To investigate the antioxidant activity of recombinant human lactoferrin (rhLF). Methods The 1,1-diphenyl-2-picryl hydrazyl free radical (DPPH) inhibiting rate, hydroxyl free radical inhibiting rate, superoxide anion radical inhibiting rate, reducing capacity and inhibiting of lipid peroxidation in rat liver were tested to evaluate the antioxidant activity of rhLF and bovine lactoferrin (bLF) in vitro. Results The concentration of rhLF and bLF were correlated with the inhibiting rate of 1,1-diphenyl-2-picryl hydrazyl free radical, hydroxyl free radical, superoxide anion radical and reducing capacity. There was significantly dose-response relationship and the difference had statistical significance(P＜0.05). There was no interaction between sample and concentration, the trend of the value was the same in different concentrations of rhLF and bLF. Antioxidant effect of rhLF and bLF had significant difference in rat liver homogenate, different concentrations of rhLF and bLF had significant difference(P＜0.05). Conclusion Bovine lactoferrin and rhLF have antioxidant activity, and could be added to health food or cosmetics as a natural antioxidant.