Abstract: In this study, the root of Toddalia asiatica (L.) Lam. was used as the experimental material and the content of total alkaloids was considered as the detection index. The optimum extraction conditions of total alkaloids in T. asiatica roots were investigated using response surface methodology (RSM). The optimum extraction conditions were: ethanol concentration of 74.76%, material-solvent ratio of 20∶1, and extraction time of 32.87 min, with a theoretical total alkaloid content of 11.09 mg/g. According to the optimum extraction conditions, the average value of three parallel experiments was (10.74 ±0.18) mg/g, which showed little difference from the theoretical content. Thus, the optimum extraction conditions of total alkaloids in T. asiatica obtained by the RSM were accurate and reliable, and this mathematical model was well fitted and of practical value.