Fluorometric Determination of Hydrogen Peroxide in Natural Water Samples by Flow Injection Analysis with a Reaction Column of Peroxidase Immobilized onto Chitosan Beads

Abstract

It was found that a FIA system with an immobilized enzyme reaction column could be utilized for the determinations of hydrogen peroxide in some water samples, such as rain water and tap water. A column was packed with chitosan beads immobilizing horseradish peroxidase. Under the recommended conditions, it took 5 min for each assay, and the detection limit, which was defined as a signal-to-noise ratio of 3, was 3.0 ng dm −3 of hydrogen peroxide. The relative standard deviation at 1.0 mg dm −3 of hydrogen peroxide was 1.5% (n=20). There was no interference from some coexisting substances having a low oxidation-reduction ability or not; therefore, a standard addition method could be applied to rain water and tap water samples. From such results, it has become clear that the proposed FIA system can be applied to the determination of hydrogen peroxide in water samples, and that the system is suitable for routine analysis in environmental observations.

Monitoring of Hydrogen Peroxide, Nitrate and Nitrite in Rain Water

“…Moreover, there had been developed a flow-injection system for the determination of hydrogen peroxide with an immobilized horseradish peroxidase reaction column; it was applied for the determination of hydrogen peroxide in rain water. 39 From the results obtained by this system, it was found that mg dm -3 levels of hydrogen peroxide were often present in rain water. In this paper, we describe the relationship between hydrogen peroxide and nitrate and nitrite from monitoring of these substances in rain water.…”

“…Immobilized horseradish peroxidase (EC1.11.1.7) was prepared as described in a previous paper. 39 An immobilized enzyme column was made using a plastic mini-column (5 mm i.d. × 50 mm) packed with about 0.8 g of it.…”

“…39 An aliquot of the sample solution (0.3 cm 3 ) was injected into the carrier stream through an injection valve (Rheodyne 7125) and mixed with an HPPA solution as a fluorometric indicator. The mixture reacted in the immobilized horseradish peroxidase column to produce a fluorescent species.…”

Fluorometric Determination of Glucose by Flow Injection Analysis Using Immobilized Enzyme-Reactor Columns Prepared by Coupling Glucose Oxidase and Peroxidase onto Chitosan Beads

“…It was observed that the peak height decreased if Tris buffer was excluded in the carrier solution, as was the case in our previous paper. 11 Therefore, Tris buffer was set at 0.05 mol dm -3 in the carrier solution with reference to previous paper. However, the effect of the concentration of Tris buffer and the pH of the carrier did not influence the sensitivity.…”

“…[5][6][7][8][9] Therefore, an immobilized horseradish peroxidase (POD-IE) was prepared by a glutaraldehyde bridging method using chitosan beads as a carrier; 10,11 it was possible to apply it to the determination of hydrogen peroxide by flow injection analysis (FIA) using 3-(p-hydroxyphenyl)propionic acid (HPPA) as a fluorescent substrate and a fluorometric detector.…”