Supercritical carbon dioxide (SC-CO2) extraction of whole fruit oil from Gardenia jasminoides Ellis was performed. The effect of extraction pressure, temperature and CO2 flow rate on the oil yield was investigated by response surface methodology (RSM). The results showed that experimental data had a good fit to the proposed model (R 2 = 0.938). Extraction pressure, CO2 flow rate, the quadratics of pressure, and the interaction between pressure and flow rate showed significant effects on the oil yield (p < 0.05). The optimum parameters that maximized the yield of gardenia fruit oil (GFO) were: extraction pressure of 36.8 MPa, temperature of 65 °C, and CO2 flow rate of 15 kg/h. The main fatty acid of GFO was linoleic acid (about 44%), followed by palmitic acid (about 26.4%) and oleic acid (about 24.6%). α-Tocopherol was dominant in the total tocopherols of GFO, and showed the main antioxidant activity. The fatty acid composition and tocopherols content of GFO were not remarkably affected by the extraction by SC-CO2 and n-hexane.