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Calibration Curves (calibration + curve)
Kinds of Calibration Curves Selected AbstractsAmmonium perfluorooctanoate as a volatile surfactant for the analysis of N -methylcarbamates by MEKC-ESI-MSELECTROPHORESIS, Issue 22 2006Geert 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] Validation of an LC,MS Method for the Detection and Quantification of BZP and TFMPP and their Hydroxylated Metabolites in Human Plasma and its Application to the Pharmacokinetic Study of TFMPP in Humans,JOURNAL OF FORENSIC SCIENCES, Issue 5 2010Ushtana Antia M.Sc. Abstract:, An LC,MS method was developed for benzylpiperazine (BZP) and trifluoromethylphenylpiperazine (TFMPP), constituents of "party pills" or "legal herbal highs," and their metabolites in human blood plasma. Compounds were resolved using a mixture of ammonium formate (pH 4.5, 0.01 M) and acetonitrile (flow rate of 1.0 mL/min) with a C18 column. Calibration curves were linear from 1 to 50 ng/mL (R2 > 0.99); the lower limit of quantification (LLOQ) was 5 ng/mL; the accuracy was >90%; the intra- and interday relative standard deviations (R.S.D) were <5% and <10%, respectively. Human plasma concentrations of TFMPP were measured in blood samples taken from healthy adults (n = 6) over 24 h following a 60-mg oral dose of TFMPP: these peaked at 24.10 ng/mL (±1.8 ng/mL) (Cmax) after 90 min (Tmax). Plasma concentrations of 1-(3-trifluoromethyl-4-hydroxyphenyl) piperazine peaked at 20.2 ng/mL (±4.6 ng/mL) after 90 min. TFMPP had two disposition phases (t½ = 2.04 h (±0.19 h) and 5.95 h (±1.63 h). Apparent clearance (Cl/F) was 384 L/h (±45 L/h). [source] A Fatal Case of Suspected Anaphylaxis with Cefoperazone and Sulbactam: LC-MS AnalysisJOURNAL OF FORENSIC SCIENCES, Issue 1 2008Kenji Tsujikawa M.S. Abstract: Cefoperazone and sublactam are prescribed in combination and used in the treatment of moderate to severe bacterial infections. Serious anaphylaxis is a rare side effect. This report describes a fatal case of suspected anaphylaxis after intravenous administration of a combination of the two drugs. Heart blood was analyzed for cefoperazone by protein precipitation with acetonitrile and by liquid-liquid precipitation for sublactam after protein precipitation with aqueous acetonitrile, followed by tandem mass spectrometry in the product ion scan mode for identification and by liquid chromatography mass spectrometry in the selected ion monitoring mode for quantitation. Calibration curves for cefoperazone and sublactam were linear over the range 0.07 to 1.93 and 0.046 to 0.914 ,g/ml respectively. The decedent's blood concentrations of cefoperazone and sublactam were 0.368 and 0.143 ,g/ml respectively. As these concentrations were below concentrations reported after single dosing studies and below those considered to be minimally inhibitory, death was presumed to have been caused by hypersensitivity and not an overdose. In conclusion, this procedure is useful for detecting and quantitating cefoperazone and sublactam in postmortem blood and may be useful in the evaluation of anaphylaxis. [source] Detection and validated quantification of nine herbal phenalkylamines and methcathinone in human blood plasma by LC-MS/MS with electrospray ionizationJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 2 2007Jochen Beyer Abstract The herbal stimulants Ephedra species, Catha edulis (khat), and Lophophora williamsii (peyote) have been abused for a long time. In recent years, the herbal drug market has grown owing to publicity on the Internet. Some ingredients of these plants are also ingredients of cold remedies. The aim of the presented study is to develop a multianalyte procedure for detection and validated quantification of the phenalkylamines ephedrine, pseudoephedrine, norephedrine, norpseudoephedrine, methylephedrine, methylpseudoephedrine, cathinone, mescaline, synephrine (oxedrine), and methcathinone in plasma. After mixed-mode solid-phase extraction of 1 ml of plasma, the analytes were separated using a strong cation exchange separation column and gradient elution. They were detected using a Q-Trap LC-ESI-MS/MS system (MRM mode). Calibration curves were used for quantification using norephedrine- d3, ephedrine- d3, and mescaline- d9 as internal standards. The method was validated according to international guidelines. The assay was selective for the tested compounds. It was linear from 10 to 1000 ng/ml for all analytes. The recoveries were generally higher than 70%. Accuracy ranged from , 0.8 to 20.0%, repeatability from 2.5 to 12.3%, and intermediate precision from 4.6 to 20.0%. The lower limit of quantification was 10 ng/ml for all analytes. No instability was observed after repeated freezing and thawing or in processed samples. The applicability of the assay was tested by analysis of authentic plasma samples after ingestion of different cold medications containing ephedrine or pseudoephedrine, and after ingestion of an aqueous extract of Herba Ephedra. After ingestion of the cold medications, only the corresponding single alkaloids were detected in human plasma, whereas after ingestion of the herb extract, all six ephedrines contained in the plant were detected. The presented LC-MS/MS assay was found applicable for sensitive detection and accurate and precise quantification of all studied analytes in plasma. Copyright © 2006 John Wiley & Sons, Ltd. [source] Estimation of Individual Sennosides in Plant Materials and Marketed Formulations by an HPTLC MethodJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 4 2000SHAILESH A. SHAH Senna is a well-known drug, used in the Ayurvedic and Allopathic systems of medicine, and is a treatment for constipation. The purgative action of senna and its formulations is due to the presence of sennosides A and B. An HPTLC method has been developed for the determination of individual sennosides (A, B, C, D) without any derivatization in marketed formulations (three tablet formulations, two granule formulations and one liquid formulation) and plant materials (senna leaf and pod). The methanolic solution of a sample was applied on a pre-coated silica gel G60 F254 TLC plate (E. Merck.) and was developed using n-propanol: ethyl acetate: water: glacial acetic acid (3:3:2:0.1 v/v) as the mobile phase. The relative band speeds (Rf values) obtained were 0.35, 0.25, 0.61, 0.46 for sennosides A, B, C and D, respectively. The densitometric response was monitored at 366 nm. Calibration curves were found to be linear in the concentration ranges 193,1356, 402,2817, 71,497 and 132,927 ng per spot for sennosides A, B, C, and D, respectively. The correlation coefficients were found to be 0.9978, 0.9987, 0.9939 and 0.9983 respectively for sennosides A, B, C and D. The result obtained with the HPTLC method for total sennoside content was compared with the results using the pharmacopoeial methods (spectrophotometric (British Pharmacopoeia) and spectrofluorimetric (United States Pharmacopeia) using the ,F' test). The results revealed no significant difference in the three different methods for estimation of total sennoside. The proposed HPTLC method was found to be simple, specific, precise, accurate and rapid. It can be used for routine quality control of sennosides or senna-containing formulations for individual sennosides. [source] Comparison of HPLC and CZE methods for analysis of DOTA-like esters , reaction intermediates in synthesis of magnetic resonance contrast agentsJOURNAL OF SEPARATION SCIENCE, JSS, Issue 4-5 2010Anna Hamplová Abstract RP-HPLC and non-aqueous CZE methods were evaluated for qualitative and quantitative analysis of esterified macrocyclic compounds of the DOTA family (DOTA=1,4,7,10-tetraazacyclododecan-1,4,7,10-tetraacetic acid). This group of compounds represents important reaction intermediates in synthesis of magnetic resonance contrast agents. Calibration curves in the concentration range from 1.0×10,5 to 1.0×10,3,mol/L were plotted and the experimental data were critically compared in terms of their repeatability, linearity, LOD and LOQ. The optimized methods were successfully applied to the analysis of a real reaction mixture without any further pretreatment. [source] Quantitative determination of Aconitum alkaloids in blood and urine samples by high-performance liquid chromatographyPHYTOCHEMICAL ANALYSIS, Issue 1 2004Zhao Hong Wang Abstract An HPLC method has been developed for the simultaneous determination of the toxic Aconitum alkaloids, aconitine, mesaconitine and hypaconitine in blood and urine samples. The samples were initially subjected to solid phase extraction using Oasis® MCX cartridges, and the alkaloids were separated on an XTerraÔ RP18 column, gradient-eluted with acetonitrile: ammonium hydrogen carbonate buffer. Calibration curves were linear in the range 2.75,550 ng for aconitine and hypaconitine, and 3,600 ng for mesaconitine: the limit of detection was 0.1 ng (signal-to-noise ratio of 3) for each alkaloid. The described analysis proved to be sensitive, rapid and economical, and will be applied in the identi,cation and determination of these alkaloids in forensic and therapeutic drug monitoring. Copyright © 2004 John Wiley & Sons, Ltd. [source] Determination of cellular redox status by stable isotope dilution liquid chromatography/mass spectrometry analysis of glutathione and glutathione disulfideRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 4 2008Peijuan Zhu Oxidation of glutathione (GSH) to glutathione disulfide (GSSG) occurs during cellular oxidative stress. The redox potential of the 2GSH/GSSG couple, which is determined by the Nernst equation, provides a means to assess cellular redox status. It is difficult to accurately quantify GSH and GSSG due to the ease with which GSH is oxidized to GSSG during sample preparation. To overcome this problem, a stable isotope dilution liquid chromatography/multiple reaction monitoring mass spectrometry (LC/MRM-MS) method has been developed using 4-fluoro-7-sulfamoylbenzofurazan (ABD-F) derivatization. ABD-F derivatization of the GSH thiol group was rapid, quantitative, and occurred at room temperature. The LC/MRM-MS method, which requires no sample clean-up, was validated within the calibration ranges of 5 to 400,nmol/mL in cell lysates for GSH and 0.5 to 40,nmol/mL in cell lysates for GSSG. Calibration curves prepared by adding known concentrations of GSH and GSSG to cell lysates were parallel to the standard curve prepared in buffers. GSH and GSSG concentrations were determined in two monocyte/macrophage RAW 267.4 cell lines with or without 15-LOX-1 expression (R15LO and RMock cells, respectively) after treatment with the bifunctional electrophile 4-oxo-2(E)-nonenal (ONE). R15LO cells synthesized much higher concentrations of the lipid hydroperoxide, 15(S)-hydroperoxyeicosatetraenoic acid (15-HPETE), which undergoes homolytic decomposition to ONE. GSH was depleted by ONE treatment in both RMock and R15LO cells, leading to significant increases in their redox potentials. However, R15LO cells had higher GSH concentrations (most likely through increased GSH biosynthesis) and had increased resistance to ONE-mediated GSH depletion than RMock cells. Consequently, R15LO cells had lower reduction potentials at all concentrations of ONE. GSSG concentrations were higher in R15LO cells after ONE treatment when compared with the ONE-treated RMock cells. This suggests that increased expression of 15(S)-HPETE modulates the activity of cellular GSH reductases or the transporters involved in removal of GSSG. Copyright © 2008 John Wiley & Sons, Ltd. [source] Development of a liquid chromatography/time-of-flight mass spectrometric method for the simultaneous determination of trichothecenes, zearalenone and aflatoxins in foodstuffsRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 9 2006Hiroki Tanaka A liquid chromatography/atmospheric pressure chemical ionization mass spectrometry (LC/APCI-MS) method based on time-of-flight MS (TOFMS) with a real-time reference mass correction technique was developed for the simultaneous determination of Fusarium mycotoxins (nivalenol, deoxynivalenol, fusarenon X, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, HT-2 toxin, T-2 toxin, diacetoxyscirpenol, zearalenone) and Aspergillus mycotoxins (aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2) in corn, wheat, cornflakes and biscuits. Samples were cleaned up with a MultiSep #226 column. Detection of the mycotoxins was carried out in exact mass chromatograms with a mass window of 0.03,Th. Calibration curves were linear from 2 to 200,ng,·,mL,1 for trichothecenes and zearalenone, and 0.2 to 20,ng,·,mL,1 for aflatoxins, by 20,µL injection. The limits of detection ranged from 0.1 to 6.1,ng,·,g,1 in foodstuffs analyzed in this study. The LC/TOFMS method was found to be suitable for the screening of multiple mycotoxins in foodstuffs rapidly and with high sensitivity, and its performance was demonstrated for the confirmation for target mycotoxins. Copyright © 2006 John Wiley & Sons, Ltd. [source] Simultaneous determination of t,t -muconic, S -phenylmercapturic and S -benzylmercapturic acids in urine by a rapid and sensitive liquid chromatography/electrospray tandem mass spectrometry methodRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 17 2004Anna Barbieri We describe a rapid and sensitive high-performance liquid chromatography/electrospray tandem mass spectrometry (HPLC/ESI-MS/MS) method for simultaneous determination of the most relevant metabolites of benzene and toluene, t,t- muconic acid (t,t -MA), S -phenylmercapturic acid (S-PMA), and S -benzylmercapturic acid (S-BMA). Urine samples were purified before analysis by solid-phase microextraction (SPE) on SAX cartridges with 50,mg sorbent mass. The developed method fulfils all the standard requirements of precision and accuracy. Calibration curves were linear within the concentration range of the standards (0,80,,g/Lurine for t,t -MA, and 0,25,,g/Lurine for S-PMA and S-BMA), and had correlation coefficients ,0.997. Limits of detection were 6.0,,g/L for t,t -MA, 0.3,,g/L for S-PMA, and 0.4,,g/L for S-BMA. The method was used to determine t,t -MA, S-PMA and S-BMA levels in urine of 31 gasoline-station workers, with personal monitoring data obtained from radial symmetry passive diffusive samplers. In the context of mean work-shift exposures of 75.9,,g/m3 (range 9.4,220.2) for benzene and 331.9,,g/m3 (78.2,932.1) for toluene, metabolite concentrations in end-of-shift urine samples ranged from 23.5,275.3,,g/gcreatinine for t,t -MA, non-detectable to 0.9,,g/gcreatinine for S-PMA, and 3.8,74.8,,g/gcreatinine for S-BMA. No significant correlation was found between the environmental concentrations and urinary metabolites (p,>,0.05 for all cases); the ratios of benzene metabolites could be influenced by exposure levels and co-exposure to xylenes and toluene. The high throughput of this procedure should facilitate exploration of the metabolic effects of benzene-related co-exposure to toluene and alkylbenzenes in large populations of subjects exposed to gasoline. Copyright © 2004 John Wiley & Sons, Ltd. [source] Development of a liquid chromatographic time-of-flight mass spectrometric method for the determination of unlabelled and deuterium-labelled , -tocopherol in blood componentsRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 24 2003Wendy L. Hall A method is described for the analysis of deuterated and undeuterated , -tocopherol in blood components using liquid chromatography coupled to an orthogonal acceleration time-of-flight (TOF) mass spectrometer. Optimal ionisation conditions for undeuterated (d0) and tri- and hexadeuterated (d3 or d6) , -tocopherol standards were found with negative ion mode electrospray ionisation. Each species produced an isotopically resolved single ion of exact mass. Calibration curves of pure standards were linear in the range tested (0,1.5,,M, 0,15,pmol injected). For quantification of d0 and d6 in blood components following a standard solvent extraction, a stable-isotope-labelled internal standard (d3- , -tocopherol) was employed. To counter matrix ion suppression effects, standard response curves were generated following identical solvent extraction procedures to those of the samples. Within-day and between-day precision were determined for quantification of d0- and d6-labelled , -tocopherol in each blood component and both averaged 3,10%. Accuracy was assessed by comparison with a standard high-performance liquid chromatography (HPLC) method, achieving good correlation (r2,=,0.94), and by spiking with known concentrations of , -tocopherol (98% accuracy). Limits of detection and quantification were determined to be 5 and 50,fmol injected, respectively. The assay was used to measure the appearance and disappearance of deuterium-labelled , -tocopherol in human blood components following deuterium-labelled (d6) RRR - , -tocopheryl acetate ingestion. The new LC/TOFMS method was found to be sensitive, required small sample volumes, was reproducible and robust, and was capable of high throughput when large numbers of samples were generated. Copyright © 2003 John Wiley & Sons, Ltd. [source] Determination of , -tocopherol in infant foods by liquid chromatography combined with atmospheric pressure chemical ionisation mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 7 2003Andras Kalman A novel, sensitive and specific method for the quantification of , -tocopherol in two infant foods (milk and cereals) using liquid chromatography on-line with positive atmospheric pressure chemical ionisation mass spectrometry detection (LC/APCI-MS) has been developed. The samples were first saponified in order to eliminate fats and to transform tocopherol esters into free tocopherol, followed up by a liquid,liquid extraction of the analyte in petroleum benzine/diisopropyl ether (75:25, v/v) prior to injection onto the LC system. For the quantification, deuterium-labelled tocopherol was used as internal standard and the samples were monitored in selected ion monitoring (SIM) mode. Calibration curves between 1,40,,g/mL of , -tocopherol showed a good linear correlation (r2,=,0.99994), and the detection limit was determined to be 2.5,ng/mL. The within-day and between-day precision were determined for several dietetic infant formulae and certified reference samples, and found to be below 3.5%. The accuracy determined on a Nestlé reference sample (milk powder) was calculated to be 115.2,±,1.2%, which confirms the robustness of the proposed method. This study shows that single quadrupole LC/MS can be applied for the quantification of vitamins in food and the method offers better sensitivity and selectivity than traditional method such as LC-UV. This would simplify the preparation of the food samples and consequently enhance the vitamin analysis throughput in the food area. Copyright © 2003 John Wiley & Sons, Ltd. [source] Development and validation of a rapid and sensitive HPLC method for the quantification of 5-fluorocytosine and its metabolitesBIOMEDICAL CHROMATOGRAPHY, Issue 5 2010Kinta M. Serve Abstract To study the intracellular metabolism of the prodrug 5-fluorocytosine (5FC), we developed a novel reverse-phase high-performance liquid chromatography method to simultaneously detect 5FC and its four major anabolic metabolites: 5-fluorouracil, 5-fluorouridine, 5-fluorouridine-monophosphate and 5-fluoro-2,deoxyuridine-5,-monophosphate. Separation of each compound was accomplished under isocratic conditions using a C18 column and mobile phase of formic acid,water (1,:,99,v/v). The method was validated for both accuracy and reproducibility in cell culture media. Additionally, metabolites were assessed for stability at ambient temperatures and following freeze,thaw cycles. Calibration curves were linear over a range of 1,200,,g/mL. Limit of quantification for four of the five compounds was 1,,g/mL in cell culture media (RSD < 11%). This method was successfully used to monitor intracellular conversion of 5FC to its metabolic products over a 24h period. Copyright © 2009 John Wiley & Sons, Ltd. [source] Development and validation of UPLC-MS/MS method for simultaneous determination of gestodene and ethinyl estradiol in rat plasmaBIOMEDICAL CHROMATOGRAPHY, Issue 2 2010Zhili Xiong Abstract A selective and sensitive ultra-performance liquid chromatography method with tandem mass spectrometric detection for simultaneous determination of gestodene (GES) and ethinyl estradiol (EE) in rat plasma was developed and validated. GES, EE and the internal standard, norgestrel, were extracted with ethyl acetate, derivatized (EE only) with dansyl chloride and then back-extracted into diethyl ether-hexane (2:1, v/v). The separation was performed on an ACQUITY UPLCÔ BEH C18 column with gradient elution using mobile phase consisting of acetonitrile and water (both containing 0.1% formic acid). The detection was carried out by means of electrospray ionization (ESI) mass spectrometry in positive ion mode with multiple-reaction monitoring. Calibration curves of GES and EE were linear (r2,,,0.99) over the concentration ranges 1.59,159 and 0.196,78.4,ng/mL, respectively. The intra- and inter-day precisions were not more than 6.9 and 12.9% for GES and 10.6 and 9.0% for EE, and the accuracies were ,2.5,8.0% for GES, and ,7.2,0.19% for EE, respectively. The method herein described was superior to previous methods and was applicable to the pharmacokinetic study of GES and EE in rats. Copyright © 2009 John Wiley & Sons, Ltd. [source] High-performance liquid chromatography and LC-ESI-MS method for the identification and quantification of two biologically active isomeric coumarinolignoids cleomiscosin A and cleomiscosin B in different extracts of Cleome viscosaBIOMEDICAL CHROMATOGRAPHY, Issue 12 2008Sunil K. Chattopadhyay Abstract A rapid, sensitive and simple reverse-phase high-performance liquid chromatographic,electrospray ionization,mass spectrometry method for simultaneous determination of cleomiscosin A and cleomiscosin B has been developed and validated. The isomeric coumarinolignoids cleomiscosin A (1) and cleomiscosin B (2) were separated on a Waters symmetry C18 column with a solvent system composed of acetonitrile,methanol (1:2) and acetic acid,water (0.5 : 99.5) in a gradient elution mode. The absorption at 326 nm was chosen as the measuring wavelength in which resolution and baseline separation of compounds 1 and 2 could be obtained. The identity of the two isomeric compounds 1 and 2 in the samples were determined on a triple quadrupole mass spectrometer with ESI interface operating in the positive mode. Calibration curves were linear (r2 > 0.993) over the concentration range 20,200 µg/mL for cleomiscosin A and 10,200 µg/mL for cleomiscosin B with acceptable accuracy and precision, respectively. The intra-day and inter-day precision were 1.13 and 0.82% for cleomiscosin A and 1.78 and 1.28% for cleomiscosin B, respectively. The validated method was successfully applied for the analysis of the above two compounds in different extracts of Cleome viscosa. Copyright © 2008 John Wiley & Sons, Ltd. [source] Development of a simultaneous liquid,liquid extraction and chiral derivatization method for stereospecific GC-MS analysis of amphetamine-type stimulants in human urine using fractional factorial designBIOMEDICAL CHROMATOGRAPHY, Issue 9 2008W. R. Wan Aasim Abstract A stereospecific gas chromatography,mass spectrometry analysis method for amphetamine-type stimulants in human urine was recently developed. For maximum efficiency, liquid,liquid extraction and chiral derivatization of the analytes using (R)-(,)- , -methoxy- , -(trifluoromethyl)phenylacetyl chloride were performed simultaneously. The effects of (1) use of saturated sodium chloride in 2.0 m sodium hydroxide, (2) extraction solvent volume, (3) percentage of triethylamine, (4) derivatization reagent volume, (5) sample mixing time, (6) incubation temperature and (7) incubation time on method sensitivity and variability were assessed using a two-level, eight-run Plackett,Burman design followed by a fold-over design. The use of saturated sodium chloride solution and the derivatization reagent volume were significant factors (ANOVA, p < 0.01). The saturated sodium chloride solution decreased sensitivity whereas an increased volume of derivatization reagent increased sensitivity. Calibration curves for all analytes were linear between 5 and 500 µg/L, with correlation coefficients of >0.99. Detection limits were ,2.3 µg/L and quantitation limits ,7.7 µg/L. Reproducibility was good, with relative standard deviation values at <20%. Recovery exceeded 100% for most analytes. The experimental design enabled easy and rapid identification of significant factors using a minimal number of samples. This method has good potential for studies requiring rapid and sensitive stereospecific quantification of amphetamine-type stimulants. Copyright © 2008 John Wiley & Sons, Ltd. [source] A stereospecific high-performance liquid chromatographic assay for the determination of ketoconazole enantiomers in rat plasmaBIOMEDICAL CHROMATOGRAPHY, Issue 5 2008Dalia A. Hamdy Abstract A stereospecific high-performance liquid chromatographic assay was developed for the quantitation of ketoconazole enantiomers (KTZ) in rat plasma. After protein precipitation of 100 µL plasma using acetonitrile, a wash step was performed using hexane. The supernatant was removed and KTZ enantiomers and amiodarone, the internal standard, were extracted using liquid,liquid extraction with tert-butyl methyl ether. After transfer and evaporation of the organic layer, the residue was reconstituted in mobile phase and injected into the HPLC through a chiral column. The mobile phase consisted of hexane:ethanol:2-propanol with diethyl amine, pumped at 1.5 mL/min. All components eluted within 18 min. KTZ enantiomers were baseline resolved and peaks were symmetrical in appearance with no interferences. Calibration curves were linear over the range 62.5,5000 ng/mL of enantiomer. The intraday and interday CV% assessments were ,19 and <13%, respectively, and mean error was <4% for both enantiomers. The lower limit of quantitation was 62.5 ng/mL for each enantiomer based on 100 µL rat plasma. In rats, plasma concentrations of (+)-KTZ were higher than those of antipode after single oral doses. The assay was shown to be sensitive and appropriate for use in pharmacokinetics study of KTZ in rat. Copyright © 2008 John Wiley & Sons, Ltd. [source] Determination of faropenem in human plasma and urine by liquid chromatography,tandem mass spectrometryBIOMEDICAL CHROMATOGRAPHY, Issue 1 2008Shouhong Gao Abstract A simple, rapid and sensitive liquid chromatography/electrospray tandem mass spectrometry (LC-MS/MS) quantitative detection method, using cefalexin as internal standard, was developed for the analysis of faropenem in human plasma and urine. After precipitation of the plasma proteins with acetonitrile, the analytes were separated on a C18 reversed-phase column with 0.1% formic acid,methanol (45:55, v/v) and detected by electrospray ionization mass spectrometry in positive multiple reaction monitoring mode. Calibration curves with good linearities (r = 0.9991 for plasma sample and r = 0.9993 for urine sample) were obtained in the range 5,4000 ng/mL for faropenem. The limit of detection was 5 ng/mL. Recoveries were around 90% for the extraction from human plasma, and good precision and accuracy were achieved. This method is feasible for the evaluation of pharmacokinetic profiles of faropenem in humans, and to our knowledge, it is the first time the pharmacokinetic of faropenem has been elucidated in vivo using LC-MS/MS. Copyright © 2007 John Wiley & Sons, Ltd. [source] Enantioselective HPLC-UV method for determination of eslicarbazepine acetate (BIA 2-093) and its metabolites in human plasmaBIOMEDICAL CHROMATOGRAPHY, Issue 11 2007Gilberto Alves Abstract Eslicarbazepine acetate (BIA 2-093) is a novel central nervous system drug undergoing clinical phase III trials for epilepsy and phase II trials for bipolar disorder. A simple and reliable chiral reversed-phase HPLC-UV method was developed and validated for the simultaneous determination of eslicarbazepine acetate, oxcarbazepine, S- licarbazepine and R -licarbazepine in human plasma. The analytes and internal standard were extracted from plasma by a solid-phase extraction using Waters Oasis® HLB cartridges. Chromatographic separation was achieved by isocratic elution with water,methanol (88:12, v/v), at a flow rate of 0.7 mL/min, on a LichroCART 250-4 ChiraDex (, -cyclodextrin, 5 µm) column at 30°C. All compounds were detected at 225 nm. Calibration curves were linear over the range 0.4,8 µg/mL for eslicarbazepine acetate and oxcarbazepine, and 0.4,80 µg/mL for each licarbazepine enantiomer. The overall intra- and interday precision and accuracy did not exceed 15%. Mean relative recoveries varied from 94.00 to 102.23% and the limit of quantification of the assay was 0.4 µg/mL for all compounds. This method seems to be a useful tool for clinical research and therapeutic drug monitoring of eslicarbazepine acetate and its metabolites S- licarbazepine, R -licarbazepine and oxcarbazepine. Copyright © 2007 John Wiley & Sons, Ltd. [source] Rapid quantification of lisinopril in human plasma by liquid chromatography/tandem mass spectrometryBIOMEDICAL CHROMATOGRAPHY, Issue 4 2007Weiwei Qin Abstract An assay based on protein precipitation and liquid chromatography/tandem mass spectrometry (LC-MS/MS) has been developed and validated for the quantitative analysis of lisinopril in human plasma. After the addition of enalaprilat as internal standard (IS), plasma samples were prepared by one-step protein precipitation using perchloric acid followed by an isocratic elution with 10 mm ammonium acetate buffer (pH adjusted to 5.0 with acetic acid),methanol (70:30, v/v) on a Phenomenex Luna 5µC18 (2) column. Detection was performed on a triple-quadrupole mass spectrometer utilizing an electrospray ionization (ESI) interface operating in positive ion and selected reaction monitoring (SRM) mode with the precursor to product ion transitions m/z 406,246 for lisinopril and m/z 349,206 for enalaprilat. Calibration curves of lisinopril in human plasma were linear (r = 0.9973,0.9998) over the concentration range 2,200 ng/mL with acceptable accuracy and precision. The limit of detection and lower limit of quantification in human plasma were 1 and 2 ng/mL, respectively. The validated LC-MS/MS method has been successfully applied to a preliminary pharmacokinetic study of lisinopril in Chinese healthy male volunteers. Copyright © 2007 John Wiley & Sons, Ltd. [source] Development and validation of fixed-time method for the determination of isoxsuprine hydrochloride in commercial dosages formsDRUG TESTING AND ANALYSIS, Issue 9 2010Dr Nafisur Rahman Abstract The main aim of this work was to develop a kinetic spectrophotometric method for the quantitative analysis of isoxsuprine hydrochloride in commercial tablets. The method is based on the reaction of isoxsuprine hydrochloride (ISx) with hydroxylamine hydrochloride and ammonium cerium (IV) nitrate in sulphuric acid medium at room temperature which resulted in the formation of yellow-coloured product peaking at 380 nm. The reaction is followed spectrophotometrically by measuring the absorbance as a function of time. Fixed time method (,A = A4,A2, where A2 and A4 refer to absorbance measurements taken at 2 and 4 min, respectively) was adopted for constructing the calibration curve which was found to be linear over the concentration range of 30,80 µgmL,1 with molar absorptivity of 5.95 × 103 L mol,1 cm,1. The method has been applied successfully to the determination of isoxsuprine hydrochloride in tablets. Statistical comparison (point and interval hypothesis tests) of the results showed that there is no significant difference between the proposed method and reference method. Copyright © 2010 John Wiley & Sons, Ltd. [source] Quantitation of valproic acid in pharmaceutical preparations using dispersive liquid-liquid microextraction followed by gas chromatography-flame ionization detection without prior derivatizationDRUG TESTING AND ANALYSIS, Issue 7 2010Hamid Reza Sobhi Abstract Dispersive liquid-liquid microextraction (DLLME), coupled with gas chromatography-flame ionization detection (GC-FID), has been successfully used for the extraction and determination of valproic acid (VPA) in pharmaceutical preparations. In the developed method, an appropriate mixture of extracting and disperser solvents was rapidly injected into an aqueous sample. Having formed a cloudy solution, the mixture was centrifuged and then the extracting solvent was sedimented at the bottom of a conical test tube. The extract was then injected into a GC system directly, without any further pretreatment. Initially, microextraction efficiency factors were optimized and the optimum experimental conditions found were as follows: tetrachloroethylene (9.0 µL) as extracting solvent; acetone (1.0 mL) as disperser solvent; 5 mL acidic aqueous sample (pH 1) without salt addition. Under the selected conditions, the calibration curve showed linearity in the range of 0.1,5.0 mg/L with regression coefficient corresponding to 0.9998. The limit of detection was found to be 0.05 mg/L. Finally, the method was applied for the determination of VPA in two different pharmaceutical preparations. A reasonable intra-assay (3.9,10.8%, n = 3) and inter-assay (5.6,11.4%, n = 3) precision illustrated the good performance of the analytical procedure. The protocol proved to be rapid and cost-effective for screening purposes. Copyright © 2010 John Wiley & Sons, Ltd. [source] Fabrication of a Sensitive Cholesterol Biosensor Based on Cobalt-oxide Nanostructures Electrodeposited onto Glassy Carbon ElectrodeELECTROANALYSIS, Issue 24 2009Abdollah Salimi Abstract Electrodeposited cobalt oxide (CoOx) nanomaterials are not only used for immobilization of cholesterol oxidase (ChOx) but also as electron transfer mediator for oxidation of H2O2 generated in the enzymatic reaction. Voltammetry and flow injection analysis (FIA) were used for determination of cholesterol. FIA determination of cholesterol with biosensors yielded a calibration curve with the following characteristics: linear range up to 50,,M, sensitivity of 43.5,nA ,M,1 cm,2 and detection limit of 4.2,,M. The apparent Michaelis-Menten constant and the response time of the biosensor are 0.49,mM and 15,s, respectively. This biosensor also exhibits good stability, reproducibility and long life time. [source] Voltammetric Detection of Lead(II) Using Amide-Cyclam- Functionalized Silica-Modified Carbon Paste ElectrodesELECTROANALYSIS, Issue 15 2009Stéphanie Goubert-Renaudin Abstract 2-(4,8,11-Triscarbamoylmethyl-1,4,8,11-tetraazacyclotetradec-1-yl)acetamide (TETAM) derivatives bearing 1, 2, or 4 silylated arms have been synthesized and grafted to the surface of silica gel and ordered mesoporous silica samples. The resulting organic-inorganic hybrids have been incorporated into carbon paste electrodes and applied to the preconcentration electroanalysis of Pb(II). The attractive recognition properties of these cyclam derivatives functionalized with amide pendent groups toward Pb(II) species and the highly porous structure of the adsorbents can be exploited for the selective and sensitive detection of the target analyte. Various parameters affecting the preconcentration and detection steps have been discussed with respect to the composition and pH of both accumulation and detection media, the nature of the adsorbent (number of silylated groups linking the macrocycle to silica, texture of materials), the accumulation time, and the presence of interfering cations. Under optimal conditions and for 2,min accumulation at open-circuit, the voltammetric response increased linearly with the Pb(II) concentration in a range extending from 2×10,7 to 10,5,M, while a longer accumulation time of 15,min afforded a linear calibration curve between 10,8 and 10,7,M with a detection limit of 2.7×10,9,M which is well below the European regulatory limit of lead in consumption water. [source] Construction of an antibody microarray based on agarose-coated slidesELECTROPHORESIS, Issue 3 2007Lin-Li Lv Abstract The antibody microarray, a high-throughput multiplex immunoassay method, has become a significant tool for quantitative proteomics studies. We describe here the strategies for optimizing the condition of antibody microarray building based on agarose-coated slides. In this study, modified glass slides were robotically printed with capture antibodies against monocyte chemoattractant protein 1 (MCP-1), then dilutions of the cytokine were applied to the arrays, and the protein was detected with biotin-labeled antibody coupled with Cy3-conjugated streptavidin. Thus a protein profiling microarray based on sandwich immunoassay has been established. Various factors in the production of antibody microarrays were analyzed: the capture antibody concentrations, shelf life of the postprinting slides, blocking buffers, and reproducibility of the system. A calibration curve with a correlation coefficient of 0.9995 was established which suggested that the matrix can retain arrayed proteins in near-quantitative fashion. The results revealed high signal uniformity and reproducibility with regard to intra-array (1.3%) and the interarray (8.7%) variation at the capture antibody concentration of 125,µg/mL. Besides, the printed arrays could be stored for at least two months without any apparent change of the performance parameters. [source] Cytochrome b559 content in isolated photosystem II reaction center preparationsFEBS JOURNAL, Issue 10 2003Inmaculada Yruela The cytochrome b559 content was examined in five types of isolated photosystem II D1-D2-cytochrome b559 reaction center preparations containing either five or six chlorophylls per reaction center. The reaction center complexes were obtained following isolation procedures that differed in chromatographic column material, washing buffer composition and detergent concentration. Two different types of cytochrome b559 assays were performed. The absolute heme content in each preparation was obtained using the oxidized-minus-reduced difference extinction coefficient of cytochrome b559 at 559 nm. The relative amount of D1 and cytochrome b559,-subunit polypeptide was also calculated for each preparation from immunoblots obtained using antibodies raised against the two polypeptides. The results indicate that the cytochrome b559 heme content in photosystem II reaction center complexes can vary with the isolation procedure, but the variation of the cytochrome b559,-subunit/D1 polypeptide ratio was even greater. This variation was not found in the PSII-enriched membrane fragments used as the RC-isolation starting material, as different batches of membranes obtained from spinach harvested at different seasons of the year or those from sugar beets grown in a chamber under controlled environmental conditions lack variation in their ,-subunit/D1 polypeptide ratio. A precise determination of the ratio using an RC1-control sample calibration curve gave a ratio of 1.25 cytochrome b559,-subunit per 1.0 D1 polypeptide in photosystem II membranes. We conclude that the variations found in the reaction center preparations were due to the different procedures used to isolate and purify the different reaction center complexes. [source] Installation age of limestone masonry determined from its viscous remagnetizationGEOARCHAEOLOGY: AN INTERNATIONAL JOURNAL, Issue 1 2006Graham John Borradaile Many rocks passively acquire some time-dependent or "viscous" remanent magnetism (VRM) at ambient temperatures, without any extraordinary energetic intervention. This magnetization overprints existing remanent magnetization so that it is effectively a remagnetization subparallel to the contemporary geomagnetic field, averaging the geomagnetic field orientation. Certain limestone masonry remagnetizes viscously over an archaeologically useful interval (100 to 8000 Ka) so that the degree of remagnetization is monotonically (but not linearly) related to the construction age. The laboratory unblocking temperature (TUB) that removes the viscous magnetization is a simple monotonic measure of relative age. The longer a piece of masonry remained stabilized in a certain orientation, the greater is its viscous remagnetization and the higher is its TUB. Monuments of known age with a similar limestone source permit us to establish a calibration curve of T UB against historical ages. The resulting calibration curve may then be used to predict the ages of otherwise-undated masonry. Viscous remanent magnetism dating provides precision of <50a in medieval monuments in England and <150a precision for classical to Neolithic monuments in Cyprus; precision depends on the remagnetization rate of the limestone in question. Our calibration curves, for the Jurassic Oolitic Limestone of England and for the Lefkara-Pakhna Chalks of Cyprus, allowed us to investigate the authenticity of a medieval English synagogue in Lincoln, England, and of a medieval house in Cyprus. Multiple archaeologic VRMs show that masonry was recycled in historical times. © 2006 Wiley Periodicals, Inc. [source] Using quantitative real-time PCR to detect salmonid prey in scats of grey Halichoerus grypus and harbour Phoca vitulina seals in Scotland , an experimental and field studyJOURNAL OF APPLIED ECOLOGY, Issue 2 2008I. Matejusová Summary 1There is considerable debate over the impact of seal predation on salmonid populations in both the Atlantic and Pacific oceans. Conventional hard-part analysis of scats has suggested that salmonids represent a minor component of the diet of grey seals (Halichoerus grypus) and harbour seals (Phoca vitulina) in the UK. However, it is unclear whether this is an accurate reflection of the diet or due to methodological problems. To investigate this issue, we applied quantitative PCR (qPCR) to examine the presence of salmonids in the diet of seals in the Moray Firth, UK, during the summers of 2003 and 2005. 2Two qPCR assays were designed to detect Atlantic salmon Salmo salar and sea trout Salmo trutta DNA in field samples and experimentally spiked scats. The proportion of scats sampled in the field that were positive for salmonid DNA was low (ª10%). However, the DNA technique consistently resulted in more positive scats than when hard-part analysis was used. 3An experimental study using spiked scat material revealed a highly significant negative relationship between Ct values obtained from the Atlantic salmon qPCR assay and the proportion of Atlantic salmon material added to scats. The Ct value denotes the cycle number at which the increasing fluorescence signal of target DNA crosses a threshold value. Ct values from field-collected seal scats suggested they contained a very low concentration of salmonid remains (1,5%) based on an approximate calibration curve constructed from the experimental data. 4Synthesis and applications. The qPCR assay approach was shown to be highly efficient and consistent in detection of salmonids from seal scats, and to be more sensitive than conventional hard-parts analysis. Nevertheless, our results confirm previous studies indicating that salmonids are not common prey for seals in these Scottish estuaries. These studies support current management practice, which focuses on control of the small number of seals that move into key salmonid rivers, rather than targeting the larger groups of animals that haul-out in nearby estuaries. [source] Raman spectroscopy for spinline crystallinity measurements.JOURNAL OF APPLIED POLYMER SCIENCE, Issue 5 2008Abstract Online Raman spectra, obtained at different points along the spinline during the melt spinning of polypropylene homopolymer (hPP) fibers, are presented. The percentage crystallinity corresponding to each spectrum was determined from the normalized intensity of the 809-cm,1 Raman band. A calibration curve for propylene crystallinity was established offline with compression-molded films and fibers spun under different processing conditions. Several hPPs and propylene,ethylene copolymers (with 5,15% ethylene) were used to cover a wide calibration range for propylene crystallinity (9.5,60.9%) with an R2 value of 0.989. This calibration curve was subsequently used to predict the polypropylene crystallinity in the spinline as a function of distance from the spinneret. Under identical conditions of quench and throughput, at a fixed point along the spinline, the overall crystallinity developed in the fiber was found to increase with an increase in the spinning speed. As the spinning speed increased, the point of the onset of crystallization moved closer to the spinneret. The rise in crystallinity was more gradual, at 750 m/min as opposed to 1500 m/min. Increasing the throughput at constant spinning speed was found to decrease the rate of crystallization because of a decrease in the spinline stress. At a fixed distance from the spinneret under identical conditions of quench and spinning speed, fibers spun at a higher throughput showed less overall crystallinity. The onset and rate of crystallization was found to be faster in the lower melt index H502-25RG resin as compared to the 5D49 resin under the spinning conditions explored. The experimental data presented here were used to validate fundamental fiber-spinning models (see part II of this series of articles). The validated models and experimental observations can be used to guide the fiber spinning of isotactic polypropylene for rapid product development. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008 [source] ORGANIC ACIDS PROFILE IN TOMATO JUICE BY HPLC WITH UV DETECTIONJOURNAL OF FOOD QUALITY, Issue 1 2007OMBRETTA MARCONI ABSTRACT A simple method was developed to determine 10 organic acids simultaneously in tomato products using reverse-phase high performance liquid chromatography (HPLC) column with the diode array detector set at 210 nm. After centrifugation and filtration, the samples were passed through to an anion exchange resin and the organic acids were released using 0.1 N HCl. The chromatographic separation was achieved with isocratic analysis in a 20-min run. The method was reliable and sensitive. The coefficient of determination of the standard calibration curve is 0.9925 , r2 , 0.9999 and the limit of detection ranged from 0.08 to 6.00 mg/kg for trans -aconitic acid and acetic acid, respectively. The limit of quantification ranged from 0.19 to 15.18 mg/kg for trans-aconitic and acetic acid, respectively. To establish the efficiency of the anion resin the procedure was applied to a standard solution of a mixture of organic acids. The organic acids recovery ranged from 87.0% ± 1.9 for citramalic acid to 109.9% ± 5.2 for fumaric acid. [source] | |||||||||||||||||||||||||||||||||||||||||||