Background and aims Bile acids, which modulate the glycose and lipid metabolism, play an important role in the development of non-alcoholic fatty liver disease (NAFLD). This study aims to explore the association between serum total bile acid (TBA) and non-alcoholic steatohepatitis (NASH) in the general population from a large clinical dataset. Methods NAFLD individuals confirmed by ultrasonography were enrolled in the study. The NASH was defined as the NAFLD individual with abnormal liver function, and non-alcoholic fatty liver (NAFL) was defined as the NAFLD individual with normal liver function. The demographic characters, biochemical results including serum TBA were compared between NASH and NAFL patients. The independently associated factors with NASH were investigated by multivariate logistic regression analysis. Results A total of 6862 individuals were enrolled in the study, of which 23.7% were NASH. The median age was 45 (39-55) years old, and the BMI was 26.9(25.1-28.9) kg/m 2 . The NASH patients showed higher levels of serum TBA compared to the NAFL patients [3.30(2.30-5.0) μmol/L vs. 2.70(1.90-4.20) μmol/L, P <0.001]. The prevalence of NASH tended to increase with the increasing of serum TBA level ( P <0.001). Multivariate logistic regression analysis showed the age, male, serum TBA levels, diabetes mellitus (DM), serum uric acid, total cholesterol and triglyceride were independent risk factors for NASH ( P <0.05). Stratification analysis according to DM status showed that serum TBA was independently associated with NASH in individuals without DM (OR: 1.55, 95% CI: 1.11-2.13, P =0.008), while not in individuals with DM (OR: 1.32, 95% CI: 0.53-2.96, P =0.515). Conclusion Serum TBA level was significantly higher in NASH patients than in NAFL patients, and was independently associated with NASH in non-diabetes population.