title: Validation of a simple HPLC method for DRF-4848, a novel COX-2 inhibitor suitable for pharmacokinetic application in rats.
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 C(18) 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 microg/mL. The lower limit of quantification (LLOQ) of DRF-4848 was 0.1 microg/mL. The inter- and intra-day precisions in the measurement of quality control (QC) samples, 0.1, 0.3, 8.0 and 15.0 microg/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.
authors.affiliation: Department of Drug Metabolism and Pharmacokinetics, Discovery Research, Dr Reddy's Laboratories Ltd, Miyapur, Hyderabad-500 049, India.
authors.forename: Raja Reddy
authors.initials: RR
authors.lastname: Kallem
authors.forename: Ravi Kumar
authors.initials: RK
authors.lastname: Trivedi
authors.forename: Ramesh
authors.initials: R
authors.lastname: Mullangi
authors.forename: Nuggehally R
authors.initials: NR
authors.lastname: Srinivas
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D052246:Cyclooxygenase 2 Inhibitors / Q000097:blood / Q000493:pharmacokinetics*; D004355:Drug Stability; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013450:Sulfones / Q000097:blood / Q000493:pharmacokinetics*
chemical_list: C517299:4-(2-fluoro-4-methanesulfonylphenyl)-3-(4-fluorophenyl)-5H-furan-2-one; D052246:Cyclooxygenase 2 Inhibitors; D013450:Sulfones; D015107:4-Butyrolactone