Abstract: In order to improve the extraction of total saponins from Nigella glandulifera Freyn et Sint,the extraction rate of total saponins of Nigella glandulifera Freyn et Sint was evaluated as an index. The effect of enzyme preparation,enzyme dosage,ethanol concentration,solid-liquid ratio,enzymatic hydrolysis time,ultrasonic frequency and enzymatic hydrolysis temperature were investigated by single factor experiment. The response surface methodology was used to optimize the best process for enzyme-assisted ultrasonic extraction of total saponins from Nigella glandulifera Freyn et Sint. Results showed that the best technology of enzyme-assisted ultrasonic extraction of total saponins from Nigella glandulifera Freyn et Sint was:The enzymolysis temperature 60℃,the ethanol concentration 54%,the ratio of solid to liquid 1:37 g/mL,the amount of enzyme 0.98%,the ultrasonic frequency 60 kHz,the time 30 min,the actual extraction rate was 14.09%±0.12%,and the model equation predicted value was 14.17%,relative error was less than 3%,which indicated that the optimal conditions were feasible.