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

Phenolics are broadly distributed in the plant kingdom and are the most abundant secondary metabolites of plants. Plant polyphenols have drawn increasing attention due to their potent antioxidant properties and their marked effects in the prevention of various oxidative stress associated diseases such as cancer. In the last few years, the identification and development of phenolic compounds or extracts from different plants has become a major area of health- and medical-related research. This review provides an updated and comprehensive overview on phenolic extraction, purification, analysis and quantification as well as their antioxidant properties. Furthermore, the anticancer effects of phenolics in-vitro and in-vivo animal models are viewed, including recent human intervention studies. Finally, possible mechanisms of action involving antioxidant and pro-oxidant activity as well as interference with cellular functions are discussed.

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Plant phenolics: extraction, analysis and their antioxidant and anticancer properties.

Processed fruits and vegetables have been long considered to have lower nutritional value than their fresh commodities due to the loss of vitamin C during processing This research group found vitamin C in apples contributed < 04% of total antioxidant activity, indicating most of the activity comes from the natural combination of phytochemicals This suggests that processed fruits and vegetables may retain their antioxidant activity despite the loss of vitamin C Here it is shown that thermal processing elevated total antioxidant activity and bioaccessible lycopene content in tomatoes and produced no significant changes in the total phenolics and total flavonoids content, although loss of vitamin C was observed The raw tomato had 076 +/- 003 micromol of vitamin C/g of tomato After 2, 15, and 30 min of heating at 88 degrees C, the vitamin C content significantly dropped to 068 +/- 002, 064 +/- 001, and 054 +/- 002 micromol of vitamin C/g of tomato, respectively (p < 001) The raw tomato had 201 +/- 004 mg of trans-lycopene/g of tomato After 2, 15, and 30 min of heating at 88 degrees C, the trans-lycopene content had increased to 311+/- 004, 545 +/- 002, and 532 +/- 005 mg of trans-lycopene/g of tomato (p < 001) The antioxidant activity of raw tomatoes was 413 +/- 036 micromol of vitamin C equiv/g of tomato With heat treatment at 88 degrees C for 2, 15, and 30 min, the total antioxidant activity significantly increased to 529 +/- 026, 553 +/- 024, and 670 +/- 025 micromol of vitamin C equiv/g of tomato, respectively (p < 001) There were no significant changes in either total phenolics or total flavonoids These findings indicate thermal processing enhanced the nutritional value of tomatoes by increasing the bioaccessible lycopene content and total antioxidant activity and are against the notion that processed fruits and vegetables have lower nutritional value than fresh produce This information may have a significant impact on consumers' food selection by increasing their consumption of fruits and vegetables to reduce the risks of chronic diseases

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Thermal Processing Enhances the Nutritional Value of Tomatoes by Increasing Total Antioxidant Activity

Non-structural phenolic compounds perform a variety of functions in plants, including acting as antioxidants. We describe a microplate-adapted colorimetric total phenolics assay that utilizes Folin-Ciocalteu (F-C) reagent. The F-C assay relies on the transfer of electrons in alkaline medium from phenolic compounds to phosphomolybdic/phosphotungstic acid complexes, which are determined spectroscopically at 765 nm. Although the electron transfer reaction is not specific for phenolic compounds, the extraction procedure eliminates approximately 85% of ascorbic acid and other potentially interfering compounds. This assay is performed in microcentrifuge tubes and assessed in a 96-well plate reader. At least 64 samples can be processed in 1 d.

Estimation of total phenolic content and other oxidation substrates in plant tissues using Folin-Ciocalteu reagent.

Determination of the total phenolic, flavonoid and proline contents in Burkina Fasan honey, as well as their radical scavenging activity.

Publisher Summary This chapter discusses the analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Analyses of the Folin-Ciocalteu (FC) type are convenient, simple, and require only common equipment and have produced a large body of comparable data. Under proper conditions, the assay is inclusive of monophenols and gives predictable reactions with the types of phenols found in nature. Because different phenols react to different degrees, expression of the results as a single number—such as milligrams per liter gallic acid equivalence—is necessarily arbitrary. Because the reaction is independent, quantitative, and predictable, analysis of a mixture of phenols can be recalculated in terms of any other standard. The assay measures all compounds readily oxidizable under the reaction conditions and its very inclusiveness allows certain substances to also react that are either not phenols or seldom thought of as phenols (e.g., proteins). Judicious use of the assay—with consideration of potential interferences in particular samples and prior study if necessary—can lead to very informative results. Aggregate analysis of this type is an important supplement to and often more informative than reems of data difficult to summarize from various techniques, such as high-performance liquid chromatography (HPLC) that separate a large number of individual compounds .The predictable reaction of components in a mixture makes it possible to determine a single reactant by other means and to calculate its contribution to the total FC phenol content. Relative insensitivity of the FC analysis to many adsorbents and precipitants makes differential assay—before and after several different treatments—informative.

Vernon L. Singleton

Papers

Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent