Abstract: Objective: The Non-targeted metabolomics technology was used to analyze the metabolites in the serum and liver of mice with chronic alcoholic liver injury (ALD), to identify and screen out the differential metabolites related to alcoholic liver injury and construct its metabolic pathway, and to explore the protective mechanism of puerarin against alcoholic liver injury in mice. Methods: The mouse model of chronic alcoholic liver injury was established by intragastric administration of 52 liquor for 8 weeks. Meanwhile, the protective effects of puerarin on ALD mice were evaluated using biochemical indexes of liver function and pathological tissue sections. Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) metabonomics technique was used to analyze the metabolites of mouse liver and serum. Multivariate statistics were used to process the data, and the variable importance projection (VIP) of the first principal component was >1 and the t-test P<0.05 was used to screen out the differential metabolites. Metabolic pathways of differential metabolites using KEGG database. Results: Puerarin could significantly improve ALT and AST levels in alcohol-induced liver injury mice (P<0.05 or P<0.01). Furthermore, the pathological section results indicated that puerarin could improve liver inflammatory cell infiltration in ALD mice. Through multivariate statistics, 11 metabolites were found in serum metabolites between the model group and the blank group. Under puerarin intervention, the levels of 11 metabolites tended to blank group, mainly related to amino acid synthesis and metabolism, glutathione metabolism, porphyrin metabolism and chlorophyll metabolism. There were 32 different metabolites between the model group and blank group in liver metabonomics, and the levels of 31 different metabolites tended to blank group under puerarin intervention, which mainly involved the amino acid synthesis and metabolism, biosynthesis of primary bile acids, purine metabolism, phosphatidylinositol signalling system, steroid metabolism and other metabolic pathways. Conclusion: Puerarin has a protective effect on alcoholic liver injury in mice, and its mechanism may be through regulating the disorder of serum and liver metabolites, which provides a valuable reference for anti-alcoholism health food in the future.