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Absolute Recovery (absolute + recovery)
Selected AbstractsValidation of a simple HPLC method for DRF-4848, a novel COX-2 inhibitor suitable for pharmacokinetic application in rats,BIOMEDICAL CHROMATOGRAPHY, Issue 12 2006Raja Reddy Kallem Abstract For pharmacokinetic and toxicokinetic purpose a simple HPLC-UV method has been developed and validated for the estimation of DRF-4848, a novel COX-2 inhibitor in rat plasma. A liquid,liquid extraction was used to extract DRF-4848 and internal standard (IS, DRF-4367) from rat plasma. The analysis was performed on a C18 column with UV detection at 285 nm. The isocratic mobile phase, 0.01 m potassium dihydrogen ortho phosphate (pH 3.2) and acetonitrile (50:50, v/v) was run at a flow rate of 1 mL/min. The retention times of DRF-4848 and IS were 6.8 and 11.2 min, respectively. Absolute recovery for analyte and IS was >80% from rat plasma. A linear response was observed over a concentration range 0.1,20 mg/mL. The lower limit of quantification (LLOQ) of DRF-4848 was 0.1 mg/mL. The inter- and intra-day precisions in the measurement of quality control (QC) samples, 0.1, 0.3, 8.0 and 15.0 mg/mL, were in the range 1.74,8.70% relative standard deviation (RSD) and 0.75,8.43% RSD, respectively. Accuracy in the measurement of QC samples was in the range 93.29,116.51% of the nominal values. Analyte and IS were stable in the battery of stability studies viz., benchtop, autosampler, long-term and freeze/thaw cycles. Copyright © 2006 John Wiley & Sons, Ltd. [source] Development and validation of a sensitive ELISA for quantitation of Grastim® (rhG-CSF) in rat plasma: application to a pharmacokinetic study,BIOMEDICAL CHROMATOGRAPHY, Issue 9 2006Chandrasekar Nirmala Abstract Grastim® is bacterially produced recombinant counterpart of human granulocyte colony stimulating factor (G-CSF). It has biological activity similar to that of endogenous G-CSF. In the present work a sensitive, accurate, precise and enzyme-linked immunosorbent assay (ELISA) for the quantitation of G-CSF in rat plasma was developed and validated. The ELISA method employed a technique in which anti-human-G-CSF was adsorbed onto 96-well maxisorp plates and used to capture the G-CSF in rat plasma samples. The captured G-CSF was then detected using streptavidin-HRP amplification system. Absolute recovery was >90% from rat plasma. The validation includes assessments of method accuracy and precision, range of reliable response, lower limit of quantitation (LLOQ), storage stability (30 days) in rat plasma and assay specificity. The standard curve for G-CSF was linear (R2 > 0.996) in the concentration range 4.88,625 pg/mL. The LLOQ was established at 4.88 pg/mL. The inter- and intra-day precisions in the measurement of quality control (QC) samples, 15, 250 and 500 pg/mL, were in the range 3.00,8.66% relative standard deviation (RSD) and 1.03,4.69% RSD, respectively. Accuracy in the measurement of QC samples was in the range 87.28,110.79% of the nominal values. The assay shows dilutional linearity and specificity. Stability of G-CSF was established for 30 days at ,80°C and through three freeze,thaw cycles. The validated assay was successfully employed for the assessment of pharmacokinetic disposition of G-CSF in rats. Copyright © 2006 John Wiley & Sons, Ltd. [source] HPLC method for determination of DRF-4367 in rat plasma: validation and its application to pharmacokinetics in Wistar rats,BIOMEDICAL CHROMATOGRAPHY, Issue 8 2004Ramesh Mullangi Abstract A speci,c, accurate, precise and reproducible high-performance liquid chromatography (HPLC) method was developed for the estimation of DRF-4367, a novel cyclooxygenase-2 inhibitor in rat plasma. The assay procedure involved simple liquid/liquid extraction of DRF-4367 and internal standard (IS, celecoxib) from plasma into dichloromethane. The organic layer was separated and evaporated under a gentle stream of nitrogen at 40°C. The residue was reconstituted in the mobile phase and injected onto a Kromasil KR 100-5C18 column (4.6 × 250 mm, 5 µm). The mobile phase consisting of 0.01 m potassium dihydrogen ortho -phosphate (pH 3.2) and acetonitrile (40:60, v/v) was used at a ,ow rate of 1.0 mL/min. The eluate was monitored using an UV detector set at 247 nm. The ratio of peak area of analyte to IS was used for quanti,cation of plasma samples. Nominal retention times of DRF-4367 and IS were 6.6 and 11.2 min, respectively. The standard curve for DRF-4367 was linear (r2 > 0.999) in the concentration range 0.1,20 µg/mL. Absolute recovery was >86% from rat plasma for both analyte and IS. The lower limit of quanti,cation of DRF-4367 was 0.1 µg/mL. The inter- and intra-day precisions in the measurement of quality control samples, 0.1, 0.3, 8.0 and 15.0 µg/mL, were in the range 6.93,9.34% relative standard deviation (RSD) and 0.48,6.59% RSD, respectively. Accuracy in the measurement of QC samples was in the range 91.24,109.36% of the nominal values. Analyte and IS were stable in the battery of stability studies, viz. benchtop, autosampler and freeze,thaw cycles. Stability of DRF-4367 was established for 1 month at ,80°C. The application of the assay to a pharmacokinetic study in rats is described. Copyright © 2004 John Wiley & Sons, Ltd. [source] Simple, sensitive and rapid liquid chromatographic/electrospray ionization tandem mass spectrometric method for the quantification of lacidipine in human plasmaJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 7 2004N. V. S. Ramakrishna Abstract A simple, sensitive and rapid liquid chromatographic/electrospray ionization tandem mass spectrometric method was developed and validated for the quantification of lacidipine in human plasma using its structural analogue, amlodipine, as internal standard (IS). The method involves a simple single-step liquid,liquid extraction with tert -butyl methyl ether. The analyte was chromatographed on an Xterra MS C18 reversed-phase chromatographic column by isocratic elution with 20 mM ammonium acetate buffer,acetonitrile (10 : 90, v/v; pH 6) and analyzed by mass spectrometry in the multiple reaction monitoring mode. The precursor to product ion transitions of m/z 456.4 , 354.4 and m/z 409.3 , 238.3 were used to measure the analyte and the I.S., respectively. The chromatographic run time was 1.5 min and the weighted (1/x2) calibration curves were linear over the range 0.1,25 ng ml,1. Lacidipine was sensitive to temperature in addition to light. The method was validated in terms of accuracy, precision, absolute recovery, freeze,thaw stability, bench-top stability and re-injection reproducibility. The limit of detection and lower limit of quantification in human plasma were 50 and 100 pg ml,1, respectively. The within- and between-batch accuracy and precision were found to be well within acceptable limits (<15%). The analyte was stable after three freeze,thaw cycles (deviation <15%). The average absolute recoveries of lacidipine and amlodipine (IS) from spiked plasma samples were 51.1 ± 1.3 and 50.3 ± 4.9%, respectively. The assay method described here could be applied to study the pharmacokinetics of lacidipine. Copyright © 2004 John Wiley & Sons, Ltd. [source] Optimization of a novel headspace,solid-phase microextraction,gas chromatographic method by means of a Doehlert uniform shell design for the analysis of trace level ethylene oxide residuals in sterilized medical devicesBIOMEDICAL CHROMATOGRAPHY, Issue 6 2009Michael P. DiCicco Abstract Medical devices sterilized by ethylene oxide (EtO) retain trace quantities of EtO residuals, which may irritate patients' tissue. Reliably quantifying trace level EtO residuals in small medical devices requires an extremely sensitive analytical method. In this research, a Doehlert uniform shell design was utilized in obtaining a response surface to optimize a novel headspace,solid-phase microextraction,gas chromatographic (HS-SPME-GC) method developed for analyzing trace levels of EtO residuals in sterilized medical devices, by evaluating sterilized, polymer-coated, drug-eluting cardiovascular stents. The effects of four independent experimental variables (HS-SPME desorption time, extraction temperature, GC inlet temperature and extraction time) on GC peak area response of EtO were investigated simultaneously and the most influential experimental variables determined were extraction temperature and GC inlet temperature, with the fitted model showing no evidence of lack-of-fit. The optimized HS-SPME-GC method demonstrated overall good linearity/linear range, accuracy, repeatability, reproducibility, absolute recovery and high sensitivity. This novel method was successfully applied to analysis of trace levels of EtO residuals in sterilized/aerated cardiovascular stents of various lengths and internal diameter, where, upon heating, trace EtO residuals fully volatilized into HS for extraction, thereby nullifying matrix effects. As an alternative, this novel HS-SPME-GC method can offer higher sensitivity compared with conventional headspace analyzer-based sampling. 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