Reversed-phase high-performance thin-layer chromatographic method for the simultaneous determination of glycyrrhizin and glabridin biomarkers in glycyrrhiza glabra roots, rhizomes and herbal formulations

A new rapid, simple, economical, and environment-friendly reversed- phase high-performance thin-layer chromatography (RPHPTLC) method has been established for the simultaneous determination of glycyrrhizin and glabridin in Glycyrrhiza glabra roots, rhizomes and selected herbal formulations. The method was carried out using RP-18 silica gel 60 F254S HPTLC glass plates and methanol-water (7:3 v/v) as the mobile phase. The developed plates were scanned and quantified densitometrically at 256 and 233 nm for glycyrrhizin and glabridin, respectively. Glycyrrhizin and glabridin peaks from G. glabra roots and rhizomes and herbal formulations were identified by comparing their single spots at RF = 0.63 ± 0.02 and RF = 0.28 ± 0.01, respectively. Linear regression analysis revealed a good linear relationship between the peak areas and the amounts of glycyrrhizin and glabridin in the ranges of 1000-7000 and 100-700 ng band-1, respectively. The method was validated, in accordance with the International Conference on Harmonization (ICH) guidelines for precision, accuracy, and robustness. The proposed method will be useful to determine the therapeutic doses of glycyrrhizin and glabridin in herbal formulations as well as in bulk drug.