Abstract

METHOD DEVELOPMENT AND VALIDATION FOR THE DETERMINATION OF RELATED SUBSTANCES IN TERIFLUNOMIDE BY RP-HPLC METHOD

Sushma G., Sridevi Pingali*, Bhagavan Raju M. and Vinutha K. Shaik Ashfaque

Abstract

A Reverse Phase High Performance Liquid Chromatographic Method (RP-HPLC) was developed and validated for the determination of related substances in Teriflunomide. The method was carried out using Zorbax SB Phenyl, 150 x 4.6 mm, 3.5μmwith mobile phase-A and mobile phase-B, where mobile phase-A consisted of potassium dihydrogen phosphate buffer and mobile phase-B consisted of Acetonitrile in the ratio 60:40 (v/v) under gradient mode with an injection volume of 10μL at flow rate of 1mL/min and the detection was carried out at 209 nm. The retention time of Teriflunomide and its related substance (RS) method were found to be 11.797min and 23.588min. This method produced linear responses for Teriflunomide and its related substances in the concentration range of 0.012-0.304% with a correlation coefficient of 0.999 and 0.999. The proposed method was validated as per ICH guidelines. The method was found to be linear, specific, precise and accurate. Therefore, the proposed method can be used for the routine analysis of drug in the process control of bulk and pharmaceutical formulation.

Keywords: RP-HPLC, Teriflunomide, Validation, related substances, pharmaceutical formulation.