Comparative Study of Withaferin A and Withanolide A in Different Cultivators of Withania Somnifera by RP-HPLC Method

Abstract

Introduction: Ashwagandha (Withaniasomnifera) is a significant and highly valued medicinal plant in various indigenous system of medicine (Ayurveda, siddha and unani).In the present study an attempt is made to develop RP-HPLC method for the estimation of important chemical constituents like Withaferin-A and Withanolide-A. Also, qualitative and quantitative evaluation of Withaferin A and Withanolide collected from different cultivars. This method was reproducible, accurate and buffer free. Several samples collected from different parts of India and analyzed by HPLC using PDA detector. MATERIAL AND METHOD: Withaferin A and Withanolide A, India used as a reference standards for method development and analysis for W. Somnifera samples. Methanol, Acetonitrile, Water used was HPLC grade. One grams of each W. Somnifera powdered root plant material was weighed in to a conical flask. Methanol was added and this was extracted by sonication. Extraction was filtered through filter paper. The samples are analyzed by HPLC above developed HPLC method. Results: The analysis was performed on PhenomenexSphereclone ODS (2)column (4.6 X 250 mm, 5 μm particle size) through gradient mode using Acetonitrile and Water as mobile phase for 50 min and spectral acquisition was performed at λmax 227 nm. Conclusion: The comparative studies done for variations of Withaferin A and Withanolide A from W. Somniferacollected from different places of India i.e., GadwalAnanthapur District of Andhra Pradesh, Kamareddy Districts of Telangana, Uttar Pradesh and CSIR-CIMAP, Research Centre, Hyderabad. Kamareddy district area sample contains more percentage of Withaferin A (0.319%) and Withanolide A (0.359%) compare to other regions.