Abstract: The extraction process of estradiol from milk was investigated in this paper. Based on single factor experiments,response surface methodology was used to explore the effects of liquid-material ratio,microwave power and microwave time on extraction yield of estradiol. The results showed that the optimal conditions for the extraction of estradiol were the liquid-material ratio of 4.1∶1,microwave power of 475 W,microwave time of61 s. Under the optimal extraction conditions,the highest extraction rate of estradiol was up to 92.77%. Compared with the ultrasonic extraction method,microwave extraction method had short extraction time,high extraction yield,low energy consumption,environmental protection and other advantages.