Isolation and Purification of Two Isoflavones from Hericium erinaceum Mycelium by High-Speed Counter-Current Chromatography

Abstract

High-speed counter-current chromatography (HSCCC) was used to separate and purify two isoflavones for the first time from Hericium erinaceum (H. erinaceum) mycelium using a two-phase solvent system composed of chloroform-dichloromethane-methanol-water (4:2:3:2, v/v/v/v). These two isoflavones were identified as genistein (4',5,7-trihydroxyisoflavone, C₁₅H₁₀O₅) and daidzein (4',7-dihydroxyisoflavone, C₁₅H₁₀O₄), using infrared spectroscopy (IR), electro-spary ionisation mass (ESI-MS), ¹H-nuclear magnetic resonance (NMR) and ¹³C-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 H. erinaceum mycelium. The results demonstrated that HSCCC was a feasible method to separate and purify genistein and daidzein from H. erinaceum mycelium.

Keywords: Hericium erinaceuns mycelium; daidzein; genistein; high-speed counter-current chromatography (HSCCC).

Figure 1

HPLC chromatograms of H. erinaceum mycelium crude extract (HEM-E-E). Peak number 1 and 2 refer to compounds 1 and 2.

Figure 2

HSCCC chromatogram of HEM-E-E under the optimized condition. The upper phase of chloroform-dichloromethane-methanol-water (4:2:3:2, v/v/v/v) system was used as the stationary phase and lower phase of these solvent system was used as mobile phase. HSCCC condition was as follows: flow rate 2.0 mL/min, column temperature 20 °C, sample loading 10 mL, sample content 150 mg/10 mL, detection wavelength = 254 nm, rotary speed= 900 rpm. Peak number 1 and 2 refer to compounds 1 and 2.

Figure 3

HPLC chromatograms of compound 1 (A) and compound 2 (B) and UV wavelength scanning of compounds 1 and 2 (inside). Peak number 1 and 2 refer to compounds 1 and 2.

Figure 4

Chemical structures of genistein and daidzein.