Quantitative Analysis of Glycyrrhizic Acid in Crude Drug and its Herbal Formulation by HPLC

A rapid and accurate high performance reverse phase liquid chromatography has been developed for the quantitative determination of glycyrrhizic acid in crude drug and its herbal formulation. The developed method was validated in terms of s linearity, precision, accuracy, specificity, robustness, limit of detection and limit of quantitation according to International Conference on Harmonization guidelines and the statistical results obtained were within the acceptance criteria. Chromatography was carried out on BDS C-18 column (4.6 mm×250 mm, 5 μm) using isocratic mobile phase consisting of phosphate buffer and acetonitrile in the ratio of 45:55 (v/v) at a flow rate of 1 mL/min and eluents are monitored at 256 nm. The retention time of glycyrrhizic acid was found to be 2.26 minutes. The linear regression analysis data for the calibration plot showed a good linear relationship with correlation coefficient of 0.999 in the concentration range of 20 to 120 μg/mL for glycyrrhizic acid with respect to peak area. The limit of detection and limit of quantitation values were found to be 0.704 μg/ml and 2.348 μg/ml respectively. The % recovery value for glycyrrhizic acid is 99.67% confirms that the method is accurate. Due to its simplicity, rapidness and high precision, the proposed HPLC method may be used for the reliable quantitative determination of glycyrrhizic acid in both crude drug and herbal formulation.