Abstract: The bovine blood was used as raw material to extract hemoglobin by means of ultrasonic. Effect of ratio (V/V) of water to red blood cell, operation time, ratio of ultrasonic time to quiescent interval and ultrasonic power on the extraction ration of hemoglobin were investigated. Through single factor test, their scopes were initially determined, and then the extraction technology parameters were optimized through response surface methodology. The results showed that :when the ratio (V/V) of water to red blood cell was 4∶1, the operation time was 17 min, the ratio of ultrasonic time to quiescent interval was 1 ∶1, and the ultrasonic power was 250 W, the extraction rate of hemoglobin was 92.33%.