Flow Injection Analysis of Hydrogen Peroxide Using Glass-Beads Modified with Manganese(III)-Tetra(4-carboxyphenyl)porphine Derivative and Its Analytical Application to the Determination of Serum Glucose

Abstract

A flow injection analysis system of hydrogen peroxide was developed. The present system is based on measuring of the absorbance of a quinoid dye formed by the following reaction catalyzed by peroxidase: Phenol + 4-Aminoantipyrine + Peroxidase $2{{H}_{2}}{{O}_{2}}\xrightarrow{Peroxidase}$ Quinoid dye + 4H2O. A column packed with aminopropyl-glass beads modified with a manganese(III)-tetra(4-carboxyphenyl)porphine derivative (Mn-TCPPG column), which has peroxidase-like activity, was used in place of an immobilized peroxidase column in the above reaction. The linear range of the calibration curve was 0.4–80 μg/ml hydrogen peroxide. The relative standard deviation of this system was 2.97% (n = 100, 10 μg/ml hydrogen peroxide 20 μl injection). The Mn-TCPPG column has sufficiently stability for the continuous injection of hydrogen peroxide untill 100 times. The advantageous feature of the Mn-TCPPG column was a less-electrostatic interaction between the mother glass beads and the anionic chromogen or quinoid dye formed and the stability in terms of the storage, temperature and moisture. The determination of serum glucose was achieved by attaching an immobilized glucose oxidase column to this system without deproteinization.