Abstract: The oil was prepared by screw press. The sample was methyl-esterificated and saponificated, and then was analyzed by GC-MS. The aging model of mice made by D-galactose was established, and then was fed with tobacco seed oil. The level of MDA in the blood, the levels of GSH and protein carbonylation and the activities of GSH-Px in the liver were measured. The results showed that the tobacco seed oil contained 2 saturated fatty acids, 3 unsaturated fatty acids, 8 sterols and squalene. UFA in the oil was as high as 88.53%, total sterol content was 12.14mg/g and squalene content was 0.24 mg/g. The tobacco seed oil significantly decreased MDA content in the blood and protein carbonyl content in the liver, and increased GSH content and GSH-Px activities in the liver. Our conclusion is that tobacco seed oil could be a very good raw material of edible oil, and has significant antioxidant activities.