Abstract: Flavonoids are the main bioactive components in propolis, which exhibit a variety of physiological activities such as anti-oxidative and antibacterial effects. Optimization of ultrasonic extraction of propolis flavonoids will provide a theoretical basis for exploitation and utilization of propolis products. Firstly, single-factor experiments are carried out. The effects of three extraction parameters (ultrasonic power, extraction time and alcohol concentration) on extraction yield of propolis flavonoids are investigated. Secondly, based on the single-factor tests, the ultrasonic extraction conditions of flavonoids from propolis are optimized by using response surface methodology. The experimental results show that the order of factors affecting the extraction yield is extraction time, alcohol concentration and ultrasonic power. The maximum extraction yield of propolis is 21.10%±0.075% (n=3) while ultrasonic power is 150 W, extraction time is 16 min and alcohol concentration is 86%. According to the analysis of this experiment, it is found that the propolis from Xiaogan, Hubei is identified as premium grade product in propolis. Compared with other ultrasonic extraction methods of propolis flavonoids, the ultrasonic extraction method used in this study has the advantages of lower extraction temperature and shorter extraction time.