Abstract: This study aimed to assess the safety and toxicity of peony protein and provide a scientific basis for its application in food industry. The safety evaluation included acute oral toxicity tests in mice, as well as 28 d and 90 d oral toxicity tests in rats. The results demonstrated that no signs of poisoning or mortality were observed in the acute oral toxicity test (intragastric dose of 11700 mg/kg) conducted on mice. Furthermore, in the 28 d and 90 d oral toxicity tests conducted on rats, no abnormal changes were observed in terms of food utilization, organ weight ratios, and various physiological activities, when compared to the control group (1500, 3000, 6000 mg/kg). Additionally, there were no significant differences (P>0.05) in blood routine and blood biochemical indices between the treatment groups and the control group. Histopathological examination of organ tissues revealed intact morphology without any toxic lesions. These findings provided solid evidence that peony protein was non-toxic in the dose range of less than 11700 mg/kg and hold promising potential for application in the food industry.