This study investigated a novel, simple, and economical method for the preparation and purification of glabridin from licorice roots. Glabridin was initially obtained from ethyl acetate extraction of licorice, followed by using solid phase extraction (SPE) and preparative high performance liquid chromatography (HPLC). The content of glabridin increased from 0.23% to 35.2% after SPE, and then a 16mg product at a high purity of over 95% was obtained from 10g licorice roots after purification by preparative HPLC. The purity was assessed by analytical HPLC, and the purified compound was characterized by LC-MS/MS and 1H NMR.