HPTLC Method for the Estimation of Glycyrrhizaglabra in PatoladiGhrita- An Ayurvedic Polyherbal Formulation

Abstract

The quantitative estimation of crude drugs present in an Ayurvedic formulation still remains a big challenge. A quantitative High Performance Thin Layer Chromatography (HPTLC) method was developed for the estimation of Yasti(Glycyrrhizaglabra), one of the herbal ingredients present in the ayurvedicpolyherbal formulation ‘PatoladiGhrita’. The formulation ‘PatoladiGhrita’ is a medicated ghee originally described in the ayurvedic book ‘Ashtangahridaya’ and is given in the Ayurvedic Formulary of India.The methanol extracts of the crude drug Yasti and the formulation PatoladiGhritawere run using a mobile phase, Chloroform : Methanol : Acetic acid 7:2:1, on a silica G 60 F₂₅₄ stationary phase. Methanol extract of Yastigave a spot with R_f value of 0.51. The methanol extract of PatoladiGhritaalso gave a spot with R_f value of 0.51. The chemical equivalency of these two spots was revealed by the same pattern of the UV spectrum. The estimation was done graphically by preparing ‘Quarter Standard’, ‘Half Standard’, ‘Normal Standard’, and ‘Double Standard’ of the formulation. The methanol extract of the genuine drug and that of the four standards and four marketed samples were run using the same mobile phase. The reference spots on each track were scanned and integrated to get the Area Under the Curve (AUC).A calibration plot was drawn between the concentration of standards and AUC of the reference spots. The concentration of Yasti in the samples was found from the corresponding AUC obtained for the reference peak of the samples. A regression coefficient of 0.988 was obtained. The developed HPTLC method is a simple and accurate one and can be used as a tool for the quantification of each ingredient in a polyherbal formulation.