Fluorescence detection of enzymatically formed hydrogen peroxide in aqueous solution and in reversed micelles

• Published in: Analytical biochemistry Vol. 187; no. 1; pp. 129 - 132
• Main Authors: Ferrer, Alvaro Sanchez, Santema, Jillert S., Hilhorst, Riet, Visser, Antonie J.W.G.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 15.05.1990; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Biochemie; Biochemistry; Biological and medical sciences; Cetrimonium; Cetrimonium Compounds; Colloids; Enzymes and enzyme inhibitors; Fluoresceins - analysis; Fluoresceins - metabolism; fluorescence; Fundamental and applied biological sciences. Psychology; Glucose - metabolism; Glucose Oxidase - metabolism; Hydrogen Peroxide - analysis; Methods; Micelles; Oxidation-Reduction; Oxidoreductases; Solutions; Spectrometry, Fluorescence; Glucose oxidase; Enzymatic activity; Enzyme; Reversed micelle; Aqueous solution; Detection; Fluorescence spectrometry; Hydrogen peroxides
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/0003-2697(90)90429-D

A sensitive enzymatic assay for oxidase reactions both in aqueous solution and in hexadecyltrimethylam-moniumbromide (CTAB) reversed micelles has been developed. The assay is based on the fluorescence detection of dichlorofluorescein, which is formed by hydrogen peroxide oxidation of the nonfluorescent precursor dichlorofluorescin. Hydrogen peroxide as product of the reaction catalyzed by glucose oxidase served to select the reaction conditions. The reaction rate is distinctly enhanced in CTAB reversed micelles as compared to the rate in aqueous solution. This effect, combined with the high sensitivity owing to the strong fluorescence of dichlorofluorescein, makes the assay attractive for the detection of low enzyme, substrate, or peroxide concentrations.