Objective: A strategic quantitative analysis of Ledipasvir (LEDI) and Sofosbuvir (SOF) has been developed in Bulk and pharmaceutical (Tablet) dosage form with a reliable, Precise and sensitive First-Order Derivative spectrophotometric and RPHPLC methods. Methods: Derivative method was developed firstly using methanol as a solvent, obeying the Beer’s Law in a concentration range from 2-12 μg/ml and 8-48 μg/ml for LEDI and SOF showing the ZCP at 284 nm and 257 nm respectively. Secondly, better chromatographic separation and estimation was attained with a sample Pre-treated automated and analysed using a BDS-C18 (250 mm × 4.6 mm, 5µm) column and with mobile phase composition of Methanol: Acetonitrile: (1%) Ammonium acetate (50:20:30 % v/v/v) at 1 ml/min flow rate with an UV detection at 281 nm. Developed methods were validated rendering to ICH Q2 (R1) guidelines.
Results: Linear Calibration curve was obtained covering the whole of the concentration range with regression coefficient value 0.998 and 0.999 for LEDI and SOF in the Spectrophotometric method. Rectilinear relationship with acceptable regression coefficient 0.9987 and 0.997 was found over the concentration ranging from 1-11 μg/ml for LEDI and 4-44 μg/ml for SOF respectively in the RP-HPLC method. SOF and LEDI were detected at retention time of 5.388 and 8.085 min respectively. The mean percentage recoveries were in the range of 98.51 – 101.6 % and 99.11 – 101.22 % for LEDI and SOF. Conclusion: The results indicate that the developed RP-HPLC and UV spectrophotometric methods are suitable for the routine quality control testing of marketed tablet formulations.
