNO3 mixtures, H2O,ice, ice coated with HCl, and solid HNO3. IR spectroscopy measured the thickness of each film as it evaporated, either at constant temperature or during a linear temperature ramp (temperature-programmed infrared, TPIR). Simultaneously, a mass spectrometer measured the rate of evaporation directly by monitoring the evolution of the molecules into the gas phase (temperature-programmed desorption, TPD). Both TPIR and TPD data provide a measurement of the desorption rate and yield the activation energy and preexponential factor for desorption. TPD measurements have the advantage of producing many data points but are subject to interference from experimental difficulties such as uneven heating from the edge of a sample and sample-support as well as pumping-speed limitations. TPIR experiments give clean but fewer data points. Evaporation occurred between 170 and 215 K for the various films. Ice evaporates with an activation energy of 12.9 ± 1 kcal/mol and a preexponential factor of 1 × 1032±1.5 molec/cm2 s, in good agreement with the literature. The beta form of nitric acid trihydrate, ,,NAT, has an Edes of 15.6 ± 2 kcal/mol with log A = 34.3 ± 2.3; the alpha form of nitric acid trihydrate, ,,NAT, is around 17.7 ± 3 kcal/mol with log A = 37.2 ± 4. For nitric acid dihydrate, NAD, Edes is 17.3 ± 2 kcal/mol with log A = 35.9 ± 2.6; for nitric acid monohydrate, NAM, Edes is 13 ± 3 kcal/mol with log A = 31.4 ± 3. The ,,NAT converts to ,,NAT during evaporation, and the amorphous solid H2O/HNO3 mixtures crystallize during evaporation. The barrier to evaporation for pure nitric acid is 14.6 ± 3 kcal/mol with log A = 34.4 ± 3. © 2001 John Wiley & Sons, Inc. Int J Chem Kinet 33: 295,309, 2001 [source]

Dichlorvos, chlorpyrifos oxon and Aldicarb adducts of butyrylcholinesterase, detected by mass spectrometry in human plasma following deliberate overdose

JOURNAL OF APPLIED TOXICOLOGY, Issue 6 2010
Bin Li
Abstract The goal of this study was to develop a method to detect pesticide adducts in tryptic digests of butyrylcholinesterase in human plasma from patients poisoned by pesticides. Adducts to butyrylcholinesterase in human serum may serve as biomarkers of pesticide exposure because organophosphorus and carbamate pesticides make a covalent bond with the active site serine of butyrylcholinesterase. Serum samples from five attempted suicides (with dichlorvos, Aldicarb, Baygon and an unknown pesticide) and from one patient who accidentally inhaled dichlorvos were analyzed. Butyrylcholinesterase was purified from 2 ml serum by ion exchange chromatography at pH 4, followed by procainamide affinity chromatography at pH 7. The purified butyrylcholinesterase was denatured, digested with trypsin and the modified peptide isolated by HPLC. The purified peptide was analyzed by multiple reaction monitoring in a QTRAP 4000 mass spectrometer. This method successfully identified the pesticide-adducted butyrylcholinesterase peptide in four patients whose butyrylcholinesterase was inhibited 60,84%, but not in two patients whose inhibition levels were 8 and 22%. It is expected that low inhibition levels will require analysis of larger serum plasma volumes. In conclusion, a mass spectrometry method for identification of exposure to live toxic pesticides has been developed, based on identification of pesticide adducts on the active site serine of human butyrylcholinesterase. Copyright © 2010 John Wiley & Sons, Ltd. [source]

High-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry for the determination of flocoumafen and brodifacoum in whole blood

JOURNAL OF APPLIED TOXICOLOGY, Issue 1 2007
Mi-cong Jin
Abstract A high-performance liquid chromatographic,tandem mass spectrometric (HPLC,MS,MS) assay was developed and validated to determine quantitatively flocoumafen and brodifacoum in whole blood using warfarin as an internal standard (IS). Liquid,liquid extraction, using ethyl acetate, was used to isolate flocoumafen, brodifacoum and the IS from the biological matrix. Detection was performed on a mass spectrometer by negative electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The calibration curves were linear (r2 > 0.998) in the concentration range of 0.1,100.0 ng ml,1 with a lower limit of quantification of 0.05 ng ml,1 for flocoumafen, and 0.1 ng ml,1 for brodifacoum in whole blood. Intra-day and inter-day relative standard deviations (RSDs) were less than 8.0% and 10.8%, respectively. Recoveries of flocoumafen and brodifacoum ranged from 78.0% to 83.7%. This assay can be used to determine trace flocoumafen and brodifacoum in whole blood to investigate suspected poisoning of human and animals. Copyright © 2006 John Wiley & Sons, Ltd. [source]