Estimation of Withaferin-A by HPLC and standardization of the Ashwagandhadi lehyam formulation

Standardization is an important measurement for ensuring the quality control of herbal drugs. It has become essential to develop reliable, specific and sensitive quality control methods. Ashwagandhadi lehyam is an important Ayurvedic formulation containing Withania somnifera L., as one of the prime ingredient of formulation. The present study was undertaken to develop standardization parameters for Ashwagandhadi lehyam. Evaluation of various standardization parameters like organoleptic characters, Physico-chemical evaluation, HPTLC finger print profiling along with dominant bioactive markers and estimation of bioactive markers Withaferin-A by HPLC. The R_f value of Withaferin-A 0.35 and Withanolide-A 0.45 is in formulation and reference standards were found comparable under UV light at 254 nm and 540 nm. The HPLC chromatogram of Ashwagandhadi lehyam and standard Withaferin-A showed at Rt of 5.015 and 5.050 min. The percentage of Withaferin-A was 0.092% present in Ashwagandhadi lehyam formulation. Bioactive markers are characteristic to the ingredients or botanicals to identify the presence of ingredients in formulation easily. The presence of bioactive markers is possible and its verification through the HPTLC fingerprint profile and quantification of biomarker by HPLC are the best ways to identify evaluate the quality of the finished formulation in the course of development of a standardization protocol for quality control of Ayurvedic formulation.