Objective To analyze the metabolic characteristics of myocardial infarction (MI) using metabolomics to better understand its pathogenesis and to explore new therapeutic directions for MI. Methods Serum metabolites in ten acute MI mice and five sham-control mice were analyzed by UHPLC-QqQ/MS, and SPSS was used for statistical analysis. MetaboAnalyst 5.0 was used to analyze the metabolic pathways of the differential metabolites and build a metabolic network. Results One hundred and twenty-nine metabolites were detected by UHPLC-QqQ/MS. Significant serum metabolite differences were found between MI mice and normal controls. Fifty out of 129 metabolites in serum were associated with MI. In addition, the most important metabolic pathways were D-glutamate metabolism, alanine, aspartate and glutamate metabolism, glycine, serine and threonine metabolism, glyoxylate and dicarboxylate acid metabolism. ConclusionMetabolites in serine-related metabolic pathways reduce in serum in MI. We propose a new therapeutic direction for myocardial protection in MI.