Abstract: Antroquinonol, a metabolite produced from solid-state fermentation of Antrodia camphorata has been shown great anti-cancer effect. This study investigated the effects of a number of factors including fermented grain types, substrate capacity, initial moisture content (containing the water of seeds) , supplementary carbon source (glucose) , nitrogen content on Antroquinonol content by Antrodia cinnamomea and the multi-stage countercurrent extraction method for extracting Antroquinonol from the solid-state fermentation products of Antrodia camphorata in order to improve the production of Antroquinonol in solid-state fermentation, the extraction efficiency, decrease loss and provide theoretical basis for preparation of Antroquinonol at industry scale. The results were as follows:the best compositions of fermentation medium for Antroquinonol content were as follows:rice 110g, initial moisture content 50% (with inoculated seed liquid water) , carbon source 2.0g (glucose) , nitrogen content 0. 3g (soybean powder) . With these optimum conditions, Antroquinonol content increased by 14.44% than the initial value;After three-stage countercurrent extraction by 95% ethanol (volume ratio of ethanol and fermentation product was 10∶1, v/w) in 50°C water bath and shaking for 1h, the results showed that the residual rate of Antroquinonol in the fermentation products was less than 1%. This method showed the less solvent used and higher extraction efficiency.