Surface-active substances in defatted rapeseed cake were obtained using a supercritical fluid extraction method. Then, it was purified by removing sinapine in the extract through a series of steps using a mixed solvent: diethyl ether and ethyl acetate (1:1, v/v). Emulsifying properties of purified surface-active substances were investigated, including fat globule size, zeta potentials and creaming stability and its antioxidant activity in emulsion systems were also studied by peroxide value and 1H-NMR spectrum. It was found that fat globules in emulsions with purified surface-active substances were much smaller than ones with the unpurified. In addition, as pH of the emulsion lowered and with increasing NaCl concentration in the emulsion, they were observed to increase, which led to worse creaming stability. These properties were reflected in changes of zeta potentials of emulsions. The oxidative stability was better in emulsions with purified surface-active substances than ones with Tween 20 or commercial lecithin, possibly resulted from the existence of sinapic acid in the extract. It was concluded that purified surface-active substances from defatted rapeseed cake could be simultaneously used as emulsifier and antioxidant agent in emulsion system.