Analytical Method Development and Validation of Withaferin A and Piperine in Capsule Dosage Form

Abstract

HPLC method has been developed for simultaneous estimation of Withaferin A and Piperine in capsule dosage form. The method is uncomplicated, precise, accurate and robust. The separation was executed by Phenomenex C-18 (250 × 4.6 mm, 5µm particle size) column and Acetonitrile: water (70: 30 %v/v) as a mobile phase with the flow rate 1ml/min. Detection was carried out at wavelength of 230 nm. The retention time for Withaferin A and Piperine were found to be 3.77 and 4.55 respectively. Linearity complied for range of 5-15 μg/ml for Withaferin A and 25-75 μg/ml for Piperine. The accuracy of the method was judged by recovery studies. The developed method was validated as per the ICH guidelines parameters like linearity, precision, accuracy, robustness, LOD and LOQ. The results were validated as per ICH Q2 R1 guideline and were adequate. This method can be successfully employed for simultaneous estimation of Withaferin A and Piperine in bulk drugs and formulation