title: Fluorescence detection of enzymatically formed hydrogen peroxide in aqueous solution and in reversed micelles.
abstract: A sensitive enzymatic assay for oxidase reactions both in aqueous solution and in hexadecyltrimethylammoniumbromide (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.
authors.affiliation: Department of Biochemistry, Agricultural University, Wageningen, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: Sanchez Ferrer
authors.forename: J S
authors.initials: JS
authors.lastname: Santema
authors.forename: R
authors.initials: R
authors.lastname: Hilhorst
authors.forename: A J
authors.initials: AJ
authors.lastname: Visser
mesh_terms: D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D003102:Colloids; D005452:Fluoresceins / Q000032:analysis* / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D005949:Glucose Oxidase / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000032:analysis*; D008722:Methods; D008823:Micelles; D010084:Oxidation-Reduction; D012996:Solutions; D013050:Spectrometry, Fluorescence
chemical_list: D002593:Cetrimonium Compounds; D003102:Colloids; D005452:Fluoresceins; D008823:Micelles; D012996:Solutions; C029569:diacetyldichlorofluorescein; C037631:2',7'-dichlorofluorescein; D006861:Hydrogen Peroxide; D005949:Glucose Oxidase; D005947:Glucose; D000077286:Cetrimonium