Abstract: In order to extract total flavonoids from silkworm pupa of sericin variety effectively and quickly, we used the method of ultrasonic-assisted technique to conduct the experiments. Through single factor experiments, we focused the impact of the four factors of total flavonoids extracted, including ethanol concentration, solid-liquid ratio, extraction time, extraction temperature, to determine the best single factors. The optimal extraction conditions were determined by orthogonal experiments. The results showed that the optimum extraction process of extracting total flavonoids from silkworm pupa of sericin variety by ultrasonic-assisted technique was that the ultrasonic power 500 W, the ethanol concentration 40%, liquid ratio 1：40 (W/V), extraction temperature 60℃, extraction time 40 min. Under these conditions, the extraction rate was up to 2.017%.