Abstract: Hemoglobin was used as a mimetic peroxidase in the catalytic oxidation reaction of p-cresol by hydrogen peroxide. The hemoglobin-catalyzed reaction was coupled with the oxidation of glucose catalyzed by GOD to determine glucose by fluorimetric method. Under optimum conditions, the calibration graph for the determination of glucose was linear over range of 0.0～5.0×10^-5mol/L glucose, with detection limit of 6.5×10^-8mol/L glucose. The method was used in determination of glucose of human sera with satisfactory results.