Extraction and separation of glabridin from licorice by reversed phase high performance liquid chromatography

The extraction and separation of glabridin from licorice root by HPLC was performed in this work. First, by investigating the different extraction solvents, extraction methods and extraction times, a one-hour ultrasonic extraction procedure with ethyl acetate as the extraction solvent was optimized. Then the ethyl acetate extraction was applied to RP-HPLC for separation of glabridin. The column efficiencies and resolutions were experimentally investigated with different mobile phase compositions. Baseline separation of glabridin was obtained under the mobile phase composition of 50/50 vol.% (ACN/water). The retention time of glabridin was 20.3 min. The peak of glabridin was collected from the HPLC elution for several times and identified by LC/MS. Under the optimum extraction and HPLC separation methods, 1.26 g of glabridin per kg licorice root could be extracted.