DEVELOPMENT AND VALIDATION STUDY OF RP-HPLC METHOD FOR QUANTITATIVE ANALYSIS OF SOFOSBUVIR

Abstract

A precise, accurate, time saving and cost effective reverse phase high performance liquid chromatographic method was developed for the quantitative determination of sofosbuvir in bulk and in pharmaceutical tablet dosage form. This method was validated with respect to the specificity, accuracy, precision, limits of detection, linearity etc.  The Separation was carried on ZORBAX Eclipse Plus C18 column (Length: 150 mm, diameter: 4.6 mm, and particle size of 5 micron) by using 0.05 M phosphoric acid and acetonitrile (66:34% v/v) as mobile phase @ flow rate of 2 mL/min. Total run time of the analysis was kept 6 min and the detection was carried out at the wavelength of 265 nm by using photodiode array detector. Symmetrical peak for Sofosbuvir was obtained with tailing factor of 1.40 at retention time 2.74min. This method gave linear response in the concentration range of 0.003125 mg/mL to 1 mg/mL and value of correlation coefficient (R²) is 0.9999. Precision of method was calculated in terms of repeatability and intermediate precision with relative standard deviation < 2. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.002754 mg/mL and 0.009181 mg/mL respectively. The accuracy of method was calculated in term recovery percentage by spiking 50%, 100% and 150% amount of sofosbuvir in test concentration and result were observed accurate. A verification study for the determination of sofosbuvir in pharmaceutical tablet dosage form was also performed and results were observed very precise and accurate. Hence, the developed method can be safely used for the quantitative determination of sofosbuvir.