High-speed counter-current chromatography (HSCCC) was used to separate and purify two isoflavones for the first time from () mycelium using a two-phase solvent system composed of chloroform-dichloromethane-methanol-water (4:2:3:2, ///). These two isoflavones were identified as genistein (4',5,7-trihydroxyisoflavone, CHO₅) and daidzein (4',7-dihydroxyisoflavone, CHO₄), using infrared spectroscopy (IR), electro-spary ionisation mass (ESI-MS), ¹H-nuclear magnetic resonance (NMR) and C-NMR spectra. About 23 mg genistein with 95.7% purity and 18 mg daidzein with 97.3% purity were isolated from 150 mg ethanolic extract of mycelium. The results demonstrated that HSCCC was a feasible method to separate and purify genistein and daidzein from mycelium.
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