Abstract: Response surface methodology was used to optimize the extraction conditions of cyclopamine from Veratrum grandiflorum. Based on single factor experiments, the liquid-to-material ratio extraction time, extraction temperature were chosen as causal factors and the extraction of cyclopamine was chosen as response value which was then analyzed by models. In this way, the three-factors-three-levels Box-Behnken central composition experiments were designed. Results showed that liquid-to-material ratio, extraction time, extraction temperature had significant effects (p<0.01) on extraction of cyclopamine. Optimal conditions for achieving the high extraction of cyclopamine were liquid-to-material ratio 9∶1 (mL/mg) , extraction time 10.0h, extraction temperature 48.0℃. Under these optimized conditions, with 10.0g raw materials extraction get cyclopamine 9.38mg in the abstract, the actual extraction of cyclopamine was 9.19mg, and the model prediction was worth 2.02 percent error.