To investigate the difference of biochemical indicators between decapitated blood, abdominal aortic blood and abdominal venous blood on in Wistar rat, and to provide the basis and reference of blood sampling method for toxicology tests.Methods 40 male and 40 female Wistar rat were divided into decapitated blood group and abdominal aortic/venous blood group. Serum parameters of TP, ALB, A/G, ALT, AST, BUN, CRE, CHO, TG, GLU were detected by automatic biochemical analyzer.Results All parameters were differences between these two blood sampling methods in two kinds of gender. The broken blood group in TP, ALB, A/G(female P<0.05), ALT, AST, CRE(female P<0.05), CHO, TG exceed the abdominal aortic blood group obviously on two sex(P<0.05), lower glucose in the broken blood group(P<0.05), BUN in the broken blood group exceed abdominal aortic blood group on female (P<0.05). Influences of aortic blood and venous blood embedded in serum parameters of TG, GLU, TG in the abdominal aortic blood group was lower on two sex(P<0.05), GLU in the abdominal aortic blood group exceed the abdominal venous blood group obviously on male(P<0.05), no difference in female.Conclusion There are significant influences on biochemistry parameters between different blood sampling methods and arterial blood and venous blood in Wistar rat.