Abstract: Orthogonal design was used to optimize the extraction process of rubusoside from Rubus suabissimus S.Lee. On the basis of single-factor test, the factors ( solid-liquid ratio, ethanol concentration, ultrasonic power, p H) and levels were selected.According to the orthogonal design principle and analysis method, the optimum conditions were 2.6% of complex enzyme ( cellulase∶ pectinase and∶ papain = 1∶ 1∶ 1) concentration, 4.0 of p H value, 1∶ 20 of solid-liquid ratio, 40% of concentration of ethanol, 50 W of ultrasonic power, 40 of mesh sieves, 30 min of enzyme hydrolysis, 45 ℃ of extract temperature, 30 min of ultrasonic extraction time, 2 times of repeated extraction.Under the optimal extraction conditions, the rate of rubusoside was66.4% ± 0.9% and the content was 110.7 mg/g.