RP-HPLC Method Development and Validation for Simultaneous Estimation of Ledipasvir and Sofosbuvir in Bulk and Tablets

P. Vamsi Reddy*1, V. Asha Ranjani2, R. Chandra Sekhar3, M. Shyam Sundar3
1Centre for Pharmaceutical Sciences, IST, JNTU, Hyderabad -500085
2MLR Institute of Pharmacy, Dundigal (v), Hyderabad-500043
3Department of Pharmaceutical analysis& Quality assurance, OU, Hyderabad-500007

A B S T R A C T
The present paper describes a new, simple, precise, and accurate RP-HPLC method for simultaneous estimation of ledipasvir and sofosbuvir in bulk and formulation. Separation was performed on Kromacil-100 HD,C18 column (15cm x 4.6mm i.e., 5 μm) at ambient temperature. The mobile phase consisted of ammonium acetate buffer (pH 6.0), acetonitrile and methanol in the ratio of 30:40:30 (v/v) at a flow rate of 1.0 ml/min using isocratic pump system. Detection was carried out at wavelength 285 nm. The retention times of ledipasvir and sofosbuvir were 3.06 min and 7.45 min, respectively. The linearity was established over the concentration ranges of 50-175µg/ml with correlation coefficients of 0.998 and 0.999 for ledipasvir and sofosbuvir, respectively. The mean recoveries were found to be in the ranges of ledipasvir was 98.7-100.1% and sofosbuvir was 98.8-100.1% respectively. The proposed method has been validated as per ICH guidelines and successfully applied to the estimation of ledipasvir and sofosbuvir in their combined tablet dosage form.
Keywords: RP-HPLC, Method development, Validation, Simultaneous estimation, Robustness.