Superoxide dismutase (SOD) activity was measured using a flow injection-chemiluminescent assay (FI-CLA) based on xanthinexanthine oxidase dependent superoxide (O2.) formation. The mobile phase consists of 50 mM potassium phosphate buffer (pH 7.4) containing lucigenin (5 µM) and xanthine (0.3 mM). Under optimum conditions, bovine albumin did not affect chemiluminescence at concentrations of 11000 µg mL1 and KCN inhibited 100% of the Cu, Zn-SOD activity using 5 µL of a 0.23 mM concentration. The analysis of one sample was done in less than 30 s with a relative standard deviation of ±3.1%. SOD activity in the biological samples was correlated to the amount of exogenously applied SOD. The FI-CLA method reported here appears to be one of the faster and more useful tools used to assay SOD activity.Key words: superoxide dismutase, flow injection-chemiluminescent assay, superoxide, xanthine oxidase, lucigenin.