Determination of Valdecoxib in Human Plasma Using Reverse Phase HPLC

Abstract

An RP‐HPLC analytical method for estimation of valdecoxib in human plasma was developed and validated. Protein precipitation and valdecoxib extraction from plasma (200 μL) was carried out by adding 800 μL perchloric acid (5%, v/v in water) containing nimesulide as the internal standard followed by vortex mixing and centrifugation. The supernatant (20 μL) was then injected onto an ODS C18 (25 cm×4.6 mm) column from Shimadzu. The mobile phase comprised of acetonitrile and water (35: 65) with a total run time of 12 min and the wavelength of the detector was set at 244 nm. The extraction recovery of valdecoxib from plasma was >95% and the calibration curve was linear (r2 = 0.999) over valdecoxib concentrations ranging from 20 to 1400 μg/mL (n = 10). The method had an accuracy of >92% and LOD and LLOQ of 3.58 μg/mL and 13.45 μg/mL respectively. The method reported is simple, reliable, precise, accurate and has the capability of being used for determination of Valdecoxib in clinical settings.