Preparative separation and purification of sulforaphene from radish seeds by high-speed countercurrent chromatography.

Abstract Sulforaphene, a kind of isothiocyanates, derived from glucoraphenin which is the important ingredient of radish ( Raphanus sativus L.) seeds, has shown significant pharmacological activities. In this paper, the separation and purification of sulforaphene from radish seeds, was achieved by high-speed countercurrent chromatography (HSCCC). A two-phase solvent system consisted of n -hexane–ethyl acetate–methanol–water (35:100:35:100, v/v/v/v) was applied. The revolution speed of the separation column, flow rate of the mobile phase and separation temperature were 800 rpm, 2 ml/min and 30 °C, respectively. From about 1000 mg amount of the crude plant extract, 249.4 mg of pure sulforaphene was obtained by one-step separation on a 280 ml HSCCC column. The purified sulforaphene was at a high purity of 96.9% and the mass recovery was more than 95%. The purity of sulforaphene was determined by HPLC analysis and its chemical structure was assessed by MS, 1 H NMR, 13 C NMR and DEPT-135 NMR.