This work studied the extraction of perilla oil from perilla seeds using supercritical carbon dioxide, which is valued for its high omega-3 content. Response surface methodology was used to investigate the optimum conditions of perilla oil extraction at pressures of 200, 225, and 250 bar and temperatures of 40 degrees C, 50 degrees C, and 60 degrees C. The fatty acid composition was also investigated. The optimal condition was obtained at 60 degrees C under a pressure of 218 bar. Under this extraction condition, the predicted measures of oil yield (%), acid value, peroxide value, iodine value, saponification value, total phenolic content, DPPH (IC50), ABTS (IC50), and tocopherol were: 37.00%, 1.22 mg KOH/g, 0.94 mEq O2/kg, 205.00 g I2/100 g, 196.90 mg KOH/g, 12.14 mg GAE/100 g, 10.93 mg/mL, 154.86 mg/mL, and 604.58 mg/kg, respectively. In addition, gas chromatography analysis indicated that perilla seed oil was primarily composed of linolenic acid (56.94-58.02%), with a total polyunsaturated fatty acid content of 75.49-76.47%, while saturated fatty acids and monounsaturated fatty acids constituted 10.53-11.11% and 13.00-13.52% of the total fatty acids, respectively.