The aim of this present study is to develop an accurate, precise and linear liquid chromatographytandem mass spectrometry (LC-MS/MS) method for the simultaneous estimation of sofosbuvir and velpatasvir in the pharmaceutical dosage form. The chromatographic system employs a Zorbax C18 column (50 x 4.6mmx 3µ) using the mobile phase (10mM) ammonium formate: acetonitrile in the proportion of 20:80, v/v, delivered at a flow rate of 0.8 ml/min. The developed method resulted in retention of 1.248 min and 1.530 min for sofosbuvir and Velpatasvir and Sofosbuvir produced precursor ion [M+H] at M/Z in 429.46 and corresponding product ion at M/Z in 283.29 and Velpatasvir produced a protonated precursor ion [M+H] at M/Z in 519.45 and corresponding product ion at M/Z in 216.16. Sofosbuvir and velpatasvir exhibited linear relationship (r2 > 0.99) over the analytical range 0.2-2000 ng/ml and 5-2000. The percentage recovery was found to be in the range of 96.6‐99.9% for both sofosbuvir and velpatasvir. The limit of detection (LOD) was found to be 0.1 ng/ml and 1ng/ml for sofosbuvir and velpatasvir and quantification at 0.2 ng/ml and 5 ng/ml for sofosbuvir and velpatasvir. An accurate, precise and linear method was developed and validated for the quantitative estimation of sofosbuvir(400mg) and Velpatasvir(100mg) combination tablet as per ICH guidelines and hence it can be used for the routine analysis in various pharmaceutical industries.

Sofosbuvir, Velpatasvir, LC-MS/MS, ICH.