A feasible methanol/n-hexane sequential extraction method for producing non-toxic oil from Jatropha curcas seeds is proposed with a high oil recovery. The method uses whole Jatropha curcas seeds as the plant material. The method utilizes response surface methodology to optimize the conditions of the first extraction step for phorbol esters. Phorbol esters enriched fraction is obtained to the maximum by this method. A rapid and sensitive detection method for phorbol esters content by ultra-performance liquid chromatography is proposed.