A peroxyoxalate chemiluminescence-based assay for the evaluation of hydrogen peroxide scavenging activity employing 9,10-diphenylanthracene as the fluorophore
2002; Elsevier BV; Volume: 48; Issue: 3 Linguagem: Inglês
10.1016/s1056-8719(03)00055-8
ISSN1873-488X
AutoresAnis Arnous, Christos Petrakis, Dimitris P. Makris, Panagiotis Kefalas,
Tópico(s)Antioxidant Activity and Oxidative Stress
ResumoIntroduction: A simple, rapid, sensitive, and enzyme-free analytical method for estimating scavenging of hydrogen peroxide (H2O2) was developed. Methods: Peroxyoxalate chemiluminescence (POCL) was measured, using 9,10-diphenylanthracene as fluorophore. Results: The chemiluminescence signal was found to be linear in response to increasing amounts of H2O2 in ethyl acetate/acetonitrile (9:1) (r2=.9990), within a range of concentrations varying from 9.0 to 72.0 μM. In contrast, acetonitrile was highly unsuitable because of poor linearity (r2=.3736) and poor signal stability. The linearity of POCL inhibition, as a measure of H2O2 scavenging, was tested employing well-known, lipid-soluble antioxidants, including β-carotene, butylated hydroxytoluene (BHT) and α-tocopherol, and also the more polar flavonol quercetin, and the water-soluble l-ascorbic acid (AA). Under the experimental conditions, the corresponding values of H2O2 scavenging activity (SAHP) for quercetin, β-carotene, α-tocopherol, and l-AA were 19.1±0.4, 70.9±20.1, 8.4±0.4, and 44.8±5.6×10−3 μM−1. Discussion: The data establish the assay as a method for assessing the H2O2 quenching activity of lipid-soluble antioxidants.
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