Abstract: Objective This study investigates the clinical value of measuring total bile acid (TBA) and cholinesterase (CHE) simultaneously in the diagnosis, identification and prognosis of hepatitis B patients.Methods 256 hepatitis B patients were included in the study.TBA was measured by enzymatic cycling assay, cholinesterase was determined by rate method, and total bilirubin was determined by 2, 4-Dichloroaniline diazonium chromatometry.The results were compared with healthy controls.Results TBA of hepatitis B patients was significantly higher than the healthy controls (P<0.01) .The CHE level was significantly lower in moderate chronic hepatitis, severe hepatitis and primary liver cancer groups compared to healthy control group (P<0.01) .With exacerbations of chronic hepatitis, TBA and TBil levels gradually increased, and CHE was gradually decreased most significantly in the severe hepatitis B group (P<0.01) .TBA was positively correlated with TBil and negatively correlated with CHE.Conclusion Simultaneous determination of TBA and CHE is very important in the diagnosis, identification and prognosis of Hepatitis B patients.