This study was carried out to investigate the emulsifying properties of surface-active substances from defatted rapeseed cake by supercritical CO₂extraction. Based on the interfacial tension data, a supercritical fluid extract (SFE) with the lowest value of 14.16 mN/m was chosen for evaluation which was obtained from No. 2 extraction condition (150 bar, 65℃, 250 g). For emulsions with SFE, some physicochemical properties (i.e., fat globule size, creaming stability, zeta potential etc) were investigated according to changes in SFE concentration, pH, and NaCl addition in an emulsion. It was found that fat globule size was decreased with increasing SFE concentration in emulsion, with showing a critical value at 0.5 wt%, thereby resulting in less susceptibility to creaming behavior. The SFE emulsion also showed instability at acidic conditions (pH<7.0) as well as by NaCl addition. This was coincided with zeta potential data of emulsion. In addition, SSL (sodium stearoyl lactylate) found to be suitable as a co-surfactant, as it helped considerably in decreasing fat globule size in emulsions and its optimum concentration to be over 0.03 wt%, based on 0.1 wt% SFE in emulsion.