Preparative Isolation and Purification of Glyceollins from Soy Bean Elicited with Aspergillus sojae by High-Speed Countercurrent Chromatography

Abstract

High-speed countercurrent chromatography (HSCCC), a support free liquid-liquid partition chromatographic technique, has been used for the preparative isolation of isoflavonoid glyceollins from the ethanol extracts of elicited soy bean seeds. The preparative HSCCC instrument is a multilayer coil planet centrifuge equipped with three preparative coils, separation was performed with a two phase solvent n-hexane/EtOAc/MeOH/H2O (3:4:2:1, v/v/v/v) at a flow rate of 1.0 mL/min in ‘head-to-tail’ mode. HSCCC separation yielded 6.0 mg of glyceollins and with purities > 86.90 %. Chemical structures of glyceollin compounds were elucidated by ESI– MS, UPLC–MS.