The coupling reaction of 4-aminoantipyrine (4-AAP) with phenol using the superoxide anion radical (O2-*) as oxidizing agent under the catalysis of horseradish peroxidase (HRP) was studied. Based on the reaction, O2-* produced by irradiating vitamin B2 (VB2) was spectrophotometrically determined at 510 nm. Under the optimum experimental conditions, the relationship between A510 and O2-* concentration was linear in the range 9.14x10(-6)-1.2x10(-4) mol L(-1). The detection limit was determined to be 1.37x10(-6) mol L(-1). A possible reaction mechanism was discussed. The effect of interferences and surfactants on the determination of O2-* was also investigated. The proposed method was applied to determine superoxide dismutase activity in garlic, scallion, and onion with satisfactory results.