Hplc Method Development and Validation for Determination of Raltegravir in Blood Plasma

A simple, precise and specific reverse phase high performance liquid chromatographic method has been developed and validated for the determination of raltegravir in blood. The HPLC separation was carried out by reverse phase chromatography on Shimadzu HPLC system consisted of Welchrom 5μ C18 column (250 X 4.6 mm), SPD 10A UV detector and LC 10AD Pumps. Rheodyne injector with 20 μl loop with a mobile phase composed of Acetonitrile : Water pH 2.5 (70:30) at a flow rate 1.0 mL/min. Naproxen was used as internal standard in the determination. The retention time of raltegravir and internal standard was found to be 5.7 and 10.2 respectively. The detection was monitored at 251 nm. The calibration curve for raltegravir was linear from 0.1-10μg/mL with correlation coefficient of 0.999. The interday and intraday precision was found to be within limits. The method was validated as per the guidelines.