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Mass Spectrometric Detection (mass + spectrometric_detection)
Kinds of Mass Spectrometric Detection Selected AbstractsAnalysis of allergens in metalworking fluidsCONTACT DERMATITIS, Issue 5 2008Maj-Len Henriks-Eckerman Background:, Metalworking fluids (MWFs) are well-known causes of occupational contact dermatitis in machinists. Objective:, To gain information about skin sensitizers in MWFs and to compare it with the information in safety data sheets (SDSs). Methods:, A total of 17 samples of MWF concentrates were analysed for skin sensitizers known or suspected to be used in MWF. Alkanolamines, formaldehyde, isothiazolinones, methyldibromo glutaronitrile (MDBGN), and iodopropynyl butylcarbamate (IPBC) were separated by liquid chromatography. Resin acids of colophonium (colophony) were separated by gas chromatography. The substances were identified with mass spectrometric detection and ultraviolet detection. Results:, Of the MWFs, 15 contained 6,39% of alkanolamines, mostly monoethanolamine and triethanolamine. Formaldehyde was detected in all MWFs: the concentrations of total formaldehyde ranged between 0.002% and 1.3%. Benzisothiazolinone and octylisothiazolinone were detected in one fluid each. IPBC was detected in nine MWFs, and the highest concentration was 0.09%. Methylisothiazolinone and MDBGN were not detected in any of the fluids. Resin acids of colophonium were detected in seven MWFs in concentrations ranging from 0.41% to 3.8%. On the whole, the allergens analysed were poorly declared in the SDSs. Conclusions:, The content of total formaldehyde was not declared in any SDS. IPBC, a relatively new allergen, seems to be common in MWFs. Isothiazolinones may be relevant allergens of machinists, and they should be analysed in MWFs in case other sources are not identified. The occupational relevance of positive patch test results to MWF ingredients in machinists is difficult to determine if information in the SDSs is relied upon. [source] Historical review of sample preparation for chromatographic bioanalysis: pros and consDRUG DEVELOPMENT RESEARCH, Issue 3 2007Min S. Chang Abstract Sample preparation is a major task in a regulated bioanalytical laboratory. The sample preparation procedure significantly impacts assay throughput, data quality, analysis cost, and employee satisfaction. Therefore, selecting and optimizing an appropriate sample preparation method is essential for successful method development. Because of our recent expertise, this article is focused on sample preparation for high-performance liquid chromatography with mass spectrometric detection. Liquid chromatography with mass spectrometric detection (LC-MS) is the most common detection technique for small molecules used in regulated bioanalytical laboratories. The sample preparation technologies discussed are pre-extraction and post-extraction sample processing, protein precipitation (PPT), liquid,liquid extraction (LLE), offline solid-phase extraction (SPE), and online solid-phase extraction. Since all these techniques were in use for more than two decades, numerous applications and variations exist for each technique. We will not attempt to categorize each variation. Rather, the development history, a brief theoretical background, and selected references are presented. The strengths and the limitations of each method are discussed, including the throughput improvement potential. If available, illustrations from presentations at various meetings by our laboratory are used to clarify our opinion. Drug Dev Res 68:107,133, 2007. ©2007 Wiley-Liss, Inc. [source] Cyclodextrin-based nonaqueous electrokinetic chromatography with UV and mass spectrometric detection: Application to the impurity profiling of amiodarone,ELECTROPHORESIS, Issue 17 2008Roelof Mol Abstract The potential of nonaqueous electrokinetic chromatography (NAEKC) using cyclodextrins (CD) for the analysis of basic drugs and related compounds was evaluated. Both UV absorbance and mass spectrometric (MS) detection were employed. Addition of neutral CD to the NA background electrolyte did not significantly enhance the separation of a test mixture of basic drugs, and no change in selectivity was observed. In contrast, anionic single-isomer-sulfated CD strongly added to the selectivity of the NAEKC system inducing an improved resolution among the test compounds and increasing the migration time window. The applicability of the NAEKC system using anionic CD is demonstrated by the profiling of a sample of the drug amiodarone that had been stored for 1,year at room temperature. Amiodarone is poorly soluble in water. NAEKC-UV analysis indicated the presence of at least seven impurities in the amiodarone sample. In order to identify these compounds, the NAEKC system was coupled directly to electrospray ionization (ESI) ion-trap MS. The total of detected impurities increased to 12 due to the added sensitivity and selectivity of MS detection. Based on the acquired MS/MS data, three sample constituents could be identified as ,known' impurities (British Pharmacopoeia), whereas for three unknown impurities molecular structures could be proposed. Estimated limits of detection for amiodarone using the NAEKC method were 1,,g/mL with UV detection and 15,ng/mL with ESI-MS detection (full-scan). Based on relative responses, the impurity content of the stored drug substance was estimated to be 0.33 and 0.47% using NAEKC-UV and NAEKC-ESI-MS, respectively. [source] A multilayer poly(dimethylsiloxane) electrospray ionization emitter for sample injection and online mass spectrometric detectionELECTROPHORESIS, Issue 24 2005Jamie M. Iannacone Abstract An ESI emitter made of poly(dimethylsiloxane) interfaces on-chip sample preparation with MS detection. The unique multilayer design allows both the analyte and the spray solutions to reside on the device simultaneously in discrete microfluidic environments that are spatially separated by a polycarbonate track-etched, nanocapillary array membrane (NCAM). In direct spray mode, voltage is applied to the microchannel containing a spray solution delivered via a syringe pump. For injection, the spray potential is lowered and a voltage is applied that forward biases the membrane and permits the analyte to enter the spray channel. Once the injection is complete, the bias potential is switched off, and the spray voltage is increased to generate the ESI of the injected analyte plug. Consecutive injections of a 10,,M bovine insulin solution are reproducible and produce sample plugs with limited band broadening and high quality mass spectra. Peptide signals are observed following transport through the NCAM, even when the peptide is dissolved in solutions containing up to 20% seawater. The multilayer emitter shows great potential for performing multidimensional chemical manipulations on-chip, followed by direct ESI with negligible dead volume for online MS analysis. [source] Determination of bupivacaine and metabolites in rat urine using capillary electrophoresis with mass spectrometric detectionELECTROPHORESIS, Issue 14 2003Ryan M. Krisko Abstract A method using capillary electrophoresis-mass spectrometry (CE-MS) was developed for the structural elucidation of bupivacaine and metabolites in rat urine. Prior to CE-MS analysis, solid-phase extraction (SPE) was used for sample cleanup and preconcentration purposes. Exact mass and tandem mass spectrometric (MS/MS) experiments were performed to obtain structural information about the unknown metabolites. Two instruments with different mass analyzers were used for mass spectrometric detection. A quadrupole time-of-flight (Q-TOF) and a magnetic sector hybrid instrument were coupled to CE and used for the analysis of urine extracts. Hydroxybupivacaine as well as five other isomerically different metabolites were detected including methoxylated bupivacaine. [source] Recent progress in the development, characterization and application of polymeric pseudophases for electrokinetic chromatographyELECTROPHORESIS, Issue 22-23 2002Christopher P. Palmer Abstract This review article details the development, characterization and application of polymeric materials as pseudostationary phases for electrokinetic chromatography over the past two years. Recent developments in cationic polymers and anionic siloxane, acrylamide and polymerized surfactants (micelle polymers) are reviewed. Also reviewed is recent progress in the development and characterization of chiral polymeric phases for chiral separations by electrokinetic chromatography, and application of a polymeric pseudophase with electrospray ionization mass spectrometric detection. [source] Impact of changes in analytical techniques for the measurement of polychlorinated biphenyls and organochlorine pesticides on temporal trends in herring gull eggsENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 7 2010Shane R. de Solla Abstract Changes in analytical approaches during the tenure of monitoring programs for organochlorine (OC) pesticides and polychlorinated biphenyls (PCBs) may affect estimates of temporal trends. We used an in-house reference material to create multiplication factors to adjust the estimates of OC pesticides and PCBs (Aroclor equivalents) in Great Lake herring gull eggs analyzed using electron capture detection (1987,1997) to be more equivalent to estimates using mass spectrometric detection (1998,2005) as well as accompanying differences in analytical procedures. We examined temporal trends in contaminant concentrations in herring gull eggs using change point regressions, to determine whether significant changes in long-term trends were associated with analytical methodology. The highest frequency of change point occurrences shifted from 1997 (when analytical methodology was altered) to 2003 after data adjustment. The explanatory power (r2) of the regressions was lower after adjustment, although only marginally so (mean r2 difference,=,0.04). The initial rates of decline before change points in contaminant concentrations were generally slower after the data adjustment, but after any change points the declines were not significantly different. The regression models did not change for 83.3% of the cases. The effects on the interpretation of long-term temporal trends in herring gull eggs, although not negligible, were minor relative to the magnitude of the temporal changes. Environ. Toxicol. Chem. 2010;29:1476,1483. © 2010 SETAC [source] Determination of phthalate esters in cosmetics by gas chromatography with flame ionization detection and mass spectrometric detectionINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 4 2005Huiming Chen GC-FID; GC-MS; Produits cosmétiques; Esters Phtaliques Synopsis A gas chromatography coupled with flame ionization detection (GC-FID) and mass spectrometric detection (MSD) method was developed to determine the six kinds of phthalate esters [dimethyl phthalate (DMP), diethyl phthalate (DEP), di- n -butyl phthalate (DBP), benzyl butyl phthalate (BBP), di(2-ethylhexyl) phthalate (DEHP) and di- n -octyl phthalate (DOP)] in cosmetics (solid, cream and liquid cosmetics). The cosmetics were extracted with methanol by ultrasonic and then separated with high-speed centrifugation. The upper clear layer was dried and filtered through a 0.45 ,m pore diameter filter. The filtrate was injected into GC-FID/GC-MS for detection. GC-FID chromatogram was applied for qualitative analysis, external standard method was used for quantitative analysis. Confirmation of phthalate presence was undertaken by GC-EI-MS. The recovery range of all phthalates were between 92.0 and 110.0% with relative standard deviations between 1.95 and 5.92%. The low detection limits of the method were: 0.1 ng for DMP, DEP, DBP and BBP, 0.5 ng for DEHP and DOP. The method had advantages of high precision and sensitivity, simplicity of pretreatment. The method can be used to test the six kinds of phthalate esters in cosmetics. Resume Une méthode d'analyse par chromatographie gazeuse couplée à une détection par ionization de flamme (GC - FID) et une détection spectrométrique de masse (MSD) a été développée pour analyser 6 sortes d'esters phtaliques (phtalate de diméthyle (DMP), phtalate de diéthyle (DEP), phtalate de di- n -butyle (DBP), phtalate de benzylbutyle (BBP), phtalate de di-2-éthylhexyle (DEHP) et phtalate de di- n -octyle (DOP)) dans des produits cosmétiques (solides, crèmes et liquides). Les produits cosmétiques sont extraits au méthanol sous ultrason, puis séparés par ultracentrifugation. La phase supérieure limpide est déshydratée et filtrée sur un filtre de diamètre de pore moyen égal à 0,45 ,m. Le filtrat est injecté dans le système GC - FID/GC-MS pour analyse. Les chromatogrammes GC-FID sont utilisés pour l'analyse qualitative, des standards externes ont été utilisés pour l'analyse quantitative. La GC-EI-MS permet de confirmer la présence des esters phtaliques. Le taux de récupération de tous les esters est compris entre 92 et 110% avec une déviation standard allant de 1,95%à 5,92%. La limite de détection par cette méthode est de 0,1 ng pour DMP, DEP, DBP et BBP, 0,5 ng pour DEHP et DOP. Les avantages de cette méthode sont sa haute précision, sa sensibilité et la simplicité du prétraitement. Cette méthode peut être utilisée pour doser la présence des six sortes d'esters phtaliques dans des produits cosmétiques. [source] An algorithm for thorough background subtraction from high-resolution LC/MS data: application for detection of glutathione-trapped reactive metabolitesJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 9 2008Haiying Zhang Abstract A control sample background-subtraction algorithm was developed for thorough subtraction of background and matrix-related signals in high-resolution, accurate mass liquid chromatography/mass spectrometry (LC/MS) data to reveal ions of interest in an analyte sample. This algorithm checked all ions in the control scans within a specified time window around the analyte scan for potential subtraction of ions found in that analyte scan. Applying this method, chromatographic fluctuations between runs were dealt with and background and matrix-related signals in the sample could be thoroughly subtracted. The effectiveness of this algorithm was demonstrated using four test compounds, clozapine, diclofenac, imipramine, and tacrine, to reveal glutathione (GSH)-trapped reactive metabolites after incubation with human liver microsomes supplemented with GSH (30 µM compound, 45-min incubation). Using this algorithm with a ± 1.0 min control scan time window, a ± 5 ppm mass error tolerance, and appropriate control samples, the GSH-trapped metabolites were revealed as the major peaks in the processed LC/MS profiles. Such profiles allowed for comprehensive and reliable identification of these metabolites without the need for any presumptions regarding their behavior or properties with respect to mass spectrometric detection. The algorithm was shown to provide superior results when compared to several commercially available background-subtraction algorithms. Many of the metabolites detected were doubly charged species which would be difficult to detect with traditional GSH adduct screening techniques, and thus, some of the adducts have not previously been reported in the literature. Copyright © 2008 John Wiley & Sons, Ltd. [source] Determination of clavulanic acid in calf plasma by liquid chromatography tandem mass spectrometryJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 11 2006Tim Reyns Abstract A method for the quantification of clavulanic acid in calf plasma using high-performance liquid chromatography combined with electrospray ionization (ESI) mass spectrometry, operating in the negative ionization mode (LC-MS/MS), is presented. Sample preparation includes a simple and fast deproteinization with acetonitrile and a back-extraction of the acetonitrile with dichloromethane. Chromatography is performed on a reversed-phase PLRP-S polymeric column using 0.05% formic acid in water and acetonitrile. The limit of quantification is 25 ng/ml, which is lower than other published methods using ultraviolet (UV), fluorimetric or mass spectrometric detection. The limit of detection is calculated to be 3.5 ng/ml. The stability of clavulanic acid was demonstrated according to The Guidelines of Bioanalytical Method Validation of The Food and Drug Administration (FDA): freeze and thaw stability, short-term stability, long-term stability, stock solution stability and postpreparative stability. The method is used in a pharmacokinetic and bioequivalence study of amoxycillin/clavulanic acid formulations in calves. Copyright © 2006 John Wiley & Sons, Ltd. [source] Quantitative analysis of the P-glycoprotein inhibitor Elacridar (GF120918) in human and dog plasma using liquid chromatography with tandem mass spectrometric detectionJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 10 2004Ellen Stokvis Abstract A liquid chromatographic/tandem mass spectrometric (LC/MS/MS) method for the determination of the P-glycoprotein and breast cancer resistance protein inhibitor Elacridar in human and dog plasma is described. The internal standard was stable isotopically labelled Elacridar. Sample pretreatment involved liquid,liquid extraction with tert -butyl methyl ether. Analysis of Elacridar and internal standard was performed by reversed-phase LC on a basic stable minibore analytical column with an eluent consisting of acetonitrile and aqueous ammonia. An API-2000 triple-quadrupole mass spectrometer with an electrospray ion source was used in the positive-ion multiple reaction monitoring mode. The run time per sample was only 6 min. The method is sensitive and specific, with a dynamic range from 1 to 500 ng ml,1 from 100 µl of human or dog plasma. The accuracy of the method was within 15% bias and the precision was lower than 15% for all tested concentration levels and in both matrices. The method is simple and the liquid,liquid extraction produces clean samples. This method was successfully applied to support the pharmacokinetics of a clinical trial in which orally applied Elacridar was used as a bioavailability enhancer. Copyright © 2004 John Wiley & Sons, Ltd. [source] Direct-temperature mass spectrometric detection of volatile terpenoids and natural terpenoid polymersin fresh and artificially aged resinsJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 6 2003Dominique Scalarone Abstract Electron impact (EI) ionization and ammonia chemical ionization (NH3/CI) direct-temperature mass spectrometry (DTMS) was used to characterize five natural terpenoid resins: dammar, mastic, colophony, Manila copal and sandarac. Compositional differences were highlighted by the identification of low molecular mass compounds, ranging from di- to triterpenoids, and polymeric components, based on polycadinene and polycommunic acid. Photo-ageing processes occurring under accelerated indoor and outdoor exposure conditions were also investigated. NH3/CI and tetramethylammonium hydroxide EI were applied to increase the sensitivity towards highly oxidized molecules. Oxidation and cross-linking reactions were found to affect mostly triterpenoid resins and diterpenoid abietane and pimarane molecules. Oxidation proceeds through a radical mechanism, generally starting from conjugated double bonds. Oxygen atoms are incorporated in the terpenoid structures in the form of alcohols, ketones and carboxylic acids. Oxidized cadinene oligomers released by pyrolytic degradation of the polycadinene fraction of dammar were detected even in unaged samples. Evidence is given indicating the occurrence of cleavages in the cross-linked polycommunic structure of aged sandarac and Manila copal. Bond scissions produce oligomeric fragments based on the communic acid structure and sufficiently volatile to be desorbed at low temperature in DTMS measurements. Copyright © 2003 John Wiley & Sons, Ltd. [source] Capillary electrophoresis of intact basic proteins using noncovalently triple-layer coated capillariesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 14 2009Rob Haselberg Abstract The usefulness of a noncovalent, positively charged capillary coating for the efficient analysis of intact basic proteins with CE was studied. Capillaries were coated by subsequent flushing with solutions of 10% w/v Polybrene (PB), 3% w/v dextran sulfate (DS), and again 10% w/v PB. Coating characterization studies showed that stable coatings could be produced which exhibited a pH-independent and highly reproducible EOF. The PB,DS,PB coating was evaluated with Tris phosphate BGEs of various pH using the four basic model proteins: ,-chymotrypsinogen A, ribonuclease A, cytochrome c, and lysozyme. Typical migration time RSDs for the proteins were less than 0.85%, and apparent plate numbers were above 125 000 using a capillary length of 40 cm. The high separation efficiency allowed detection of several minor impurities in the model proteins. Using a BGE of medium pH, the CE system with triple-layer coating appeared to be useful for the repeatable profiling of recombinant humanized mouse monoclonal immunoglobulin G1 showing a characteristic pattern of glycoforms. The CE system was also applied to the characterization of two llama antibodies, which were produced in Saccharomyces cerevisiae, revealing the presence of a side product in one of the antibodies. The high migration time stability allowed the reliable determination of antibody,antigen binding by monitoring migration time shifts. Finally, the feasibility of using the PB,DS,PB coated capillaries for CE with mass spectrometric detection was shown by the characterization of the impure llama antibody sample. [source] A novel HS-SBSE system coupled with gas chromatography and mass spectrometry for the analysis of organochlorine pesticides in water samplesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 20 2008Paula Grossi Abstract A methodology to analyze organochlorine pesticides (OCPs) in water samples has been accomplished by using headspace stir bar sorptive extraction (HS-SBSE). The bars were in house coated with a thick film of PDMS in order to properly work in the headspace mode. Sampling was done by a novel HS-SBSE system whereas the analysis was performed by capillary GC coupled mass spectrometric detection (HS-SBSE-GC-MS). The extraction optimization, using different experimental parameters has been established by a standard equilibrium time of 120 min at 85°C. A mixture of ACN/toluene as back extraction solvent promoted a good performance to remove the OCPs sorbed in the bar. Reproducibility between 2.1 and 14.8% and linearity between 0.96 and 1.0 were obtained for pesticides spiked in a linear range between 5 and 17 ng/g in water samples during the bar evaluation. [source] Determination of the pesticides considered as endocrine-disrupting compounds (EDCs) by solid-phase extraction followed by gas chromatography with electron capture and mass spectrometric detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 12 2007Vasiliki I. Valsamaki Abstract An SPE method followed by GC-electron capture detection (ECD) with confirmation by MS for the trace determination of four pesticides considered as endocrine-disrupting compounds (EDCs) in natural waters and sediments has been developed. Target analytes, fenarimol, fenvalerate, pendimethalin, and vinclozolin, belong to different chemical groups and are used mainly in agriculture. In the present study, analysis employs an offline SPE step for the extraction of the target analytes from natural waters. Sonication and subsequent SPE clean-up was used for extraction and purification of the sediment samples which were finally treated with activated copper powder. The type of SPE disk, eluents as well as solution parameters including pH value, and concentrations of salts and humic substances were examined for the efficiency of the method. The recoveries of all pesticides were in relatively high levels, ranging from 75 to 97% for waters and 71 to 84% for sediment samples. Both methods were applied to real water and sediment samples and the presence of the tested compounds was investigated. [source] Simultaneous qualification and quantification of eight triterpenoids in Radix Achyranthis Bidentatae by high-performance liquid chromatography with evaporative light scattering detection and mass spectrometric detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 6 2007Juan Li Abstract An HPLC with evaporative light scattering detection (ELSD) and ESI-MS was established for the simultaneous determination of eight triterpenoids in Radix Achyranthis Bidentatae. The optimal chromatographic conditions were achieved on a Zorbax C18 column by linear gradient elution with 0.08% v/v aqueous formic acid and ACN as the mobile phase at the flow rate of 0.8 mL/min. Temperature for the detector drift tube was set at 101°C and the nitrogen flow rate was 2.8 L/min. The identities of the analytes were accomplished by comparing retention times and mass data with those of reference compounds. The validation of the method included tests of linearity, sensitivity, repeatability, recovery, and stability. All the calibration curves of the eight triterpenoids showed good linear regression (R2 >0.997) within the test ranges. The method provides desirable repeatability with overall intra- and interday variations of less than 4.9%. The obtained recoveries varied between 93.6 and 98.1% while the RSDs were below 3.9% (n = 3). The analysis results indicate that the content of investigated triterpenoids in Radix Achyranthis Bidentatae from different locations was greatly diverse, and the triterpenoids could be used as chemical markers for the discrimination of genuine and ungenuine crude drugs. [source] Separation of triacylglycerols in a complex lipidic matrix by using comprehensive two-dimensional liquid chromatography coupled with atmospheric pressure chemical ionization mass spectrometric detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 8 2006Paola Dugo Abstract The present investigation describes the employment of a comprehensive 2-D HPLC system, based on the combination of a silver ion and an RP column, for the characterization of the triacylglycerol (TAG) fraction of a very complex lipidic sample: donkey milk fat. The TAGs were grouped on the resulting bidimensional contour plot according to their double bond numbers (aligned along vertical bands) and according to their partition numbers (aligned along horizontal bands). Peak assignment was supported by using atmospheric pressure chemical ionization mass spectrometric (APCI-MS) detection. The combination of the enhanced resolving power of comprehensive multidimensional LC, the formation of ordered 2-D patterns, and APCI-MS detection proved to be an effective tool for the characterization of the complex matrix, enabling the separation and identification of nearly 60 TAGs. [source] Enantiomeric separation of mirtazapine and its metabolites by nano-liquid chromatography with UV-absorption and mass spectrometric detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 14 2005Salvatore Fanali Abstract Mirtazapine (MIR) and two of its main metabolites, namely, 8-hydroxymirtazapine and N -desmethylmirtazapine, were separated in totheir enantiomers by nanoLC in a laboratory-made fused-silica capillary column (75 ,m ID) packed with a vancomycin-modified silica stationary phase. The simultaneous separation of the three couples of the studied enantiomers was achieved in less than 33 min, using an experimentally optimized mobile phase delivered in the isocratic mode. Optimization of the mobile-phase composition was achieved by testing the influence of the buffer pH and concentration, the water concentration, the organic modifier type and concentration, and on the retention and resolution of the analytes. The optimum mobile-phase composition contained 500 mM ammonium acetate pH 4.5/water/MeOH/MeCN, 1 : 14 : 40 : 45 v/v/v/v. Using a UV detector at 205 nm, the method was validated studying several experimental parameters such as LOD and LOQ, intraday and interday repeatability, and linearity. Good results were achieved: LOD and LOQ were in the range 5,15 and 10,40 ,g/mL, respectively (the highest value was obtained for the DEMIR enantiomers); correlation coefficients, 0.9993,0.9999; the intraday and interday precision was acceptable (RSD < 2%) using an internal standard. The method was tested for the separation of the studied enantiomers in an extracted (solid-phase) serum sample spiked with standard racemic mixture of MIR and its two metabolites. Finally, the nanoLC system was connected to a mass spectrometer through a nanoelectrospray interface and the MS, MS2, and MS3 spectra were acquired showing the potential of the system used for characterization and identification of the separated analytes. [source] Validation of a quantitative assay using GC/MS for trace determination of free and conjugated estrogens in environmental water samplesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 1-2 2003Asmaa Mouatassim-Souali Abstract It has been shown that sewage effluent can discharge human hormones and pharmaceuticals, particularly estrogens and synthetic chemicals, which are able to disrupt animal and human endocrine systems into surface waters. Since many surface waters receive sewage effluent and are subsequently used to produce drinking water, it is of principal interest to assess their contamination level and thereby their possible public health and environmental impact. To date, no data concerning the occurrence of estrogens present in the French aquatic environment are available. We therefore developed and validated an analytical procedure, which allows simultaneous quantitative determination of three natural estrogens, 17,-estradiol, estriol, and estrone and one of the synthetic estrogens most widely used in contraception, ethinylestradiol, in water. Water samples are extracted using solid-phase extraction (SPE) and then separated by gas-chromatography coupled with mass spectrometric detection. Under our conditions, detection limits of estrogens reached the pg range of injected sample, i. e. less than 0.1 ng L,1. Conjugated estrogens were also investigated using the same procedure as described above but with a enzymatic hydrolysis preliminary step before extraction. The analysis of samples collected from four wastewater treatment plants and from surface water showed significant concentrations of estrogens ranging from 2 to 18 ng L,1 and from 0.5 to 3 ng L,1, respectively. Furthermore, no estrogen conjugated forms were detected in the water samples. [source] Acrylamide levels in Finnish foodstuffs analysed with liquid chromatography tandem mass spectrometryMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 2 2007Susanna Eerola Abstract Sample clean-up and HPLC with tandem mass spectrometric detection (LC-MS/MS) was validated for the routine analysis of acrylamide in various foodstuffs. The method used proved to be reliable and the detection limit for routine monitoring was sensitive enough for foods and drinks (38 ,g/kg for foods and 5 ,g/L for drinks). The RSDs for repeatability and day-to-day variation were below 15% in all food matrices. Two hundred and one samples which included more than 30 different types of food and foods manufactured and prepared in various ways were analysed. The main types of food analysed were potato and cereal-based foods, processed foods (pizza, minced beef meat, meat balls, chicken nuggets, potato-ham casserole and fried bacon) and coffee. Acrylamide was detected at levels, ranging from nondetectable to 1480 ,g/kg level in solid food, with crisp bread exhibiting the highest levels. In drinks, the highest value (29 ,g/L) was found in regular coffee drinks. [source] High-performance liquid chromatography with diode array detection coupled to electrospray time-of-flight and ion-trap tandem mass spectrometry to identify phenolic compounds from a Cistus ladanifer aqueous extractPHYTOCHEMICAL ANALYSIS, Issue 4 2010S. Fernández-Arroyo Abstract Introduction , Cistus ladanifer is an aromatic shrub that is widespread in the Mediterranean region. The labdanum exudate is used in the fragrance industry and has been characterised. However, there is not enough information about the phenolic content of the raw plant, the aerial part of it being a very rich source of bioactive compounds. Objective , Characterisation of the bioactive compounds of the raw plant and its aerial parts. Methodology , High-performance liquid chromatography with diode array and electrospray ionisation mass spectrometric detection was used to carry out the comprehensive characterisation of a Cistus ladanifer shrub aqueous extract. Two different MS techniques were coupled to HPLC: time-of-flight mass spectrometry and tandem mass spectrometry. Results , Many well-known compounds present in Cistus ladanifer were characterised, such as flavonoids, phenolic acids, ellagitanins, hexahydroxydiphenoyl and derivatives, and other compounds. Conclusion , The method described simultaneously separated a wide range of phenolic compounds and the proposed characterisation of the major compounds of this extract was carried out. It is important to highlight that, to our knowledge, this is the first time that a Cistus ladanifer aqueous extract from the raw plant has been characterised. Copyright © 2009 John Wiley & Sons, Ltd. [source] Gas-phase behavior of noncovalent transmembrane segment complexesRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 24 2008Linda M. M. Weigang Specific helix oligomerization between transmembrane segments (TMSs) is often promoted by motifs like GxxxG. Disruption of this motif in the transmembrane segments of vesicular stomatitis virus G-protein and of glycophorin A results in a reduced dimerization level studied by in vivo systems like ToxR. This paper reports the influence of sequence motifs like GxxxG in solution and the gas phase. The transmembrane segments may behave differently in the gas and liquid phase, because of the absence of surrounding solvent molecules in the gas phase. Comparison of experiments depending on peptide properties performed in the gas and liquid phase discloses that the peptides retain ,some memory' of their liquid-phase structure in the gas phase. A direct correlation has been found between helicity in solution as determined by circular dichroism and dimerization in the gas phase monitored by electrospray mass spectrometry. These results show that a proper folding in solution is required for oligomerization. On the other hand, sequence-specific oligomerization depending on the GxxxG motif was not observed with the mass spectrometric detection. Further on, neither concentration-dependent complex studies nor studies regarding complex stability in the gas phase , via collision-induced dissociation (CID) , led to sequence-specific differences. Finally, the findings show that in mass spectrometric measurements noncovalent interactions of studied TMSs is rather more dependent on the secondary structure and proper folding than on their primary structure. Copyright © 2008 John Wiley & Sons, Ltd. [source] Simultaneous determination of mercapturic acids derived from ethylene oxide (HEMA), propylene oxide (2-HPMA), acrolein (3-HPMA), acrylamide (AAMA) and N,N -dimethylformamide (AMCC) in human urine using liquid chromatography/tandem mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 17 2008Thomas Schettgen Mercapturic acids are highly important and specific biomarkers of exposure to carcinogenic substances in occupational and environmental medicine. We have developed and validated a reliable, specific and very sensitive method for the simultaneous determination of five mercapturic acids derived from several high-production chemicals used in industry, namely ethylene oxide, propylene oxide, acrylamide, acrolein and N,N -dimethylformamide. Analytes are enriched and cleaned up from urinary matrix by offline solid-phase extraction. The mercapturic acids are subsequently separated by means of high-performance liquid chromatography on a Luna C8 (2) column and specifically quantified by tandem mass spectrometric detection using isotopically labelled analytes as internal standards. The limits of detection (LODs) for N -acetyl- S -2-carbamoylethylcysteine (AAMA) and N -acetyl- S -2-hydroxyethylcysteine (HEMA) were 2.5,µg/L and 0.5,µg/L urine, while for N -acetyl- S -3-hydroxypropylcysteine (3-HPMA), N -acetyl- S -2-hydroxypropylcysteine (2-HPMA) and N -acetyl- S -(N -methylcarbamoyl)cysteine (AMCC) it was 5,µg/L. These LODs were sufficient to detect the background exposure of the general population. We applied the method on spot urine samples of 28 subjects of the general population with no known occupational exposure to these substances. Median levels for AAMA, HEMA, 3-HPMA, 2-HPMA and AMCC in non-smokers (n,=,14) were 52.6, 2.0, 155, 7.1 and 113.6,µg/L, respectively. In smokers (n,=,14), median levels for AAMA, HEMA, 3-HPMA, 2-HPMA and AMCC were 243, 5.3, 1681, 41.7 and 822,µg/L, respectively. Due to the simultaneous quantification of these mercapturic acids, our method is well suited for the screening of workers with multiple chemical exposures as well as the determination of the background excretion of the general population. Copyright © 2008 John Wiley & Sons, Ltd. [source] Determination of paclitaxel in human plasma following the administration of Genaxol or Genetaxyl by liquid chromatography/tandem mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 14 2006Erin R. Gardner A sensitive and specific assay for paclitaxel in plasma has been developed to overcome limitations in previously published assays, using liquid chromatography with tandem mass spectrometric detection. Plasma samples (100,µL) were subjected to liquid-liquid extraction with 1-chlorobutane/acetonitrile (4:1, v/v), with [2H5]paclitaxel employed as the internal standard. Chromatography was carried out with a Waters SymmetryShield C8 column (50,×,2.1 mm, 3.5,µm). The total run time, including equilibration, was 8 min, using a gradient of acetonitrile and 10,mM ammonium formate, pH 4.0. The assay is accurate and precise over the range of 2,2500,ng/mL and has been successfully applied to study the clinical pharmacokinetics of two formulations of paclitaxel, Genaxol and Genetaxyl, given orally and intravenously. Copyright © 2006 John Wiley & Sons, Ltd. [source] Separation of a BMS drug candidate and acyl glucuronide from seven glucuronide positional isomers in rat plasma via high-performance liquid chromatography with tandem mass spectrometric detectionRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 11 2006Y.-J. Xue A high-performance liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the determination of a BMS drug candidate and its acyl glucuronide (1- O - , glucuronide) in rat plasma. A 50-µL aliquot of each plasma sample was fortified with acetonitrile containing the internal standard to precipitate proteins and extract the analytes of interest. After mixing and centrifugation, the supernatant from each sample was transferred to a 96-well plate and injected into an LC/MS/MS system. Chromatographic separation was achieved isocratically on a Phenomenex Luna C18, 3,mm,×,150,mm, 3,µm column. The mobile phase contained 0.075% formic acid in 70:30 (v/v) acetonitrile/water. Under the optimized chromatographic conditions, the BMS drug candidate and its acyl glucuronide were separated from its seven glucuronide positional isomers within 10,min. Resolution of the parent from all glucuronides and acyl glucuronide from its positional isomers was critical to avoid their interference with quantitation of parent or acyl glucuronide. Detection was by positive ion electrospray MS/MS on a Sciex API 4000. The standard curve, which ranged from 5 to 5000,ng/mL, was fitted to a 1/x2 weighted quadratic regression model for both the BMS drug candidate and its acyl glucuronide. Whole blood and plasma stability experiments were conducted to establish the sample collection, storage, and processing conditions. The validation results demonstrated that this method was rugged and repeatable. The same methodology has also been used in mouse and human plasma for the determination of the BMS drug candidate and its acyl glucuronide. Copyright © 2006 John Wiley & Sons, Ltd. [source] High sensitivity determination of valproic acid in mouse plasma using semi-automated sample preparation and liquid chromatography with tandem mass spectrometric detectionRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 24 2005Vincenzo Pucci A high-throughput liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) assay using automated sample preparation has been developed for the determination of valproic acid (VPA) in mouse plasma. A liquid-handling system was programmed to prepare calibration standard solutions in plasma, as well as quality controls and clinical samples. Plasma protein precipitation was performed on a 96-well plate, and the collected supernatant was directly injected into a reversed-phase LC/ESI-MS/MS system in the negative ionization mode. The calibration curve for VPA was linear over a dynamic range of 0.15,100,µg/mL. The limit of detection was 75,ng/mL and the lower limit of quantitation was 150,ng/mL. Intra- and inter-day validation assays of the semi-automated plasma analysis showed satisfactory accuracy and precision. Copyright © 2005 John Wiley & Sons, Ltd. [source] A direct injection high-throughput liquid chromatography tandem mass spectrometry method for the determination of a new orally active ,v,3 antagonist in human urine and dialysateRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 22 2003Wei Zeng A generic high-throughput liquid chromatography (HTLC) tandem mass spectrometry (MS/MS) assay for the determination of compound I in human urine and dialysate (hemodialysis) was developed and validated. By using the HTLC on-line extraction technique, sample pretreatment was not necessary. The sample was directly injected onto a narrow bore large particle size extraction column (50,×,1.0,mm, 60,,m) where the sample matrix was rapidly washed away using a high flow rate (5,mL/min) aqueous mobile phase while analytes were retained. The analytes were subsequently eluted from the extraction column onto an analytical column using an organic-enriched mobile phase prior to mass spectrometric detection. The analytes were then eluted from the analytical column to the mass spectrometer for the determination. The linear dynamic range was 2.0,6000,ng/mL for the urine assay and 0.1,300,ng/mL for the dialysate assay. Intraday accuracy and precision were evaluated by analyzing five replicates of calibration standards at all concentrations used to construct the standard curve. For the urine assay, the precision (RSD%, n,=,5) ranged from 1.9 to 8.0% and the accuracy ranged from 87.8 to 105.2% of nominal value. For the dialysate assay, the precision (RSD%, n,=,5) ranged from 1.1 to 10.0% and the accuracy from 94.5 to 105.2% of nominal value. In-source fragmentation of the acyl glucuronide metabolite (compound III) did not interfere with the determination of parent compound I. The developed HTLC/MS/MS methodology was specific for compound I in the presence of compound III. Column life-time is increased and sample analysis time is decreased over traditional reversed-phase methods when direct injection assays for urine and dialysate are coupled with the technology of HTLC. Copyright © 2003 John Wiley & Sons, Ltd. [source] Metabolite profiling in rat urine by liquid chromatography/electrospray ion trap mass spectrometry.RAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 22 2003Application to the study of heavy metal toxicity This work reports the use of reverse-phase liquid chromatography coupled to electrospray ion trap (QIT) mass spectrometry for the analysis of the metabolome in rat urine. An injection of 20,,L of urine into the chromatographic system is followed by a slow gradient elution and mass spectrometric detection in the scanning mode from m/z 100,1000 in both positive and negative modes. Over a time scale of 90,min, 30 and 20 resolved peaks were observed in the positive and the negative modes, respectively, corresponding to the presence of a few hundred m/z ratios. By using a QIT analyzer, data-dependent tandem mass spectrometry of selected m/z ratios identified several compounds in rat urine and characterized various chemical families, including carboxylic acids, amines, sulfated compounds, glucuronides and glycosides, by the observation of characteristic fragment ions or neutral losses. The method has been applied to the investigation of the chronic toxicity of heavy metals in rat urine. A few tens of m/z ratios, differing in intensity more than threefold from control values, were observed in both positive and negative modes. The time variations for some selected ions suggest that LC/ESI-MS could allow selective characterization of biomarkers in response to specific toxic compounds. Copyright © 2003 John Wiley & Sons, Ltd. [source] Quantification of arecoline (areca nut alkaloid) in neonatal biological matrices by high-performance liquid chromatography/electrospray quadrupole mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 17 2003Simona Pichini A high-performance liquid chromatography (HPLC) method with mass spectrometric detection is described for determination of arecoline in newborn meconium, urine and cord serum, using pilocarpine as internal standard. The analytes were extracted from neonatal biological matrices with chloroform/isopropanol (95:5, v/v) at alkaline pH. Extracts were analyzed by HPLC coupled to an electrospray (ESI) interface and a quadrupole mass spectrometer. Chromatography was performed on a C8 reversed-phase column using 10 mM ammonium acetate (pH 4.3)/acetonitrile (90:10, v/v) as mobile phase. The mass spectrometer was operated in selected ion monitoring mode. The method was validated over the concentration range 0.005,1.00,,g/g meconium, 0.004,1.00,,g/mL cord serum and 0.001,1.00,,g/mL urine. Mean recoveries ranged between 86.5 and 90.7% for arecoline in the different biological matrices, with precision always better than 10%. The quantification limits of arecoline were 0.005,,g/g meconium, 0.004,,g/mL cord serum, and 0.001,,g/mL urine. The method was applied to the analysis of neonatal biological matrices to assess eventual fetal exposition to arecoline. Two newborns from Asian mothers who declared areca nut consumption presented arecoline in meconium with concentrations in the range 0.006,0.008,,g/g; also the urine from one neonate tested positive for the drug. Copyright © 2003 John Wiley & Sons, Ltd. [source] Selective solid-phase isolation of methionine-containing peptides and subsequent matrix-assisted laser desorption/ionisation mass spectrometric detection of methionine- and of methionine-sulfoxide-containing peptidesRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 16 2003Tom Grunert Methionine residues and the oxidised forms in proteins are becoming more and more important in view of their biological function. In particular, methionine sulfoxide seems to have a regulatory function. This paper presents a fast strategy for simultaneous determination of methionine- and methionine-sulfoxide-containing peptides, involving application of methionine-specific solid-phase reagent chemistry combined with matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS). In the first step, methionine-containing peptides are covalently bound as sulfonium salts to glass beads, whereas methionine-sulfoxide-containing peptides and other methionine-free peptides are not bound and are washed out. The wash solution is used for MALDI-MS analysis to determine the molecular masses of these peptides and to perform, if necessary, seamless post-source decay (PSD) fragment ion analysis. Methionine-sulfoxide-containing peptides can be identified due to the characteristic metastable loss of methanesulfenic acid from the protonated molecules. In the second step, the bound peptides are cleaved from the matrix of the beads by addition of 2-mercaptoethanol at pH,8.5,8.8. The resulting peptides, mainly methionine-containing peptides, are analysed in a straightforward manner by MALDI-MS and seamless PSD. The strategy allows the fast identification of methionine- and methionine-sulfoxide-containing peptides even in complex tryptic digests, as demonstrated here for the glycoprotein antithrombin. These results show that sometimes methionine-containing tryptic peptides are not detected due to steric restrictions (e.g. glycosylation near the methionine residue) on the binding reaction, and that, on the other hand, some methionine-free peptides can be quite strongly bound non-covalently to the matrix of the beads. The latter observation indicates the necessity of seamless PSD fragment ion analysis for unambiguous identification. Furthermore, there are indications that oxidation of some methionine residues occurred to a minor extent during the solid-phase isolation steps. Copyright © 2003 John Wiley & Sons, Ltd. [source] | ||||||||||||||||