Validation of a simple HPLC method for DRF-4848, a novel COX-2 inhibitor suitable for pharmacokinetic application in rats

✍ Scribed by Raja Reddy Kallem; Ravi Kumar Trivedi; Ramesh Mullangi; Nuggehally R. Srinivas

• Publisher: John Wiley and Sons
• Year: 2006
• Tongue: English
• Weight: 141 KB
• Volume: 20
• Category: Article
• ISSN: 0269-3879
• DOI: 10.1002/bmc.674

✦ Synopsis

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 µg/mL. The lower limit of quantification (LLOQ) of DRF-4848 was 0.1 µg/mL. The inter-and intra-day precisions in the measurement of quality control (QC) samples, 0.1, 0.3, 8.0 and 15.0 µg/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.