This review summarizes the multifaceted aspects of antioxidants and the basic kinetic models of inhibited autoxidation and analyzes the chemical principles of antioxidant capacity assays. Depending upon the reactions involved, these assays can roughly be classified into two types:  assays based on hydrogen atom transfer (HAT) reactions and assays based on electron transfer (ET). The majority of HAT-based assays apply a competitive reaction scheme, in which antioxidant and substrate compete for thermally generated peroxyl radicals through the decomposition of azo compounds. These assays include inhibition of induced low-density lipoprotein autoxidation, oxygen radical absorbance capacity (ORAC), total radical trapping antioxidant parameter (TRAP), and crocin bleaching assays. ET-based assays measure the capacity of an antioxidant in the reduction of an oxidant, which changes color when reduced. The degree of color change is correlated with the sample's antioxidant concentrations. ET-based assays include th...

The Chemistry behind Antioxidant Capacity Assays

Methods available for the measurement of antioxidant capacity are reviewed, presenting the general chemistry underlying the assays, the types of molecules detected, and the most important advantages and shortcomings of each method. This overview provides a basis and rationale for developing standardized antioxidant capacity methods for the food, nutraceutical, and dietary supplement industries. From evaluation of data presented at the First International Congress on Antioxidant Methods in 2004 and in the literature, as well as consideration of potential end uses of antioxidants, it is proposed that procedures and applications for three assays be considered for standardization: the oxygen radical absorbance capacity (ORAC) assay, the Folin-Ciocalteu method, and possibly the Trolox equivalent antioxidant capacity (TEAC) assay. ORAC represent a hydrogen atom transfer (HAT) reaction mechanism, which is most relevant to human biology. The Folin-Ciocalteu method is an electron transfer (ET) based assay and gives reducing capacity, which has normally been expressed as phenolic contents. The TEAC assay represents a second ET-based method. Other assays may need to be considered in the future as more is learned about some of the other radical sources and their importance to human biology.

https://faculty.missouri.edu/~glaserr/3700s14/Antioxidant-Capacity_jf0502698.pdf

Standardized Methods for the Determination of Antioxidant Capacity and Phenolics in Foods and Dietary Supplements

There has been increasing interest in the research on flavonoids from plant sources because of their versatile health benefits reported in various epidemiological studies. Since flavonoids are directly associated with human dietary ingredients and health, there is need to evaluate structure and function relationship. The bioavailability, metabolism, and biological activity of flavonoids depend upon the configuration, total number of hydroxyl groups, and substitution of functional groups about their nuclear structure. Fruits and vegetables are the main dietary sources of flavonoids for humans, along with tea and wine. Most recent researches have focused on the health aspects of flavonoids for humans. Many flavonoids are shown to have antioxidative activity, free radical scavenging capacity, coronary heart disease prevention, hepatoprotective, anti-inflammatory, and anticancer activities, while some flavonoids exhibit potential antiviral activities. In plant systems, flavonoids help in combating oxidative stress and act as growth regulators. For pharmaceutical purposes cost-effective bulk production of different types of flavonoids has been made possible with the help of microbial biotechnology. This review highlights the structural features of flavonoids, their beneficial roles in human health, and significance in plants as well as their microbial production.

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Chemistry and Biological Activities of Flavonoids: An Overview

Publisher Summary This chapter discusses ferric reducing/antioxidant power (FRAP) assay. The ferric reducing/antioxidant power (FRAP) assay is a recently developed, direct test of “total antioxidant power.” The FRAP assay is robust, sensitive, simple, and speedy and facilitates experimental and clinical studies investigating the relationship among antioxidant status, dietary habits, and risk of disease. Measurement of the total antioxidant power of fresh biological fluids—such as blood plasma—can be measured directly; the antioxidant content of various dietary agents can be measured objectively and reproducibly and their potential for improving the antioxidant status of the body investigated and compared. The FRAP assay is also sensitive and analytically precise enough to be used in assessing the bioavailability of antioxidants in dietary agents to help monitor longitudinal changes in antioxidant status associated with an increased intake of dietary antioxidants and to investigate the effects of disease on antioxidant status.

Ferric reducing/antioxidant power assay: direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration

https://aggie-horticulture.tamu.edu/faculty/cisneros/Papers/Thaipong_2006.pdf

Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimating antioxidant activity from guava fruit extracts

http://www.radio.cuci.udg.mx/bch/EN/Manuals/Techniques/FRAP-original_AnalytBiochem_1996-v239-p70.pdf

The ferric reducing ability of plasma (FRAP) as a measure of 'antioxidant power' : the FRAP assay

This study aimed to compare in vitro antioxidant power of different types of tea (Camellia sinensis). The ferric reducing/antioxidant power (FRAP) assay was used to measure the total antioxidant power of freshly prepared infusions of 25 types of teas. Results showed that different teas had widely different in vitro antioxidant power and that the antioxidant capacity was strongly correlated (r = 0. 956) with the total phenolics content of the tea. Expressed as micromol of antioxidant power/g of dried tea leaves, values ranged as 132-654 micromol/g for black ("fermented") teas, 233-532 micromol/g for Oolong ("semifermented") teas, and 272-1144 micromol/g for green ("nonfermented") teas. One cup of tea of usual strength (1-2%), therefore, can provide the same potential for improving antioxidant status as around 150 mg of pure ascorbic acid (vitamin C).

Total antioxidant capacity of teas by the ferric reducing/antioxidant power assay.

Epidemiological evidence links high intake of ascorbic acid (AA) and other antioxidant micronutrients to health promotion. It would be useful to know the overall, or 'total' antioxidant capacity of foods, to establish the contribution of AA to this, and to assess how this information may translate into dietary intakes to meet the new US daily reference intake for AA. In this study, the total antioxidant capacity, as the ferric reducing-antioxidant power (FRAP) value, and AA content of thirty-four types of fruits and vegetables were measured using a modified version of the FRAP assay, known as FRASC. This measures AA (reduced form only) simultaneously with the FRAP value. Results covered a wide range: 880-15940 micromol/kg fresh wet weight and <20-540 mg/kg fresh wet weight respectively, for FRAP and AA, which comprised < 1-73 % and < 1-59 % total antioxidant capacity of fruits and vegetables respectively. We estimate that 100 mg AA is contained in one orange, a few strawberries, one kiwi fruit, 1-2 slices of pineapple, several florets of raw cauliflower or a handful of uncooked spinach leaves. Apples, bananas, pears and plums, the most commonly consumed fruits in the UK, contain very little AA. Results indicate also that the antioxidant capacity of vegetables decreases rapidly and significantly after fragmentation. Results of this, and future studies, using FRASC as a biomonitoring tool will be useful in food production, preparation, preservation, and aid dietary choices to increase antioxidant and AA intake. Furthermore, FRASC will facilitate bioavailability studies of antioxidants from different foods of known antioxidant capacity and AA content.

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Total antioxidant and ascorbic acid content of fresh fruits and vegetables: implications for dietary planning and food preservation.

In this review the process of lipid peroxidation and the atherogenicity of peroxidied lipids are discussed. Recent findings with regard to the effect of selected dietary factors on susceptibility of lipids to oxidative stress and on antioxidant defences are analysed with particular reference to their potential use in the prevention and treatment of atherogenesis and, by extension, coronary heart disease. Laboratory methods of assessing antioxidant defences, lipid peroxidation and the effects of lipid peroxidation are also reviewed and discussed with particular reference to their ability to assess in vivo oxidative stress and lipid peroxidation status. A range of oxidative stress indices are presented and their limitations discussed, but the main focus is on the most commonly used laboratory test for lipid peroxidation, the thiobarbituric acid reacting substances (TBARS) test. Finally, the influence of selected dietary factors on measured peroxidation status is discussed, with particular reference to the antioxidant vitamins C (ascorbic acid) and E (alpha tocopherol) and the type of fatty acids (mono- and poly-unsaturated) in the diet.

Iris F.F. Benzie

Papers

The ferric reducing ability of plasma (FRAP) as a measure of 'antioxidant power' : the FRAP assay

Ferric reducing/antioxidant power assay: direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration

Total antioxidant capacity of teas by the ferric reducing/antioxidant power assay.

Total antioxidant and ascorbic acid content of fresh fruits and vegetables: implications for dietary planning and food preservation.

Lipid peroxidation: A review of causes, consequences, measurement and dietary influences