The fatty acid composition, volatile components, rheological properties and antioxidant activity of yak butter at different altitudes (2 500~2 800 m, 3 500~4 000 m and above 4 300 m) in Tibetan were analyzed and compared. The results showed that altitude had significant effects on fatty acid composition, volatile components, rheological properties, and antioxidant activity (P<0.05). The unsaturated fatty acid content of yak butter at altitude of 2 500~2 800 m was the highest. Yak butter with an altitude of more than 4 300 m had pure smell and best flavor quality. Yak butter at altitudes of 3 500~4 000 m had the best fluidity. Yak butter at different altitudes had different antioxidant abilities, among which yak butter at altitudes ranging from 2 500 m to 2 800 m had the strongest DPPH free radical (66.99 mmol/100 g Trolox), hydroxyl free Trolox (26.71 mmol/100 g Trolox), superoxide anion scavenging ability (22.85%), and iron ion chelating antioxidant ability (1.41%).