Objective To compare the effects of two different samples and different test methods on blood biochemical indexes and blood glucose values in rats. Methods Glucose levels in the serum and plasma samples were detected with a blood glucometer, and the biochemical parameters in the serum and plasma were determined by routine blood biochemistry.The data were statistically compared and analyzed to determine if there are some significant differences.Results Among the 19 biochemical indexes of serum and plasma specimens, nine parameters, i.e., ALB,TP,ALP,CHOL,URIC,GLU,Mg,LD,Ca showed significant differences(P< 0.05),while the other 10 indexes showed no significant difference (P> 0.05).Different samples and different methods had significant differences in the detection of blood glucose (P< 0.05). Conclusions Different method and samples impact on the detection of blood glucose and some other serum and plasma biochemical indexes.