Phytochemical Profiling of Withania somnifera for Qualitative and Quantitative Study of Bioactive Component Withaferin a by High-Performance Liquid Chromatography

Over the past few years, the attention of people has changed from man-made (synthetic) to natural medicines. Withania somnifera’s phytochemical analysis has identified the presence of withanolides, which are pharmacologically active steroidal lactones. The two main withanolides that were isolated from Withania somnifera in India were withanolide D and withaferin A, both of which demonstrated cytotoxic and anticancer effects. The present investigation aimed to evolve a standard phytochemical profiling of Withania somnifera for the presence of withaferin A. The dried root sample was minced and ground with a mechanical grinder. The bioactive reference standard withaferin A was used. Methanol was used to extract the active fraction. By comparing the spectra of standard withaferin A with the root powder of Withania somnifera extracted using methanol, the peak purity of withaferin A was determined. The calibration curve was established for peak area vs concentration of Withaferin A. From the standard stock solution, six different concentrations (0.0 ppm, 1.25 ppm, 250 ppm, 5.0 ppm, 10.0 ppm, and 20.0 ppm) were used for preparing six six-point calibration curves. The comparative analysis of standard withaferin A and the sample has been determined. The sample (Withania somnifera root powder) and standard solutions were run in methanol mobile phase systems. It was found that after running the sample and standard withaferin A, it had a good resolution, a sharp and symmetrical peak at retention times 13.685 min. and 13.600 min. respectively. Estimation of Withaferin A can be used by the pharmaceutical industry as an appropriate bioactive marker compound for phytochemical profiling and quality control of Withania somnifera. Phytochemical profiling of Withania somnifera is essential to assess the quality, purity, efficacy, and safety of the bioactive component (withaferin A). The HPLC chromatogram of Withania somnifera root powder extract corresponding to standard Withaferin A was shown at a retention time of 13.685 min, at 227 nm wavelength. The quantitative evaluation of Withaferin A present in the sample was 0.10%.