Objective: Using metabolomics methods to study the metabolic profiles of amino acids and lysophosphatidylcholine (LPC) in serum of rats with non-alcoholic fatty liver disease(NAFLD), and to identify biomarkers that can characterize NAFLD and speculate on the possible mechanism of its occurrence. Methods: NAFLD rats were prepared by feeding high-fat diet feed and intraperitoneal injection of carbon tetrachloride. The levels of 15 LPCs and 18 amino acids in the serum of the control and NAFLD groups were determined using liquid chromatography-mass spectrometry.SPrincipal component analysis and orthogonal partial least squares discriminant analysis were used to analyze the changes in serum LPC and amino acid metabolic profiles in NAFLD rats. Pearson correlation analysis was used to analyze the correlation between biomarkers and NAFLD.SResults: The metabolic profiles of serum LPC and amino acid in the NAFLD group significantly deviated from the control group and could be completely distinguished.SLPC (20:1), arginine and glutamic acid had significant contributions to NAFLD, and these species were identified as biomarkers. Furthermore, LPC (20:1) and arginine were significantly correlated with serum biochemical indicators such as AST, ALT, LDL, and TBIL.SConclusions: The metabolic profiles of serum LPC and amino acid were closely related to NALFD.