Taking the glutinous rice flour as raw material，the response surface analysis methodology was used to optimize the alkaline method separation of glutinous rice starch. The optimum separation conditions were determined by Box-Behnken response surface methodology based on single factor test with the purity of glutinous rice starch as evaluation index. The results showed that the optimal separation conditions were room temperature, lye concentration 0.34%，liquid-material ratio 5:1and soaking time 4 h, under the conditions the purity of starch was up to 96.61%，with protein content less than 0.5%. The properties of glutinous rice starch were determined by scanning electron microscopy and rapid viscosity analyzer.