Abstract: To study the antioxidant activity and protective effect on CCl₄ liver injuried mice of total flavonoids from genuine traditional chinese drug Isodon amethystoides flower with different polarities, and to provide the foundation for effective utilization reasonably of the resources of I. amethystoides and exploit the natural antioxidants, the ethanol crude extraction was obtained by extracting the total flavonoids assisted by ultrasound, then the petroleum ether phase, ethyl acetate phase, n-butanol phase and water phase were extracted successively. The concentration of total flavonoids in each phase and its scavenging effects on DPPH,·OH, O₂^-·and the reducing power on FRAP were measured. Acute liver injuried model of mice induced by CCl₄. The activity of ALT and AST in serum, and the T-AOC, SOD activity, MDA concentration in serum and liver homogenate from mice which was administrated by total flavonoids with different polarities were evaluated. The results showed that the content of total flavonoids in the ethyl acetate phase was highest, and it had the best scavenging effect on DPPH,·OH, O₂^-·and FRAP reducing powder. The activity of ALT and AST in mice serum from each extract phase group was significantly higher than the model group, and the T-AOC, SOD activity in mice serum and liver homogenate from ethyl acetate phase group was also significantly higher than the model group. The MDA content was significantly lower than the model group. The investigation results showed that the total flavonoids from Isodon amethystoides flower with different polarities had significant antioxidant activity and protective effect on CCl₄ liver injuried mice.