Ferric 2-hydroxy-1-naphthaldehyde thiosemicarbazone (Fe I I I -HNT) has been synthesized and used to mimic the active group of horseradish peroxidase (HRP). The catalytic characteristics of this mimic-enzyme catalyst in the oxidation reaction of ascorbic acid with hydrogen peroxide has been studied. The experimental results showed that this peroxidase model compound has similar catalytic activity that of HRP. The catalytic activity of Fe I I I -HNT has been compared with those of HRP and common mimic-enzymes, metalloporphyrins (e.g. metal tetrakis(sulphophenyl)porphyrin (Me-TPPS 4 )). Though the catalytic activity of Fe I I I -HNT is 75% of that of HRP, it can be used as a new kind of mimic-enzyme catalyst in determination of trace H 2 O 2 . By coupling this mimetic catalytic reaction with the catalytic reaction of glucose oxidase, glucose can be determined indirectly. Under experimental conditions the linear relationship between ΔA 2 6 5 and glucose concentration was in the range of 2.0-40.0 μg ml - 1 , The correlation coefficient (r) was 0.9996. The R.S.D. (P=0.05, n=6) was 0.24%. The detection limit was determined to be 0.238 μg ml - 1 . It was applied successfully to the determination of glucose in normal human and diabetic urine. The values of determination by the proposed method were compared with those of a routine method (enzymic glucose determination) applied in an hospital clinical laboratory. The agreement was very good.