Corticosterone is one of the adrenal hormones. This paper reported the determination of corticosterone in plasma and tissue of small mammals with spectrofluorometry. The linear relationship between the fluorescence intensity and the concentration of corticosterone is over the range of 0.01-0.24 microg/mL (lambdaex = 470nm, lambdaem = 525nm). The average rate of standard recovery is 96.0% and RSD (n = 10) is 4.2%. This method is simple, rapid and sensitive.