Antioxidants activity, quantification

An interesting development is the combination of HPLC and on-line measurement of reducing capacity or antioxidative activity. This approach allows both direct identification of antioxidative species in complex foods and quantification of the contribution to the overall antioxidative capacity in the absence of synergistic and antagonistic effects. Major advantages are less sample handling and the ability to rim large series of samples in an automated process. 

Netzel, M. et al., Sources of antioxidant activity in Australian native fruits. Identification and quantification of anthocyanins, J. Agric. Food Chem. 54, 9820, 2006. 

Soong YY and Barlow PJ. 2006. Quantification of gallic acid and ellagic acid from longan (Dimocarpus longan Lour.) seed and mango (Mangifera indica L.) kernel and their effects on antioxidant activity. Food Chem 87(3) 524-530. 

The products formed during lipid peroxidation include unsaturated aldehydes, such as 4-hydroxynonenal. Their quantification is of great interest because of their extremely reactive and cytotoxic properties. This extreme reactivity and metabolic conversion, however, may make them unsuitable as test analytes for in vivo antioxidant activity studies except at high levels of oxidative stress. Furthermore, simple chemical tests such as the TBARS (thiobarbituric acid reactive substances) and LPO-586 (colorimetric... 

The photochemiluminiscence (PCL) assay was initially used by Popov and others (1987). Popov and Lewin (1994 1996) have extensively studied this technique to determine water-soluble and lipid-soluble antioxidants. The PCL assay measures the antioxidant capacity, toward the 02 radical, in lipidic and water phase. This method allows the quantification of both the antioxidant capacity of hydrophilic and/or lipophilic substances, either as pure compounds or complex matrices from different origin synthetic, vegetable, animal, human, etc. The PCL method is based on an approximately 1,000-fold acceleration of the oxidative reactions in vitro by the presence of an appropriate photosensitizer. The PCL is a very quick and sensitive method. Chua and others (2008) used this assay to determine the antioxidant potential of Cin-namomum osmophloeum, whereas Kaneh and Wang and others (2006) determined the antioxidant capacity of marigold flowers. The antioxidant activity of tree nut oil extracts was also assessed by this method (Miraliakbari and Shahidi 2008). 

Hermann (2000) described a rapid automated method involving generation of a known amount of free radicals and the detection of the excess by photochemiluminescence. Test kits are available for determination of total water-soluble antioxidants, fat-soluble antioxidants and ascorbic acid. A luminometric method was developed for the determination of antioxidative activity and was subsequently applied to anthocyanin and betalaine colour concentrates (Kuchta et al., 1999). The method involved quantification of the interruption in luminescence from the hydrogen peroxide-horse radish peroxidase-luminol system in the presence of antioxidants. 

Thus, antioxidant activity is a parameter that permits quantification of the capacity of a compound (natural or artificial) and/or a biological sample (from a wide range of sources) to scavenge free radicals in a specific reaction medium. ... 

Antioxidant activity can be measured in a number of different ways. The most commonly used methods are those in which a chromogenic radical compound is used to simulate ROS and RNS it is the presence of antioxidants that provokes the disappearance of these chromogenic radicals, as shown in the reaction model given in Scheme 1. In order for this method to be effective, it is necessary to obtain synthetic metastable radicals that can easily be detected by photometric or fluorimetric techniques. Nevertheless, different strategies for the quantification of antioxidant activity have been utilized e.g., decoloration or inhibition assays. Details of these strategies and commonly used methods have been presented and reviewed elsewhere. 

Determinations of antioxidant activity are widely used in phytochemistry, nutrition, food chemistry, clinical chemistry, as well as in human, animal, and plant physiology, etc. Methods adapted to HPLC have appeared only recently but can be expected to have multiple apphcations in the future. ARTS" is an excellent metastable chromogen for the detection and quantification of the HAA and LAA of biological samples. Thus, using a simple photometer (end-point method), a microplate reader (multisample titration method), or HPLC equipment, a broad range of possibilities are available for the characterization of diverse samples (animal- or plant-derived). Some apphcations of special interest could include ... 

Extensive reviews of analytical methods for anthocyanins (Francis, 1982 Jackman et al., 1987b Strack and Wray, 1994) and other flavonoids (Williams and Harbome, 1994) as well as phenolic acids (Herrmann, 1989) have been published. In these reviews, extraction procedures, methods for fractionation of groups of polyphenols and the identification and quantification of individual components are presented. Here, a brief presentation of more recently published methods for grape and berry polyphenolic analyses is given with respect to their relationship to antioxidant activity and health benefits. 

Sharma U, Sharma K, Sharma N, Sharma S, Singh HP, Sinha AK, Microwave-assisted efficient extraction of different parts of hippophae rhamnoides for the comparative evaluation of antioxidant activity and quantification of its phenolic constituents by reverse-phase high-performance liquid chromatography (RPHPLC), J. Agric. Food Chem. 2008 56 374-379. 

Thus, antioxidant activity is a parameter that permits quantification of the capacity of a compound (natural or... 

Carotenoids are an important group of natural pigments with pronounced antioxidant activity, provitamin A factor, and many health benefits. The structure with conjugated double bonds governs mainly the proprieties of color, stability, detection, and quantification. With an increase of namral carotenoid markets, more different sources will be necessary and new technologies will be developed greener and cleaner, showing how promissory and profitable industrial sector can be. 

Tandem MS (DFS equipped with EI/F1/FD source) in conjunction with off-line direct inlet HPLC-UV was used for separation and quantification of isomeric antioxidants, C22H30O2S (MW 358 Scheme 6.3), as antioxidants in THF extracts of surgeons gloves [232]. Collision activation MS enabled differentiation between the three isomeric structures (Fig. 6.20). Quantification was achieved by chromatographic analysis of the isomeric species, which are not distinguishable by MS. On-line LC-MS facilitates this kind of analysis. 

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