Abstract: Objective To investigate the effect of different methods of irrigating the amputated limbs of rats with the University of Wisconsin solution (UW) to find out which method has the strongest effect on preserving and prolonging the vitality of amputated limbs. Methods The lower extremities were amputated from 30 healthy adult male Wistar rats and randomly divided into 3 groups (10 rats in each group). Rats in the three groups were irrigated with UW solution from femoral artery and the amputated limbs were preserved in refrigerator at 4℃. In group A, limbs were irrigated continuously, and they were irrigated intermittently for 20 minutes every 4 hours in group B, for 1 hour every 12 hours in group C with the speed of 12 ml/h. The irrigated outflow was collected to test the biochemical indexes and the samples of skeletal muscle were taken for pathological and biochemical examination regularly after the amputation. Results There were no significant differences in alkaline phosphatase, alanine aminotransferase and glucose at different time points between group A and group B, C with similar pathological change. No significant difference was found in pathological and ultrastructural change, biochemical indexes between group B and group C. Conclusion The effect of preserving amputated limbs in 4℃ with 4 h intermittently irrigating and 12 h intermittently irrigating on protecting skeletal muscle is similar with continuously irrigating. And there is no significant difference between 4 h intermittently irrigating and 12 h intermittently irrigating.