title: Dietary antioxidants interfere with Amplex Red-coupled-fluorescence assays.
abstract: Oxidation of Amplex Red by hydrogen peroxide in the presence of horseradish peroxidase (HRP) gives rise to an intensely colour product, resorufin. This reaction has been frequently employed for measurements based on enzyme-coupled reactions that detect hydrogen peroxide as a final reaction product. In the current study, we show that the presence of dietary antioxidants at biological concentrations in the reaction medium produced interferences in the Amplex Red/HRP catalyzed reaction that result in an over quantification of the hydrogen peroxide produced. The interference observed showed a dose-dependent manner, and a possible mechanism of interaction of dietary antioxidants with HRP in the Amplex Red-coupled-fluorescent assay is proposed.
authors.affiliation: Department of Experimental Medicine, Faculty of Medicine, University of Lleida-IRBLleida, Lleida, Spain. jceserrano@mex.udl.cat
authors.forename: José
authors.initials: J
authors.lastname: Serrano
authors.forename: Mariona
authors.initials: M
authors.lastname: Jové
authors.forename: Jordi
authors.initials: J
authors.lastname: Boada
authors.forename: María Josep
authors.initials: MJ
authors.lastname: Bellmunt
authors.forename: Reinald
authors.initials: R
authors.lastname: Pamplona
authors.forename: Manuel
authors.initials: M
authors.lastname: Portero-Otín
mesh_terms: D000975:Antioxidants / Q000737:chemistry*; D004032:Diet; D036681:Fluorescence Recovery After Photobleaching / Q000592:standards*; D006735:Horseradish Peroxidase / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000032:analysis* / Q000737:chemistry; D010078:Oxazines / Q000737:chemistry*; D010084:Oxidation-Reduction
chemical_list: D000975:Antioxidants; D010078:Oxazines; C470430:Amplex Red; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase