Abstract: Objective To analyze the level of alanine aminotransferase（ALT）, aspartate aminotransferase（AST）, alkaline phosphatase（ALP）, gamma glutamyltransferase（GGT） among healthy children and adolescents, and compare the differences between genders and ages. Methods A total of 2 351 healthy children and adolescents aged from 3 months to 18 years were divided into 4 groups: infant group, pre-school group, school age group and youth group. The serum enzymes in venous blood were detected by dry chemical system, and data were analyzed by statistical methods between different ages and genders. Results Except the ALP of school age group in girls was significantly higher than that in boys （P<0.05）, the AST, GGT of school age group and the ALT, AST, ALP, GGT of youth group in boys was significantly higher than that in girls （P<0.05）. The serum enzymes showed significant differences in different age groups. The trend of 4 enzymes increase or decline with increasing age （P<0.05）, but ALP showed no significant difference between infant and school age group, ALT showed no significant difference between pre-school and school age group （P>0.05）. We should set up different reference intervals according to the different age group. Conclusion The serum enzymes have gender differences and significantly associated with the change of age in healthy children and adolescents, and the laboratory shall establish a corresponding reference intervals.