Development And Validation of RP-HPLC Method for The Estimation of Ponesimod in Bulk and Tablet Dosage Form

The pharmaceutical industry demands precise and robust analytical methods to ensure drug quality, safety, and efficacy. This thesis presents the development and validation of a reliable Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method for estimating Ponesimod, a selective sphingosine-1-phosphate receptor modulator, in bulk and tablet dosage forms. Method development focused on optimizing chromatographic parameters such as mobile phase composition and flow rate to achieve high resolution and minimal tailing. The final method demonstrated excellent linearity across the 10–30 µg/mL concentration range, with a correlation coefficient (r²) > 0.999. System suitability tests confirmed acceptable retention time, theoretical plates, and tailing factor. Accuracy was validated through recovery studies, with results ranging from 98% to 102%, while precision studies showed %RSD values below 2%, affirming method reproducibility. The method's sensitivity was established with LOD and LOQ values of 1.09 µg/mL and 3.29 µg/mL, respectively. Robustness was proven by varying analytical conditions, and forced degradation studies confirmed its stability-indicating capability under acid, base, oxidative, thermal, and photolytic stress. Overall, the validated method complies with ICH guidelines and is suitable for routine analysis of Ponesimod in pharmaceutical formulations..