Development of Analytical Methods (RP-HPLC and HPTLC) for the Fast Analysis of Glabridin in Crude Drug and Unani Formulations

Glabridin [4-[(3R)-8,8-Dimethyl-3,4-dihydro-2H-pyrano[6,5-f]chromen-3-yl]benzene-1,3-diol] is a major polyphenolic flavonoid specific for Glycyrrhiza glabra L., which has been reported to possess several pharmacological activities such as antioxidant, anti-Helicobacter pylori, antifungal, estrogenic, anti-proliferative and antinephritic activity [1, 2, 3]. There is a dearth of analytical methods using glabridin as analytical marker for quality control of Glycyrrhiza and formulations containing it, therefore, simple, economic, well validated, precise and an accurate HPLC and HPTLC methods were proposed for determination of glabridin for the quality control as an alternative to glycyrrhizin. HPLC chromatographic experiments were conducted on YL9100 HPLC system with UV detector using reversed-phase chromatography on a C18 column (250mm x 4.6µm) with water and acetonitrile as mobile phase in gradient elution method at a flow rate of 1.0 mL min^-1. HPTLC analysis was performed using toluene: dichloromethane: ethyl acetate in equal ratios. Methanol is used as solvent for preparation of sample and standard in both cases. Detection was done at 230 and 286nm for HPLC and HPTLC, respectively. Both HPLC and HPTLC methods were validated as per the ICH guidelines and the amount of glabridin present in crude as well as polyherbal formulation was quantified. The wide linearity range, sensitivity, accuracy and simple mobile phase imply that both the methods are suitable for routine quantification of glabridin with high precision and accuracy.

Acknowledgements: Authors are grateful to CCRUM-AYUSH, Ministry of Health and Family Welfare, Government of India, for providing financial assistance.

References: [1] Carmeli E, et al. (2008)J Basic Clin Physiol Pharmacol, 19: 49–63. [2] Fukai T, et al. (2002) Life Sci, 71: 1449–1463. [3] Fatima VK, et al. (2009) Phytother Res, 23: 1190–1193.