Objective: To investigate the anti-fatigue effect of L-arginine complex preparation on mice. Methods: The experimental mice were divided into a blank group, low dose group, medium dose group and high dose group. L-arginine complex preparation mice were intragastrically administered for 30 days, and the mice were tested for exhaustive swimming time. At the same time, contents of plasma lactic acid, lactate dehydrogenase, urea nitrogen and hepatic glycogen were measured. Results: Compared with the blank control group, the weight-bearing swimming time and hypoxia-tolerant survival time of the low, middle and high dose groups was significantly increased (P<0.05). Whereas, the serum urea nitrogen levels and lactose content were significantly decreased (P<0.05). However, compared with the blank control group, the liver glycogen content of the middle and high dose groups was increased significantly (P<0.05), and there was no significant difference in the low dose group. Conclusion: The L-arginine complex preparation has an anti-fatigue function in mice.