ORAC (Oxygen Radical Absorbance Capacity) Assay
Reexamination of the ORAC assay: effect of metal ions.
Nkhili E, Brat P.
Anal Bioanal Chem. 2011 May;400(5):1451-8
The oxygen radical absorbance capacity (ORAC) assay method has been employed extensively in the field of antioxidant and oxidative stress. It uses fluorescein as probe for oxidation by peroxyl radical. Hundreds of reports have been published on the use of this method to determine antioxidant capacity in food and biological samples. The question is whether the results of all these reports are influenced by antioxidant autoxidation, which occurs during the ORAC test. Indeed, the presence of metal ions in the studied matrix will influence antioxidant stability, thereby leading to the underestimation of their antioxidant properties. Ethylenediaminetetraacetic acid hydrate (EDTA) can be used as a metal complexation agent. This paper examines the effect of the addition of EDTA on the ORAC values of pure compounds (quercetin, ascorbic, and dehydroascorbic acid) and five food juices (kiwi, orange, tomato, red grape, and apple). Metal complexation by EDTA (80 μM) clearly increased the ORAC values, given that the antioxidant was protected against rapid autoxidation incited by trace metal ions within samples and then by free radicals. Our finding also undoubtedly demonstrated that the number of literature values is potentially underestimated. PMID: 21512895
Methods for the determination of antioxidant capacity in food and raw materials.
Litescu SC, Eremia S, Radu GL.
Adv Exp Med Biol. 2011;698:241-9.
A comprehensive description of the most frequently used methods to determine the antioxidant activity in food and raw materials is given. The methods are classified into two categories, depending on the type of the assessment carried out. Several methods for the assessment of antioxidant efficacy using free radical scavenging such as Oxygen Radical Absorbance Capacity Assay (ORAC), Total Radical Trapping Antioxidant Parameter assay (TEAC), Ferric reducing antioxidant power assay (FRAP) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) assay are described. An example of methods based on the assessment of antioxidant efficacy using significant biological substrates is also presented. Critical opinions concerning the proposed methods are presented.
PMID: 21706439