Stability-Indicating HPLC Method for Simultaneous Estimation of Sofosbuvir and Ledipasvir

The optimization of chromatographic conditions for the analysis of Ledipasvir and Sofosbuvir was successfully achieved using a Hibar C18(250×4.6mm)5µm column using 1% O-Phosphoric acid (pH 6.4): Acetonitrile (30:70 v/v) at a flow rate of 0.8 ml/min, ensuring excellent peak shape and resolution. System suitability tests confirmed compliance with essential parameters, including resolution, tailing factor, and theoretical plate count, ensuring reliability. The retention time of ledipasvir and sofosbuvir were found to be 7.1min and 3.7 min, respectively. During force degradation, drug product was exposed to hydrolysis (acid and base hydrolysis), oxidation, thermal degradation and photo degradation. Both the drugs were not degraded under thermal, oxidative, photolytic, acid and neutral hydrolytic conditions, but Sofosbuvir showed degradation under alkaine hydrolytic condition with a retention time 2.8 min and 4.1 min, respectively. The degraded products of Ledipasvir and Sofosbuvir were well resolved from the individual bulk drug response. The specificity of the method was confirmed by peak purity profile of the resolved peaks. In conclusion, the developed chromatographic method for Ledipasvir and Sofosbuvir is precise, accurate, linear, sensitive, and robust, making it suitable for routine pharmaceutical analysis and quality control. The validation results confirm compliance with regulatory standards, ensuring reliable identification and quantification of these antiviral drugs. Given its high stability and reproducibility, this method provides an efficient and dependable approach for analyzing Ledipasvir and Sofosbuvir in pharmaceutical formulations.