Bioanalytical RP-HPLC method development and validation for estimation of azelnidipine and Olmesartan medoxomil in human plasma

Abstract

An accurate, rapid and simple reversed-phase high performance liquid chromatography (RP-HPLC) bioanalytical method was developed for estimation of Azelnidipine (AZP) and Olmesartan medoxomil (OLM)in human plasma. The AZP, OLM and internal standard (IS) were extracted by liquid-liquid extraction technique. Chromatographic separation was accomplished using BDS Hypersil C18, 250 mm X 4.6 mm, 5μ analytical column. The mobile phase consisted of Acetonitrile: Water, pH adjusted with ortho-phosphoric acid in the ratio 60:40 with flow rate of 1ml/min. Detection and quantification were performed by UV detector at 256 nm. The retention times were found to be 10.05±0.02, 4.3±0.02 and 6.9±0.02 for AZP, OLM and IS respectively. The linearity range was found to be 0.5 to 12 μg /ml and 1 to 15 μg /ml for AZP and OLM respectively.The method was validated as per CDER guideline, was found to be suitable for analysis in biological fluid.