Abstract

ANALYTICAL METHOD VALIDATION FOR THE DETERMINATION OF RELATED SUBSTANCE IN SOFOSBUVIR 400MG TABLET BY USING REVERSE PHASE HPLC TECHNIQUE

Irshad Alam*, Junaid Ali and Kashif Ahmed

ABSTRACT

The present work is a development and validation of a method forquantitative measurement of degradation of active pharmaceuticalingredient (API) Sofosbuvir i.e. SF10, SF11, SF12, SF13 in the drugproduct by using the Reverse Phase High Performance LiquidChromatography (HPLC) analytical method with UV detection systemfor the qualitative and quantitative determination. Chromatographicseparation was achieved by using gradient method on L1 column as aStationary Phase with the specification of 4.6mm × 10-cm, 3-μm, andPhosphate buffer + Acetonitrile as a Mobile Phase, with differentcomposition at flow rate 1.0mL/minute, at 260nm UV detection, and at20μL sample volume. The method found linear with an adequate level of precision andaccuracy over the concentration range of 0.5μg/mL to 995.98μg/mL. The percentage RSD forprecision and accuracy of the method was found to be less than 1%. The method wasvalidated according to the International Conference on Harmonization (ICH) guidelines withrespect to specificity, linearity, accuracy, precision, limit of detection (LOD), and limit ofquantitation (LOQ). The developed method successfully applied for analysis of relatedsubstances of drug product “Sofosbuvir”.

Keywords: HPCL, gradient elution, simultaneous, Sofosbuvir