RP-HPLC Method for Simultaneous Determination of Sofosbuvir and Ledipasvir in Tablet Dosage Form and Its Application to In Vitro Dissolution Studies

Abstract

A reversed-phase high-performance liquid chromatographic method was developed for the simultaneous determination of sofosbuvir and ledipasvir in tablet dosage form. The analysis was performed on Luna analytical column 250 × 4.6 mm, 5 µm, octyl silica packing (Si–[CH2]7–CH3) C8, using ammonium acetate buffer solution pH 7.0 and acetonitrile 35:65 % v/v as mobile phase at flow rate of 0.7 mL min−1 for isocratic elution. Detection of sofosbuvir and ledipasvir was performed on a UV detector at 245 nm. The retention times of sofosbuvir and ledipasvir were 4.468 ± 0.013 min and 8.242 ± 0.012 min, respectively, and the total run time was 20 min. The method was validated according to the requirements of the United States Pharmacopeia (category I). The overall recovery of both analytes was 100 ± 1 %; the relative standard deviation for precision and intraday precision was less than 2.0 %. The method was linear with correlation coefficient (r) >0.9999, limits of detection 0.485 and 0.175 µg mL−1, and limits of quantification was 1.619 and 0.586 µg mL−1 for sofosbuvir and ledipasvir, respectively. The method was successfully applied to the assay and in vitro dissolution studies of sofosbuvir and ledipasvir in tablet dosage form.