Development of hptlc fingerprint by simultaneous estimation of ashwagandha (withaferin-A) and glycyrrhiza(18-β-glycyrrhetinic acid) in some polyherbal over the counter marketed formulations

Trivedi Manisha N. and Santani

Abstract

A new simple, precise, rapid and selective high‐performance thin‐layer chromatographic (HPTLC) method has been developed and validated for the simultaneous determination of withaferin A and 18-β-Glycyrrhetinic acid in Ayurvedic formulations containing Ashwagandha and Glycyrrhiza. The retention factors of withaferin A and 18-β- Glycyrrhetinic acid were 0.27 ± 0.02 and 0.72 ± 0.01 respectively. Linearity was obtained in the range of 1‐5 μg mL‐1 for withaferin A and 18-β-Glycyrrhetinic acid. The developed and validated HPTLC method was employed for standardization of three Ayurvedic formulations for their content of the two markers. Satisfactory recoveries of 98.39 to 99.77% 98.46 to 99.60% were obtained for withaferin A and 18-β-Glycyrrhetinic acid. The results obtained in validation assays indicate the accuracy and reliability of the developed simultaneous HPTLC method for the quantification of withaferin A and 18-β-Glycyrrhetinic acid in all three Ayurvedic formulations containing Ashwagandha and Glycyrrhiza.