RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF WITHAFERIN-A IN RANGER CAPSULE

A simple, accurate, precise and robust reverse phase High Performance Liquid Chromatographic method has been developed and validated for the estimation of Withaferin-A in polyherbal formulation namely Ranger Capsule. The chromatographic method was carried out isocratically with Phenomenex Luna C18 (2) (4.6 x 250 mm, 5 μ) and good resolution was achieved with acetonitrile: water (60: 40v/v) mobile phase. Withaferin- A was detected at retention time 4.04 min using flow rate 1.0 ml/min with UV detection at 230 nm. The calibration curve was achieved to be linear over a range of 2-20 μg/ml with regression coefficient of 0.9993. The proposed method was found accurate with 99.43 % – 100.64 % recovery. Relative standard deviation of repeatability, intra-day and inter-day variations were found to be 0.79 %, 0.12-1.55 %, 0.04-1.73 %, respectively. The Limit of Detection (LOD) and Limit of Quantification (LOQ) were observed to be 0.05 μg/ml and 0.16 μg/ml, respectively. This results shows that method was well validated. In the present study, Withaferin- A was found 80 µg per capsule. This study reveals that proposed method is quicker and cost effective for routine analysis of Withaferin- A in polyherbal dosage form