Development and validation of a stability-indicating method for the simultaneous estimation of sofosbuvir and ledipasvir by RP-HPLC

Abstract

A stability-indicating reversed-phase high-performance liquid chromatography method was developed and validated for simultaneous quantification of sofosbuvir and ledipasvir in tablets. The chromatographic separation was done in an isocratic mode using the Discovery C18 (250×4.6 mm, 5 µ particle size) column. The mobile phase 0.1 % ortho phosphoric acid and acetonitrile 45:55 (% v/v) at a flow rate of 1.0 ml/min and at ambient temperature was used. The wavelength used for detection was 270 nm. The retention time for sofosbuvir was found to 2.08 min and that of ledipasvir was 3.06 min. Sofosbuvir and ledipasvir were linear in the concentration ranges of 100 to 600 µg/ml and 22.5 to 135 µg/ml, respectively. The developed method was validated and found to be accurate, specific and robust. Both the drugs were subjected to the stressed conditions like acidic, basic, oxidative, photolytic and thermal conditions. The degradation results were found satisfactory. This method could be applied for the simultaneous estimation of sofosbuvir and ledipasvir in tablets.