Objective To screen potential biomarkers to explore the pathogenesis in a rat model of isoproterenol- induced cardiac hypertrophy. Methods Isoproterenol 30 mg / ( kg·d) was used to establish the rat model of cardiac hypertrophy via intraperitoneal injection for 14 consecutive days. The rat cardiac hypertrophy model was evaluated via the cardiac index. Ultra high performance liquid chromatography-quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF- MS) was conducted to detect serum metabolites in normal and model rats. MPP software was used to analyze metabolic differences. The Human Metabolome Database (HMDB) was used to identify biomarkers. MetaboAnalyst 4. 0 was used to analyze metabolic pathways. Results Serum metabolites in the rats with isoproterenol-induced cardiac hypertrophy differed significantly from those of the normal rats, and 10 potential biomarkers were identified. Compared with the normal group, sphingosine 1-phosphate and dihomo-γ-linolenic acid in the model group were significantly downregulated, and D-fructose- 1,6-bisphosphate, deoxyadenosine, N-acetylmethionine, phytosphingosine, allantoin, 3-keto-β-D-galactose, octane, and glycerol were significantly upregulated. Conclusions The metabolic pathways involved in isoproterenol-induced cardiac hypertrophy include sphingolipid metabolism, glycerolipid metabolism, galactose metabolism, biosynthesis of unsaturated fatty acids, and purine metabolism. This study provides a basis for understanding the metabolic changes in the circulating blood in a model of isoproterenol-induced cardiac hypertrophy.