Determination of a Novel COX-2 Inhibitor in Mouse Plasma: Application to Preclinical Pharmacokinetic Studies

Abstract

For preclinical study an HPLC method was developed and validated for the determination of a novel and selective cyclooxygenase-2 (COX-2) inhibitor in mouse plasma. The sample preparation procedure involved simple protein precipitation following extraction of analyte and internal standard (IS) from plasma. The separation was achieved on a Lichrospher C8 column using an isocratic mobile phase consisting of water, acetonitrile and triethylamine (48:52:0.1, v/v) as eluent with UV detection at 330 nm. A linear response was observed over a concentration range of 30-2000 ng/mL. The limits of detection and quantification were 10 and 30 ng/mL, respectively. The coefficients of variation for inter-day and intra-day assay were found to be less than 5.0 %. The assay was successfully applied to determine the pharmacokinetic parameters in mice after IV administration of the analyte. The method showed good characteristics of linearity, sensitivity, accuracy, selectivity and precision that made it suitable for preclinical pharmacokinetic studies.