A Simple Semi‐preparative Reversed‐phase HPLC/PDA Method for Separation and Quantification of Glycyrrhizin in Nine Samples of <i>Glycyrrhiza glabra</i> Root Collected from Different Geographical Origins
Abstract
INTRODUCTION: Glycyrrhiza glabra L. (Fabaceae), commonly known as 'liquorice', is one of the most popular ingredients in several traditional herbal medicinal preparations, and glycyrrhizin is the major glycoside present in this plant. The content of glycyrrhizin may vary among G. glabra samples collected from various geographical origins, which may affect the therapeutic efficacy. Thus, quantification of glycyrrhizin in G. glabra samples is important. OBJECTIVE: To develop and validate a simple semi-preparative reversed-phase HPLC with photodiode array (PDA) method for separation and quantification of glycyrrhizin in nine samples of G. glabra root collected from various geographical origins. METHODS: Dried and ground root of G. glabra was Soxhlet-extracted sequentially with n-hexane and methanol (MeOH). The separation and quantification of glycyrrhizin was achieved on a C18 reversed-phase semi-preparative column using a gradient mobile phase, 30-100% solvent B in solvent A in 30 min (solvent A: 0.1% v/v trifluoroacetic acid (TFA) in water and solvent B: 0.1% v/v of TFA in MeOH), at a flow rate of 3.00 mL/min and UV detection at 254 nm. RESULTS: A simple semi-preparative reversed-phase HPLC/PDA method allowing clear separation and quantification of glycyrrhizin content in nine samples has been validated in terms of linearity, selectivity, limits of detection, precision, accuracy and detection. Concentration levels of glycyrrhizin were between 0.177 and 0.688% w/w of dry materials. CONCLUSION: This method is precise, less time consuming and more cost effective, and can be used for the quality control of any G. glabra sample with regard to its glycyrrhizin contents.