Abstract: Objective: To investigate the voluntary blood donors serum alanine aminotransferase (ALT) reference range in Xuzhou area. Method: ALT values of 6677 blood donors were determined by rate method, and the results were statistically analyzed according to age and gender. Result: The ALT value of voluntary blood donors in Xuzhou area was 7.3~50.1 U/L, and the ALT value of male and female was 8.7~58.2 U/L and 6.6~38.5 U/L, respectively. There was a significant difference between male and female groups(P<0.01). The ALT value of 18 to 25 years old group, 26 to 35 years old group, 36 to 45 years old group and 46 to 55 years old group was 6.8~45.1 U/L, 7.9~59.9 U/L, 8.2~57.2 U/L and 8.8~48.2 U/L. There were significant differences between 18 to 25 age groups and other age groups(P<0.01). The ALT level showed a skewed distribution in age. Conclusion: The ALT reference range of blood donors in Xuzhou area serum was close to that of clinical commonly used enzyme reference interval issued by the Ministry of Health. There were significant differences between the male and female population, as well as the 18 to 25 years old age group and other age groups. Therefore it should make different reference ranges for different groups of people, to ensure the blood quality and safety and reduce blood discarding.