生物分析与生物医学杂志

UPLC Method for Simultaneous Estimation of Ledipasvir and Sofosbuvir in Bulk and Dosage Forms and Their Stress Degradation Studies

Abstract

K Pushpa Kumari and D Gowri Sankar

Ultra-performance liquid chromatographic (UPLC) method, for ledipasvir and sofosbuvir determination in bulk mixtures, and in tablets was developed and validated. The determination of the drugs, ledipasvir and sofosbuvir was carried out applying BEH C18 column (50 mm × 2.1 mm i.d, 1.6 μm particle size), with UV detector at λ max of 220 nm. The mobile phase applied for the current study, composed of two solvents, A (0.01 N w/v potassium di-hydrogen orthophosphate buffer of pH 3.5 modified with dilute orthophosphoric acid) and B (acetonitrile) in the ratio of 50:50 v/v. The isocratic mobile phase was moved at a flow rate of 0.3 ml/min. The validation study with proportionate to selectivity, linearity, robustness, precision, accuracy, stability, (LOD) limit of detection and (LOQ) limit of quantification, was performed utilizing the ICH Guidelines. Ledipasvir and Sofosbuvir displaying a linear response between 22.5-135 μg/ml for Ledipasvir 100-600 μg/ml for Sofosbuvir, with correlation coefficient (R2) 0.9997 and 0.998 for Ledipasvir and Sofosbuvir respectively. The % recovery for Ledipasvir and Sofosbuvir was 99.72 ± 0.61% and 100.21 ± 0.68 respectively. The LOD and LOQ values for Ledipasvir and Sofosbuvir were obtained to be 0.31 and 0.95 μg/ml and 1.29 and 3.91 μg/ml, respectively. The method also exhibits good robustness for different chromatographic conditions like mobile phase, temperature and flow rate. So, the approach can be successfully implemented for the quantitative analysis of ledipasvir and sofosbuvir in the (Q.C) Quality Control of in-house developed tablets, and the same can be exercised for the industrial use.