Development and Validation of Bioanalytical Method for Determination of Flurbiprofen from Human Plasma by Liquid Chromatography

A sensitive high performance liquid chromatographic method with UV detection HPLC-UV for the determination of Flurbiprofen from plasma was developed and validated. Liquid-liquid extraction was used for extracting Flurbiprofen from plasma. Diethylether:dichloromethan:isop ropanol, 3:1.5:0.5, v/v/v mixture was used as extraction solvent. The chromatographic separation of Flurbiprofen and losartan IS was carried out using reverse phase Nucleosil C18 column 150x4.6 mm, 5mm with mobile phase of 0.1 M sodium acetate:acetonitrile 65:35, v/v , pH:6.30 and UV detection at a wavelength 248 nm. The flow rate of mobile phase was 1 mL/min, injection volume was 25 ml. The calibration curve was linear within the concentration range 100- 40 000 ng/mL. The limit of quantification was 100 ng/mL with good accuracy and precision. Intraday and interday precision were less than 7.3 and 12.0 respectively and the range of accuracy was 2.5-7.3 % . Recoveries of Flurbiprofen and IS from plasma ranged from 68.1 to 72.0% for 200, 4000 and 80000 ng/ mL concentrations. The stability was assessed under a variety of conditions FDA, Bioanalytical Method Validation Guidline and found that appropriate for the quantification. The method developed has been applied successfully to pharmacokinetic study.