Antioxidants and Radical Scavengers

Abstract: Food antioxidants play an important role in the food industry due to their ability

to neutralise free radicals that might be generated in the body. They do that by donating

their own electrons to free radicals without becoming free radicals in the process themselves,

hence terminating the radical chain reaction. The converted free radical products will then

be eliminated from the body before causing any harm; in this regard, antioxidants play the

role of scavengers protecting body cells and tissues. In this chapter, the processes which lead

to the formation of these reactive species (free radicals) and the different additives used as

antioxidants or radical scavengers to counter the effects of free radicals will be discussed.

Sources of different types of antioxidants, the various mechanisms by which they work and

analytical methods for determination and quality control are also examined.

Keywords: antioxidants; free radical species; ORAC assay; HORAC assay; DPPH assay;

FRAP assay; Trolox; TEAC assay; ABTS assay; PCL assay; DMPD assay; DL assay;

TBARS assay; Brigg-Rauscher assay

1.1 CHEMISTRY OF FREE RADICALS AND ANTIOXIDANTS

1.1.1 Introduction

From the viewpoint of chemistry, free radicals refer to any molecule with an odd unpaired

electron in its outer electronic shell, a configuration responsible for the highly reactive nature

of such species. The presence of such highly reactive free radicals in biological systems is

directly linked to the oxidative damage that results in severe physiological problems. The

free radical species that are of concern in living systems include the reactive oxygen species

(ROS), superoxide radicals (SOR), hydroxyl radicals and the reactive nitrogen species (RNS).

The oxygen-containing reactive species are the most commonly occurring free radicals in

living medium and are therefore of greatest concern. The oxidative damage caused by these

free radicals can be prevented by using antioxidants which include enzymatic antioxidant

systems such as catalase, glutathione peroxidase and superoxide dismutase (SOD) as well

as non-enzymatic antioxidants (Figure 1.1). It should be noted that, in nature, the generation
of free radicals which cause oxidative stress and that of antioxidants or radical scavengers is

carefully controlled such that there is always a balance between the two (Vouldoukis et al.

2004). Examples of non-enzymatic antioxidants include vitamin C (ascorbic acid) which

is a sugar acid, vitamin E (-tocopherol) and -carotene, bilirubin, propyl gallate (PG, a

condensation ester product of gallic acid and propanol), uric acid, tertiary butylhydroquinone
(t-BHQ), butylated hydroxyanisole (BHA), ubiquinone and macromolecules which include
ceruloplasmin, albumin and ferritin. Generally, mixtures of different antioxidants provide
better protection against attack by free radicals rather than individual antioxidants.

Due to the importance of antioxidant systems, there are a number of quality assess-
ment criteria for the antioxidant performance of these systems. Various assays have been

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Antioxidants and Radical Scavengers 3

developed to assess the antioxidant capacities, including the oxygen radical absorbance

capacity (ORAC) assay, ferric reducing ability of plasma (FRAP), Trolox equivalent antiox-
idant capacity (TEAC) assay, etc. Antioxidant foods which are dietary nutrients containing

antioxidant compounds and non-nutrient antioxidants which are normally added to foods to
play the role of antioxidants will be discussed simultaneously in this chapter, unless indicated