Objective To observe the effects of different sample process modes on blood biochemical indicator detection in Wistar rats. Methods 40 Wistar rats of specific pathogen free(SPF)level, weighing 180~220 g, were took with half males and half females. After 12 h fasting, rats were anaesthetized by 3% pentobarbital sodium, blood was collected and divided into two samples. One of the samples was for whole blood and another for anticoagulation, with 3mL per serving. Each blood sample was put into three 1.5 mL centrifuge tubes and three heparin anticoagulative tubes with 1mL per, and centrifuged after putting 1 h, 2 h and 4 h in room temperature for testing blood glucose, blood lipid, protein, myocardial enzymes, indexes of liver and renal function. Results The types of blood samples and the standing time after collection and before centrifugation had no significant effect on TC, TG, ALB, ALT, ALP, BUN and CREA in serum or in plasma of Wistar rats(P>0.05).But the value of GLU in serum was significantly higher than that in plasma (P<0.05, P<0.01). With the extension of storage time after collection and before centrifugation, the value of GLU continued to decline. The values of AST, CK, LDH in serum were significantly higher than that in plasma (P<0.05, P<0.01). With the extension of storage time after collection and before centrifugation, the value of AST, CK, LDH continued to increase. However, there was no significant effect on the values of AST, CK, LDH by placing different time after collection and before centrifugation(P>0.05). Conclusion Types of blood samples had no significant effect on blood lipid, protein and indexes of renal function. However, detection value of GLU, AST, CK and LDH would be influenced on a certain degree. The extension of standing time after collection and before centrifugation resulted in plasma GLU decreasing and plasma AST, CK and LDH increasing, but has no obvious effect on the serum AST, CK and LDH.