Abstract: The optimal extraction conditions of Grifola frondosa were studied. The effects of pH value, extraction time, solid-liquid ratio and extraction temperature on the extraction rate of polysaccharides from Grifola frondosa were studied by single factor test. On the basis of single factor test, four factors and three levels orthogonal experiments were set up to optimize the extraction process with the extraction rate of Grifola frondosa polysaccharides as the index. The results showed that the order of influencing factors was extraction time > pH value > solid-liquid ratio > extraction temperature. The optimum extraction condictions were pH 7.5, extraction time 2.0 h, solid-liquid ratio 1∶20,and extraction temperature 90℃. With these conditions, the extraction rate was 20.94%.