Abstract: This study was designed to investigate experimental conditions for high-speed countercurrent chromatography (HSCCC) separation of the ethanol extract of Armillaria mellea fermentation broth supernatant. The best separation results were achieved using a solvent system composed of butanol, ethyl acetate and deionized water (1:4:5, V/V). Four peaks were separated well with an extraction yield of 51.10%, 1.47%, 0.86% and 1.80%, respectively.

Key words: Armillaria mellea fermentation broth supernatant, high-speed countercurrent chromatography, solvent system