Method Development and Validation to Estimate Sofosbuvir in Marketed preparation by UV-Spectroscopy and HPLC along with force Degradation Study

Abstract

Simple, swift, selective and accurate UV and HPLC methods were developed and validated for estimation of sofosbuvir in bulk and marketed preparation. In the UV spectroscopy method mobile phase used was methanol in 70:30 ratio with a detection wavelength of 260nm and the assay value obtained was 99.36%. The method was validated as stated by ICH in Q2 R1 guidelines in which linearity was detected from 06-30µg/ml range with regression value of 0.999. In the accuracy, precision and robustness studies RSD were below 2%. In HPLC method, Cosmosil C18 (250mm×4.6ID, Particle size: 5µ) column was utilized with methanol: water (70:30) as mobile phase, 0.9ml/min of flow rate, 260nm detection wavelength for estimation of sofosbuvir. Assay value obtained using this optimized parameters was 99.77% with the time of retention of around 4.3 minutes. HPLC method was also validated as stated by ICH in Q2 R1 guidelines in which linearity was noticed to be in the span of 10-50µg/ml with 0.999 of regression coefficient. LOD and LOQ values of the optimized method were 0.5764 and 1.7468µg/ml. In the accuracy, precision, robustness studies the value of RSD was under 2%. The optimized HPLC method was also utilized for the force degradation study, in which it was found that sofosbuvir is susceptible to oxidative, acid, alkaline, photolytic and thermal degradation. From this study it can be concluded that the developed methods can be employed in the routine analysis for sofosbuvir estimation in bulk and marketed preparation and also to determine degradation of drug.