ObjectiveTo investigate the reference range of enzymes in urine samples of normal rats.MethodsFour-hour urine samples were collected from 95 normal rats. Urinary enzymes including NAG, ALP, γ-GT, LDH were analyzed using a biochemical analyzer. Reference ranges were shown with upper 95th percentile. ResultsThe reference ranges for female rats:NAG<5.3 U/L,ALP<110.8 U/L,γ-GT<753.0 U/L,LDH<32.6 U/L; for male:NAG<27.3 U/L,ALP<329.0 U/L,γ-GT<769.8 U/L,LDH<40.0 U/L. Reference ranges using the unit/g·Cre for female:NAG<28.9 U/g,ALP<723.5 U/g,γ-GT<1763.2 U/g,LDH<113.1 U/g; for male rats:NAG<84.3 U/g,ALP<680.2 U/g,γ-GT<2522.3 U/g,LDH<121.8 U/g.ConclusionThe reference ranges of urinary enzymes may provide background values for renal toxicity evaluation in the future.