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Analytical Column (analytical + column)
Kinds of Analytical Column Selected AbstractsSimple and simultaneous determination of sulphapyridine and acetylsulphapyridine in human serum by column-switching high-performance liquid chromatographyJOURNAL OF CLINICAL PHARMACY & THERAPEUTICS, Issue 6 2002D. Teshima PhD Summary Objective:, A high-performance liquid chromatography (HPLC) with an automated on-line column-switching system was used for the simultaneous determination of sulphapyridine and acetylsulphapyridine, two major active metabolites related to the adverse effects of sulphasalazine, in human serum. Methods:, Serum samples were directly injected into the HPLC, with the valve automatically switched on to remove serum proteins and other hydrophilic components remaining in the pre-column after elution of sulphapyridine and acetylsulphapyridine to the analytical column. Results:, Serum proteins did not interfere with the analysis of either compound. The recoveries of SLP and Ac-SLP from drug-free human serum were 93·03,99·18% and CV were 2·88,4·34%. The within-run reproducibility of assays was excellent with relative standard deviations (RSD) of 1·01,3·90% (SLP) and 0·77,5·56% (Ac-SLP). The limit of quantification of sulphapyridine and acetylsulphapyridine was 3·13 ,g/mL and 0·50 ,g/mL, respectively. The serum concentrations in a patient with ulcerative colitis, who took 1·0 g sulphasalazine twice daily, were 31·20 ,g/mL for sulphapyridine and 14·64 ,g/mL for acetylsulphapyridine at 7 h after ingestion. Conclusion:, The present simple and reproducible assay was useful for the monitoring of serum sulphapyridine and acetylsulphapyridine. [source] Ticlopidine quantification in human plasma by high-performance liquid chromatography coupled to electrospray tandem mass spectrometry.JOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 12 2004Application to bioequivalence study Abstract A rapid, sensitive and specific method to quantify ticlopidine in human plasma using clopidogrel as the internal standard (IS) is described. The analyte and the IS were extracted from acidified plasma by liquid,liquid extraction using diethyl ether,hexane (80 : 20, v/v). The extracts were analyzed by high-performance liquid chromatography coupled to electrospray tandem mass spectrometry (HPLC/MS/MS). Chromatography was performed isocratically on a Jones Genesis C8 4 µm analytical column (150 × 4.1 mm i.d.). The method had a chromatographic run time of 3.0 min and a linear calibration curve over the range 1.0,1000 ng ml,1 (r2 > 0.999427). The limit of quantification was 1.0 ng ml,1. This HPLC/MS/MS procedure was used to assess the bioequivalence of two ticlopidine 250 mg tablet formulations (ticlopidine test formulation from Apotex do Brasil, Brazil, and Ticlid from Sanofi-Synthelabo, standard reference formulation). A single 250 mg dose of each formulation was administered to healthy volunteers. The study was conducted using an open, randomized, two-period crossover design with a 2 week washout interval. Since the 90% confidence interval for Cmax and area under the curve ratios were all inside the 80,125% interval proposed by the US Food and Drug Administration, it was concluded that ticlopidine formulation from Apotex do Brasil is bioequivalent to Ticlid formulation with respect to both the rate and the extent of absorption. Copyright © 2004 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] Determination of the active metabolites of sibutramine in rat serum using column-switching HPLCJOURNAL OF SEPARATION SCIENCE, JSS, Issue 15 2008So Young Um Abstract A simple and direct analysis using column-switching HPLC method was developed and validated for the quantification of active metabolites of sibutramine, N -mono-desmethyl metabolite (metabolite 1, M1) and N -di-desmethyl metabolite (metabolite 2, M2) in the serum of rats administered sibutramine HCl (5.0 mg/kg, p.o.). Rat serum was directly injected onto the precolumn without sample prepreparation step following dilution with mobile phase A, i. e., methanol,ACN,20 mM ammonium phosphate buffer (pH 6.0 with phosphoric acid) (8.3:4.5:87.2 by volume). After the endogenous serum components were eluted to waste, the system was switched and the analytes were eluted to the trap column. Active metabolites M1 and M2 were then back-flushed to the analytical column for separation with mobile phase B, i. e., methanol,ACN,20 mM ammonium phosphate buffer (pH 6.0 with phosphoric acid) (35.8:19.2:45 by volume) and detected at 223 nm. The calibration curves of active metabolites M1 and M2 were linear in the range of 0.1,1.0 ,g/mL and 0.15,1.8 ,g/mL. This method was fully validated and shown to be specific, accurate (10.4,10.7% error), and precise (1.97,8.79% CV). This simple and rapid analytical method using column-switching appears to be useful for the pharmacokinetic study of active metabolites (M1 and M2) of sibutramine. [source] Solid phase microextraction-high performance liquid chromatographic determination of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in the presence of sodium dodecyl sulfate surfactantJOURNAL OF SEPARATION SCIENCE, JSS, Issue 12 2008Gaurav Abstract A simple and sensitive method has been developed using preconcentration technique solid phase microextraction (SPME) and analytical technique HPLC-UV for the determination of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) from the environmental samples. Aqueous solution of anionic surfactant SDS was used for the extraction of both nitramine high explosives, viz., HMX and RDX from soil samples which were subsequently sorbed on SPME fiber. The static desorption was carried out in the desorption chamber of the SPME-HPLC interface in the presence of mobile phase ACN/methanol/water (30:35:35) and the subsequent chromatographic analysis at a flow rate of 0.5 mL/min and detection at 230 nm. For this purpose, a C18, 5 ,m RP analytical column was used as a separation medium in this method. Several parameters relating to SPME, e.g., adsorption/desorption time, concentration of salt, stirring rate, etc., were optimized. The method was linear over the range of 20,400 ng/mL for HMX and RDX standards in the presence of surfactant in aqueous phase, respectively. The correlation coefficient (R2) for HMX and RDX are 0.9998 and 0.9982, respectively. With SPME, the detection limits (S/N = 3) in ng/mL are 0.05 and 0.1 for HMX and RDX, respectively in the presence of the SDS surfactant. The developed method has been applied successfully to the analysis of real environmental samples like bore well water, river water, and ground alluvial soil. [source] Simultaneous determination of diethylene glycol and propylene glycol in pharmaceutical products by HPLC after precolumn derivatization with p -toluenesulfonyl isocyanateJOURNAL OF SEPARATION SCIENCE, JSS, Issue 16 2007Tao Zhou Abstract A simple and reliable HPLC method was developed for the simultaneous quantitative analysis of diethylene glycol (DEG) and propylene glycol (PG) in pharmaceutical products by precolumn derivatization. The derivatization reagent p -toluenesulfonyl isocyanate (TSIC, 10 ,L, 20% in ACN v/v) was added to 100 ,L of the sample, and then 10 ,L of water was added. The resulting derivatives were separated using a C18 analytical column and a mobile phase composed of 0.01 M KH2PO4 buffer (adjusted to pH 2.5 with phosphoric acid) and ACN (47:53 v/v) at 1 mL/min and 25°C. For detection, UV light at 227 nm was used. The derivatization conditions including reaction time, temperature, and concentration of TSIC were optimized. The calibration curves were linear from 0.062 to 18.6 ,g/mL (r2 = 0.9999) and from 0.071 to 21.3 ,g/mL (r2 = 0.9999) for DEG and PG, respectively. The RSD values of intra- and interday assays were all below 4% for DEG and PG. The proposed method was then successfully applied to analyze two Armillarisin A injection samples and two spiked syrup samples. [source] Multiresidue HPLC analysis of ten quinolones in milk after solid phase extraction: Validation according to the European Union Decision 2002/657/ECJOURNAL OF SEPARATION SCIENCE, JSS, Issue 15 2007Eleni A. Christodoulou Abstract A rapid and sensitive analytical method was developed for the residue analysis of ten quinolones (enoxacin (ENO), ofloxacin (OFL), norfloxacin (NOR), ciprofloxacin (CIP), danofloxacin (DAN), enrofloxacin (ENR), sarafloxacin (SAR), oxolinic acid (OXO), nalidixic acid (NAL), and flumequine (FLU)) in cow's milk. The analytes were extracted from milk by a deproteinization step followed by a simple SPE cleanup procedure using LiChrolut RP-18 Merck cartridges. Recoveries varied between 75 and 92%. HPLC separation was performed at 25°C using an ODS-3 PerfectSil® Target (250×4 mm2) 5 ,m analytical column (MZ-Analysentechnik, Germany). The mobile phase consisted of a mixture of TFA 0.1%,CH3CN,CH3OH, delivered by a gradient program at the flow rate of 1.2 mL/min. Elution of the ten analytes and the internal standard (caffeine, 7.5 ng/,L) was completed within 27 min. Column effluent was monitored using a photodiode array detector, set at 275 and 255 nm. The developed method was validated according to the criteria of Commission Decision 2002/657/EC. The LODs of the specific method of quinolones' determination in milk varied between 1.5 and 6.8 ng/,L. [source] Development and validation of an HPLC confirmatory method for the determination of seven tetracycline antibiotics residues in milk according to the European Union Decision 2002/657/ECJOURNAL OF SEPARATION SCIENCE, JSS, Issue 15 2007Victoria F. Samanidou Abstract An HPLC method with diode-array detection, at 355 nm, was developed and validated for the determination of seven tetracyclines (TCs) in milk: minocycline (MNC), TC, oxytetracycline (OTC), methacycline (MTC), demeclocycline (DMC), chlortetracycline (CTC), and doxycycline (DC). Oxalate buffer (pH 4) was used with 20% TCA as a deproteinization agent for the extraction of analytes from milk followed by SPE. The separation was achieved on an Inertsil ODS-3, 5 ,m, 250×4 mm2 analytical column at ambient temperature. The mobile phase, a mixture of A: 0.01 M oxalic acid and B: CH3CN, was delivered using a gradient program. The procedure was validated according to the European Union decision 2002/657/EC determining selectivity, stability, decision limit, detection capability, accuracy, and precision. Mean recoveries of TCs from spiked milk samples (50, 100, and 200 ng/g) were 93.8,100.9% for MNC, 96.8,103.7% for OTC, 96.3,101.8% for TC, 99.4,107.2% for DMC, 99.4,102.9% for CTC, 96.3,102.7% for MTC, and 94.6,102.1% for DC. All RSD values were lower than 8.5%. The decision limits CCa calculated by spiking 20 blank milk samples at MRL (100 ,g/kg) ranged from 101.25 to 105.84 ,g/kg, while detection capability CCbfrom 103.94 to 108.88 ,g/kg. [source] Quality control of Pulsatilla koreana based on the simultaneous determination of triterpenoidal saponins by HPLC-ELSD and principal component analysisPHYTOCHEMICAL ANALYSIS, Issue 4 2010Ki Yong Lee Abstract Introduction , Pulsatilla koreana Nakai, with triterpenoidal saponins as its main pharmacological effective compounds, is known to have several biological activities, including hypoglycaemic, antitumour, cognition-enhancing, neuroprotective, cytotoxic and antiangiogenic activities. However, few analytical methods have been reported on the quality assessment of P. koreana roots. Obejective , To establish a high-performance liquid chromatography coupled with evaporative light scattering detection for the simultaneous determination of five triterpenoidal saponins, including pulsatilloside E (1), pulsatilla saponin H (2), anemoside B4 (3), hederacolchiside E (4) and cussosaponin C (5) in P. koreana. Methodology , The chromatographic separation was performed on a Shiseido CapCell PAK C18 analytical column efficiently using gradient elution with acetonitrile and water. Results , All calibration curves showed excellent linear regressions (R2 > 0.9996) within the range of tested concentrations. The intra- and inter-day variations were below 4.78% in terms of RSD. The recoveries were 94.82,102.97% with RSD of 0.27,3.92% for spiked P. koreana samples. Conclusion , The validated method was successfully used for the analysis of five saponins in P. koreana from different locations. Moreover, the different samples were clustered in accordance with contents of triterpenoidal saponins based on aglycon type by a principal component analysis. Copyright © 2010 John Wiley & Sons, Ltd. [source] Automation of nanoflow liquid chromatography-tandem mass spectrometry for proteome analysis by using a strong cation exchange trap columnPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 4 2007Xiaogang Jiang Abstract An approach was developed to automate sample introduction for nanoflow LC-MS/MS (,LC-MS/MS) analysis using a strong cation exchange (SCX) trap column. The system consisted of a 100,,m id×2,cm SCX trap column and a 75,,m id×12,cm C18 RP analytical column. During the sample loading step, the flow passing through the SCX trap column was directed to waste for loading a large volume of sample at high flow rate. Then the peptides bound on the SCX trap column were eluted onto the RP analytical column by a high salt buffer followed by RP chromatographic separation of the peptides at nanoliter flow rate. It was observed that higher performance of separation could be achieved with the system using SCX trap column than with the system using C18 trap column. The high proteomic coverage using this approach was demonstrated in the analysis of tryptic digest of BSA and yeast cell lysate. In addition, this system was also applied to two-dimensional separation of tryptic digest of human hepatocellular carcinoma cell line SMMC-7721 for large scale proteome analysis. This system was fully automated and required minimum changes on current ,LC-MS/MS system. This system represented a promising platform for routine proteome analysis. [source] Rapid screening and confirmation of drugs and toxic compounds in biological specimens using liquid chromatography/ion trap tandem mass spectrometry and automated library searchRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 1 2010Hsiu-Chuan Liu Recent advances in liquid chromatography/tandem mass spectrometry (LC/MS/MS) technology have provided an opportunity for the development of more specific approaches to achieve the ,screen' and ,confirmation' goals in a single analytical step. For this purpose, this study adapts the electrospray ionization ion trap LC/MS/MS instrumentation (LC/ESI-MS/MS) for the screening and confirmation of over 800 drugs and toxic compounds in biological specimens. Liquid-liquid and solid-phase extraction protocols were coupled to LC/ESI-MS/MS using a 1.8-µm particle size analytical column operated at 50°C. Gradient elution of the analytes was conducted using a solvent system composed of methanol and water containing 0.1% formic acid. Positive-ion ESI-MS/MS spectra and retention times for each of the 800 drugs and toxic compounds were first established using 1,10,µg/mL standard solutions. This spectra and retention time information was then transferred to the library and searched by the identification algorithm for the confirmation of compounds found in test specimens , based on retention time matches and scores of fit, reverse fit, and purity resulting from the searching process. The established method was found highly effective when applied to the analyses of postmortem specimens (blood, urine, and hair) and external proficiency test samples provided by the College of American Pathology (CAP). The development of this approach has significantly improved the efficiency of our routine laboratory operation that was based on a two-step (immunoassay and GC/MS) approach in the past. Copyright © 2009 John Wiley & Sons, Ltd. [source] A liquid chromatography/tandem mass spectrometric approach for the determination of gangliosides GD3 and GM3 in bovine milk and infant formulaeRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 24 2006Lambert K. Sřrensen A liquid chromatographic/tandem mass spectrometric method using pneumatically assisted electrospray ionisation (LC/ESI-MS/MS) was developed for the determination of gangliosides GD3 and GM3 in milk and infant formulae. The gangliosides were extracted in a chloroform/methanol/water environment and cleaned up by solid-phase extraction (SPE) on an end-capped C8 sorbent. The gangliosides were detected in negative ion mode after separation on a reversed-phase (RP) C5 analytical column. From the different ganglioside molecular species, product ions at m/z 290 corresponding to an N-acetylneuraminic acid fragment were produced in the collision cell and used in selected reaction monitoring. A standard addition technique was applied for quantification. The relative repeatability standard deviations were less than 5% for GD3 (level 10,mg/L) and 14% for GM3 (level 0.1,0.2,mg/L). Copyright © 2006 John Wiley & Sons, Ltd. [source] Increased productivity in quantitative bioanalysis using a monolithic column coupled with high-flow direct-injection liquid chromatography/tandem mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 11 2006Mike-Qingtao Huang The feasibility of using a monolithic column as the analytical column in conjunction with high-flow direct-injection liquid chromatography/tandem mass spectrometry (LC/MS/MS) to increase productivity for quantitative bioanalysis has been investigated using plasma samples containing a drug and its epimer metabolite. Since the chosen drug and its epimer metabolite have the same selected reaction monitoring (SRM) transitions, chromatographic baseline separation of these two compounds was required. The results obtained from this monolithic column system were directly compared with the results obtained from a previously validated assay using a conventional C18 column as the analytical column. Both systems have the same sample preparation, mobile phases and MS conditions. The eluting flow rate for the monolithic column system was 3.2,mL/min (with 4:1 splitting) and for the C18 column system was 1.2,mL/min (with 3:1 splitting). The monolithic column system had a run time of 5,min and the conventional C18 column system had a run time of 10,min. The methods on the two systems were found to be equivalent in terms of accuracy, precision, sensitivity and chromatographic separation. Without sacrificing the chromatographic separation, sensitivity, accuracy and precision of the method, the reduced run time of the monolithic column method increased the sample throughput by a factor of two. Copyright © 2006 John Wiley & Sons, Ltd. [source] The use of turbulent flow chromatography and the isocratic focusing effect to achieve on-line cleanup and concentration of neat biological samples for low-level metabolite analysisRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 5 2005J. L. Herman The use of turbulent flow chromatography in conjunction with column switching isocratic focusing was used to perform on-line sample cleanup and concentration of neat rat plasma for the identification of low-level metabolites. The concentration was achieved by focusing multiple injections, which were cleaned by a turbulent flow column, onto an analytical column prior to elution into the mass spectrometer. In addition, the first application of turbulent flow chromatography for on-line sample cleanup of neat bile samples is reported. The on-line cleanup and concentration method extracts and concentrates a sample 20-fold in 1,h, and is completely automated. 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] Congener-specific analysis of hexabromocyclododecane by high-performance liquid chromatography/electrospray tandem mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 13 2003Wesley Budakowski A congener-specific method based on high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ES-MS/MS) in the negative ion mode was developed for the analysis of hexabromocyclododecane (HBCDD). On a C18 analytical column, with a methanol/water mobile phase, the , -isomer was completely resolved from the , - and , -isomers while the , - and , -isomers were sufficiently resolved at half their peak heights. The ES spray voltage strongly influenced the intensity of the ion signal. For MS, a source temperature of 500°C and a collision energy of 50,eV were found to be optimum for the [M,H], to Br, transition. Run-to-run and day-to-day (n,=,3) variability was minimal, with relative standard deviations of 2.6,4.1 and 2.4,4.4%, respectively. The limit of detection was 4,6,pg on-column. When applied to tissue samples from Lake Winnipeg fish both , - and , -isomers of HBCDD were found in low-ng/g (lipid corrected) concentrations. Copyright © 2003 John Wiley & Sons, Ltd. [source] Direct-injection high performance liquid chromatography ion trap mass spectrometry for the quantitative determination of olanzapine, clozapine and N -desmethylclozapine in human plasmaRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 13 2002Manfred Kollroser A specific and sensitive direct-injection high performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS) method has been developed for the rapid identification and quantitative determination of olanzapine, clozapine, and N -desmethylclozapine in human plasma. After the addition of the internal standard dibenzepin and dilution with 0.1% formic acid, plasma samples were injected into the LC/MS/MS system. Proteins and other large biomolecules were removed during an online sample cleanup using an extraction column (1,×,50,mm i.d., 30,µm) with a 100% aqueous mobile phase at a flow rate of 4,mL/min. The extraction column was subsequently brought inline with the analytical column by automatic valve switching. Analytes were separated on a 5,µm Symmetry C18 (Waters) analytical column (3.0,×,150,mm) with a mobile phase of acetonitrile/0.1% formic acid (20:80, v/v) at a flow rate of 0.5,mL/min. The total analysis time was 6,min per sample. The inter- and intra-assay coefficients of variation for all compounds were <11%. By eliminating the need for extensive sample preparation, the proposed method offers very large savings in total analysis time. Copyright © 2002 John Wiley & Sons, Ltd. [source] Application to routine analysis of a method to determine multiclass pesticide residues in fresh vegetables by gas chromatography/tandem mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 11 2002J. L. Martínez Vidal The use of gas chromatography/tandem mass spectrometry (GC/MS/MS) applied to determine multiple pesticide residues in fresh vegetables has been thoroughly studied. A single injection method to detect, confirm and quantify 54 multiclass pesticides has been developed and applied in a routine analysis laboratory. The proposed method consists of a rapid extraction of 15,g of vegetable sample with dichloromethane. An additional clean-up step is not necessary even when injecting 10,µL of extract. Instead the gas chromatograph was fitted with a carbofrit inserted into the glass liner and a guard column. In addition, the detection mode chosen (MS/MS) provides additional selectivity. The method has been validated and applied to 1300 samples in a routine laboratory following specified quality criteria. The recovery efficiencies obtained for all the pesticides ranged between 70.2 and 110.8% at two different fortification levels. The relative standard deviation for quantification (RSD) was lower than 16.7% for all the compounds. Important experimental parameters, such as the conditioning of carbofrit, overload of the analytical column, and cleanliness of the ion trap, were evaluated for their influence on the performance of the method. Copyright © 2002 John Wiley & Sons, Ltd. [source] Determination of carboplatin in canine plasma by high-performance liquid chromatographyBIOMEDICAL CHROMATOGRAPHY, Issue 8 2010Nicolas Villarino Abstract Carboplatin is an antineoplastic drug administered to treat different tumoral conditions in canine oncology. The objective of this study was to validate a high-performance chromatographic (HPLC) method which could be applied in canine pharmacokinetic studies. Following ultrafiltration using a Centrifree device, standards, quality controls and plasma samples were separated by isocratic reversed-phase HPLC on an Inertsil ODS-2 (250 × 4.6,mm i.d.) analytical column and quantified using UV detection at 220,nm. The mobile phase was potassium phosphate (pH 4.5), with a flow-rate of 1.0,mL/min. The procedure produced a linear curve (r2 > 0.999) over the concentration range 1,200,,g/mL. The lower limit of quantification was 1,,g/mL. The intra-assay and inter-assay precision was ,90%. The overall recovery was ,90%. The method was illustrated with a preliminary pharmacokinetic analysis on nine dogs treated with carboplatin at our hospital. Carboplatin disposition followed a monocompartmental structure in dogs and was characterized by a short half-life (50,min). Copyright © 2009 John Wiley & Sons, Ltd. [source] Development of a method to measure methadone enantiomers and its metabolites without enantiomer standard compounds for the plasma of methadone maintenance patientsBIOMEDICAL CHROMATOGRAPHY, Issue 7 2010Sheng-Chang Wang Abstract A liquid chromatography,photodiode array (LC-PDA) method using a chiral analytical column was developed to determine the plasma levels of enantiomers of methadone and its chiral metabolite, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), without the standard compounds of R -form or S -form enantiomers. This method was established by the characteristics of recombinant cytochrome P-450 (CYP) isozymes, where CYP2C19 prefers to metabolize R -methadone and CYP2B6 prefers to metabolize S -methadone. We incubated the racemic methadone standard with either enzyme for 24,h. We identified the retention times of R - and S -methadone to be around 10.72 and 14.46,min, respectively. Furthermore, we determined the retention times of R - and S -EDDP to be approximately 6.76 and 7.72,min, respectively. No interferences were shown through the retention times of morphine, buprenorphine and diazepam. With the high recovery rate of a solid-phase extraction procedure, this method was applied in analyzing plasma concentrations of seven methadone maintenance patients where R - and S -methadone and R - and S -EDDP were 233.4 ± 154.9 and 185.9 ± 136.3,ng/mL and 84.4 ± 99.4 and 37.6 ± 22.9,ng/mL, respectively. These data suggest that the present method can be applied for routine assay for plasma methadone and EDDP concentrations for patients under treatment. Copyright © 2009 John Wiley & Sons, Ltd. [source] Development and validation of a sensitive assay for the quantification of imatinib using LC/LC-MS/MS in human whole blood and cell cultureBIOMEDICAL CHROMATOGRAPHY, Issue 12 2009Jelena Klawitter Abstract We developed and validated a semi-automated LC/LC-MS/MS assay for the quantification of imatinib in human whole blood and leukemia cells. After protein precipitation, samples were injected into the HPLC system and trapped onto the enrichment column (flow 5 mL/min); extracts were back-flushed onto the analytical column. Ion transitions [M + H]+ of imatinib (m/z = 494.3 , 394.3) and its internal standard trazodone (372.5 , 176.3) were monitored. The range of reliable response was 0.03,75 ng/mL. The inter-day precisions were: 8.4% (0.03 ng/mL), 7.2% (0.1 ng/mL), 6.5% (1 ng/mL), 8.2% (10 ng/mL) and 4.3% (75 ng/mL) with no interference from ion suppression. Autosampler stability was 24 hs and samples were stable over three freeze,thaw cycles. This semi-automated method is simple with only one manual step, uses a commercially available internal standard, and has proven to be robust in larger studies. Copyright © 2009 John Wiley & Sons, Ltd. [source] A sensitive and specific liquid chromatography/tandem mass spectrometry method for determination of echinacoside and its pharmacokinetic application in ratsBIOMEDICAL CHROMATOGRAPHY, Issue 6 2009Hao Yang Abstract A rapid and sensitive method based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) for the determination of echinacoside in rat plasma was established and fully validated. A single step of liquid,liquid extraction with n -butanol was utilized. Chromatographic separation of the analyte and the internal standard (IS), chlorogenic acid, from the sample matrix was performed using a Capcell-MG C18 analytical column (100 2.0 mm × 5 µm), with a gradient of acetonitrile and water containing 0.1% acetic acid as the mobile phase. Detection was performed on a triple quadrupole tandem mass spectrometer equipped with electrospray ionization source operated in negative ion selected reaction monitoring mode. The method was linear in the concentration range 10,2500 ng/mL. The deviations of both intra- and inter-day precisions (RSD) were 7.1% and the assay accuracies were within 99.2,106.5%. Echinacoside proved to be stable during sample storage, preparation and analysis when an antioxidant solution was used. The method was successfully applied to a pharmacokinetic study in rats after an intragastric administration of echinacoside (100 mg/kg). With the lower limit of quantification at 10 ng/mL, this method proved to have sufficient selectivity, sensitivity and reproducibility for the pharmacokinetic study of echinacoside. Copyright © 2009 John Wiley & Sons, Ltd. [source] Characterization and determination of chlorophacinone in plasma by ion chromatography coupled with ion trap electrospray ionization mass spectrometryBIOMEDICAL CHROMATOGRAPHY, Issue 5 2009Xiao-kun OuYang Abstract Plasmatic chlorophacinone is commonly measured with liquid chromatographic assay, which convenient but lacks sensitivity and selectivity and usually requires ion pair reagents to reduce the chromatographic tailed peak. In this paper, a novel method using eluent generator reagent-free ion chromatography coupled with electrospray ionization ion trap mass spectrometric detection for the determination of chlorophacinone in plasma has been developed. After samples were extracted with 10% (v/v) methanol in acetonitrile and cleaned by solid-phase extraction, chromatographic separation was performed on an IonPac® AS11 analytical column (250 × 4.0 mm) using 40.0 mmol/L KOH containing 10% (v/v) methanol as organic modifier. Quantification was performed by negative electrospray ionization in multiple reaction monitoring mode. The transition m/z 373 , 201 was for the quantification ion; the transitions m/z 373 , 172 and m/z 373 , 145, as well as the isotope ions m/z 375 and m/z 203, were for the qualitative ions. All the method parameters were validated. It was confirmed that this method can be used in clinical diagnosis and forensic toxicology. Copyright © 2008 John Wiley & Sons, Ltd. [source] Comparison between micellar liquid chromatography and capillary zone electrophoresis for the determination of hydrophobic basic drugs in pharmaceutical preparationsBIOMEDICAL CHROMATOGRAPHY, Issue 1 2007S. Torres-Cartas Abstract The determination of highly hydrophobic basic compounds by means of conventional reversed-phase liquid chromatographic methods has several drawbacks. Owing to the characteristics of micellar liquid chromatography (MLC) and capillary electrophoresis (CE), these techniques could be advantageous alternatives to reversed-phase chromatographic methods for the determination of these kinds of compounds. The objective of this study was to develop and compare MLC and CE methods for the determination of antipsychotic basic drugs (amitryptiline, haloperidol, perphenazine and thioridazine) in pharmaceutical preparations. The chromatographic determination of the analytes was performed on a Kromasil C18 analytical column; the mobile phase was 0.04 m cetyltrimethylammonium bromide (CTAB), at pH 3, containing 5% 1-butanol, at a flow rate of 1 mL/min. The CE separation was performed in a fused-silica capillary with a 50 mm tris-(hydroxymethyl)-aminomethane buffer, pH 7, at an applied voltage of 20 kV, using barbital as internal stardard. The proposed methods are suitable for a reliable quantitation of these compounds in the commercial tablets and drops in terms of accuracy and precision and require a very simple pre-treatment of the samples. By comparing the performance characteristics and experimental details of the MLC and CE methods we conclude that CE seems to be slightly better than MLC in the determination of highly hydrophobic compounds in pharmaceuticals in terms of resolution and economy, taking into account that the limits of detection are not a handicap in pharmaceutical samples. Copyright © 2006 John Wiley & Sons, Ltd. [source] Liquid chromatographic/mass spectrometric assay of rabprazole in dog plasma for a pharmacokinetic studyBIOMEDICAL CHROMATOGRAPHY, Issue 11 2006Shao Feng Abstract In order to evaluate the pharmacokinetic (PK) profile of rabeprazole (RA) sterile powder for injection, a rapid, sensitive and specific assay for quantitative determination of RA in dog plasma was developed and validated. After a liquid,liquid extraction procedure, samples were analyzed by liquid chromatography,electrospray ionization mass spectrometry (LC-ESI-MS) using omepazole as the internal standard (IS). The analyte and IS was chromatographed on a ZORBAX Extend-C18 analytical column (50 × 2 mm i.d, 5 µm, Agilent Technologies, USA). The assay was linear in the range 1,2000 ng/mL. The lower limit of quantification of RA was 1 ng/mL. The recovery of RA was greater than 70%. The within- and between-batch accuracy was 102.7,107.4% and 103.5,105.7%, respectively. The plasma samples for the PK study were collected at defined time points during and after an intravenous injection (1 mg/kg) to beagle dogs and analyzed by LC-ESI-MS method. The PK parameters, such as half-life, volume of distribution, total clearance and elimination rate constant, were determined. The PK profile of RA gave insights into the application in the clinics. Copyright © 2006 John Wiley & Sons, Ltd. [source] Determination of didanosine in maternal plasma, amniotic fluid, fetal and placental tissues by high-performance liquid chromatography,tandem mass spectrometryBIOMEDICAL CHROMATOGRAPHY, Issue 6-7 2006T. Nicole Clark Abstract A rapid and efficient high-performance liquid chromatography (HPLC)-tandem mass spectrometry method for the determination of didanosine concentrations in maternal rat plasma, amniotic fluid, placental and fetal tissue samples has been developed and validated. Tissue samples were homogenized in optima water and centrifuged. The supernatant was subjected to solid-phase extraction (SPE) prior to analysis. Plasma and amniotic fluid samples were extracted without pretreatment. An Agilent 1100 Series HPLC coupled with a Micromass Quattro II triple quadrupole mass spectrometer was used for all analyses. Chromatographic resolution was achieved on a Nova-Pak phenyl analytical column (2.0 × 150 mm, 4 µm particle size) equipped with a Phenomenex Security-guard phenyl guard cartridge (2.0 × 4.0 mm) using 60% methanol in 10 mm ammonium acetate buffer mobile phase for all matrices at a flow rate of 0.15 mL/min. The method yields retention times of 2.9 min for didanosine and 3.0 min for the internal standard, stavudine. Limits of detection were 1 ng/mL for all matrices. Recoveries were 70% or greater for both compounds in the different matrices. Within- and between-run precision (%RSD) and accuracy (%error) was less than 15% for all matrices. Copyright © 2006 John Wiley & Sons, Ltd. [source] Determination of antiviral nucleoside analogues AM365 and AM188 in perfusate and bile of the isolated perfused rat liver using HPLCBIOMEDICAL CHROMATOGRAPHY, Issue 3 2006Jiping Wang Abstract Development, validation and application of an HPLC assay for new antiviral nucleoside analogues AM365 and AM188 in isolated perfused rat liver perfusate and bile were performed. An analytical column (Phenosphere-NEXT, 250 × 4.6 mm, C18, 4 µm, Phenomenex) was used in tandem with a guard column (4 × 3 mm, C18, Phenomenex) and operated at 25°C. The mobile phase [methanol:10 mmol/L sodium orthophosphate buffer (pH 7.0), 15:85, v/v] was pumped at 1 mL/min. The signal from a diode array detector was collected from 190 to 300 nm. The chromatogram was processed at 220 and 252 nm for AM365 and AM188, respectively. The HPLC method was validated by six intraday and seven interday runs. Standard curves were linear in the range 0.125,8.00 µg/mL for AM365 and AM188, and the lower limit of quantification for AM365 and AM188 was 0.125 µg/mL. Mean interday precision and accuracy of IPL perfusate quality control samples were within 8.8%, and mean intraday precision and accuracy were within 13.1%. The assay has been successfully used in the study of metabolism and disposition of AM365 in the isolated perfused rat liver. Copyright © 2005 John Wiley & Sons, Ltd. [source] High-throughput determination of atrasentan in human plasma by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometryBIOMEDICAL CHROMATOGRAPHY, Issue 9 2005Perry G. Wang Abstract Atrasentan (A-147627) is an endothelin antagonist receptor being developed at Abbott Laboratories for the treatment of prostate cancer. A quick and sensitive method for the determination of atrasentan in human plasma has been developed and validated using high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. A dual-column, single mass spectrometer system is used to provide a reliable and routine means to increase sample throughput. The analytical method involves liquid,liquid extraction and internal standard (A-166790). The plasma samples and internal standard are acidified with 0.3 m hydrochloric acid prior to being extracted into 1:1 (v[sol ]v) hexanes,methyl t -butyl ether. The organic extract was evaporated to dryness using heated nitrogen stream and reconstituted with mobile phase. Atrasentan and internal standard were separated with no interference in a Zorbax SB-C18 analytical column with 2.1 × 50 mm, 5 µm, and a Zorbax C8 guard column using a mobile phase consisting of 50:50 (v:v) acetonitrile,0.05 m ammonium acetate, pH 4.5, at a flow rate of 0.30 mL[sol ]min to provide 4 min chromatograms. For a 250 µL plasma sample volume, the limit of quantitation was approximately 0.3 ng[sol ]mL. The calibration was linear from 0.30 to 98.0 ng[sol ]mL (r2 > 0.995). A significant advantage of the method is the ability to employ parallel HPLC separations with detection by a single MS[sol ]MS system to provide sensitivity and selectivity sufficient to achieve robust analytical results with a lower limit of quantitation of 0.30 ng[sol ]mL and high throughput. Copyright © 2005 John Wiley & Sons, Ltd. [source] Rapid high-performance liquid chromatographic measurement of buspirone in human plasma after overdoseBIOMEDICAL CHROMATOGRAPHY, Issue 9 2004F. Péhourcq Abstract For toxicological purposes, a rapid reversed-phase high-performance liquid chromatographic method was developed for the determination of the anxiolytic drug, buspirone, in human plasma. A liquid,liquid procedure was used to extract this compound from plasma in the presence of an internal standard, quinupramine. The analysis was performed on a Spherisorb® S5 C8 analytical column with UV detection at 240 nm. No endogenous compounds were found to interfere. A linear response was observed over the concentration range 5,100 ng/mL. A good accuracy (bias ,7.9%) was achieved for all quality controls, with intra-day and inter-day variation coef,cients equal or less than 7.6%. The limit of quanti,cation was 5 ng/mL. Stability of buspirone in plasma stored at different temperatures was checked. This rapid method (run time <12 min) was used to manage an acute poisoning involving buspirone. Copyright © 2004 John Wiley & Sons, Ltd. [source] Application of liquid chromatography,Tandem mass spectrometry method for the analysis of new nonselective ,-adrenergic blocker 1-(1- H -indol-4-yloxy)-3-{[2-(2-methoxy phenoxy)ethylo]amino}propan-2-ol (2F109) in rat plasmaCHIRALITY, Issue 7 2007Maria Walczak Abstract A sensitive and specific liquid chromatography electrospray ionization,tandem mass spectrometry method for the enantioselective determination of the novel ,-adrenolytic compound, 1-(1- H -indol-4-yloxy)-3-{[2-(2-methoxyphenoxy)ethylo]amino} propan-2-ol, in rat plasma has been developed and validated. Chromatography was performed on a reversed-phase Chiralcel OD-RH analytical column (150 × 4.6 mm, 5 ,m, Daicel Chemical Industries, Tokyo, Japan) with isocratic elution using a mobile phase containing acetonitrile and water with 0.01% formic acid. Detection was achieved by an Applied Biosystems MDS Sciex (Concord, Ontario, Canada) API 2000 triple quadrupole mass spectrometer. Electrospray ionization (ESI) was used for ion production. The limit of detection in the MRM mode was found to be 1.25 ng/ml. The limit of quantification of both enantiomers was 2.5 ng/ml. The precision and accuracy for both intra- and inter-day determination of 2F109 enantiomers ranged from 2.6 to 12% and from 89.1 to 107.1%. This analytical method allowed us to carry out pharmacokinetic studies in rats. Our findings demonstrate that 2F109 shows stereoselective disposition in rat plasma after i.v. administration. The terminal half-lives of (+)-(R)-2F109 and (,)-(S)-2F109 were 33.5 and 42.6 min, respectively. The AUC0,inf of (+)-(R)-2F109 exceeded that of (,)-(S)-2F109. Chirality, 2007. © 2007 Wiley-Liss, Inc. [source] | ||||||||||