The assays used for assessing antioxidant capacities of herbal products and foods

Öz

In this study, the assays used for antioxidant capacity of herbal products and foods were described comparatively each other. Various assays have been used to evaluation of antioxidant capacity of herbal products and foods. These assays should be classified into two types: assays based on hydrogen atom transfer (HAT) reactions and assays based on electron transfer (ET). HAT based assays include oxygen radical absorbance capacity (ORAC), total radical trapping antioxidant parameter (TRAP), and crocin bleaching assays. ET-based assays include the total phenols assay by Folin-Ciocalteu reagent (FCR), Trolox equivalence antioxidant capacity (TEAC), ferric ion reducing antioxidant power (FRAP), “total antioxidant potential” assay using a Cu (II) complex as an oxidant, and DPPH assay. Electron transfer assays measure the reducing ability of the substrate (antioxidant) while hydrogen atom transfer assays measure the hydrogen donating ability of the substrate. It is clear that hydrogen atom donation is essential in the radical chain reaction stage of lipid peroxidation, therefore hydrogen transfer assays are relevant to the measurement of chain-breaking antioxidant capacity. In many cases, the antioxidant capacity or the ability to trap radicals, of a compound is related to the ease of hydrogen atom donation, not necessarily the redox potential of the compound. In general, assays that measure hydrogen atom transfer would be preferable to assays that measure electron transfer reactions. In the study, the use of multiple electron transfer assays is redundant. There are correlations between them, because these assays rely on the same redox reactions. When the assays measure reducing ability were used it is important to choose one that is widely reported and validated. Similarly, the ORAC assay between assays that measure hydrogen atom transfer has been extensively validated, reliable and provides useful information about the radical chain-breaking capacity of the sample.

Anahtar Kelimeler

• Antioxidant capacity, DPPH, FRAP, ORAC, TEAC

Bitkisel ürünlerin ve gıdaların antioksidan kapasitelerinin belirlenmesinde kullanılan yöntemler

Öz

Bu çalışmada bitkisel ürünler ve gıdaların antioksidan kapasitesini belirlemek için kullanılan yöntemler birbirleri ile karşılaştırmalı olarak tanımlanmıştır. Bitkisel ürünlerin ve gıdaların antioksidan kapasitesini değerlendirmek için çeşitli yöntemler kullanılmaktadır. Bu yöntemler elektron transfer (ET) ve hidrojen atom transfer (HAT) reaksiyonlarına dayanan yöntemler olmak üzere iki grup altında incelenebilir. HAT temelli yöntemler oksijen radikal absorbans kapasite (ORAC), toplam radikal yakalayıcı antioksidan parametre (TRAP) ve krosin beyazlatma yöntemlerini içermektedir. ET temelli yöntemler ise Folin-Ciocalteu ayıracı ile toplam Fenolik yöntemi (FCR), Troloks eşiti antioksidan kapasite (TEAC), demir iyonu indirgeyici antioksidan güç (FRAP), oksidan olarak bakır (II) kullanan toplam antioksidan potansiyel yöntemi (CUPRAC) ve DPPH yöntemini içermektedir. Elektron transfer yöntemleri substratın (antioksidan) indirgeyici yeteneğini ölçerken, hidrojen atom transfer yöntemleri substratın hidrojen verebilme yeteneğini ölçer. Hidrojen atom vermenin lipit peroksidayonunun radikal zincir reaksiyon evresinde önemli olduğu açıktır. Bu nedenle hidrojen transfer yöntemleri zincir-kırma antioksidan kapasitesinin ölçülmesi ile ilişkilidir. Birçok durumda bir bileşiğin antioksidan kapasitesi veya radikal yakalama yeteneği hidrojen atom verebilme kolaylığı ile alakalıdır, bileşiğin redoks potansiyeli gerekli değildir. Genellikle hidrojen atom transferi ölçen yöntemler elektron transfer reaksiyonlarını ölçen yöntemlere göre daha çok tercih edilmektedir. Çalışmada çok sayıda elektron transfer yönteminin kullanılması gereksizdir. Bu yöntemler benzer redoks reaksiyonlarına dayandığı için yöntemler arasında ilişki vardır. İndirgeyici kapasiteyi ölçen yöntemler kullanılacağı zaman yaygın olarak kullanılan ve geçerliliği yüksek olan yöntemi tercih etmek önemlidir. Benzer olarak, hidrojen atom transferi ölçen yöntemler arasında ORAC yöntemi büyük oranda geçerlilik kazanmıştır, güvenilirdir ve örneğin radikal zincir kırma kapasitesi hakkında kullanışlı bilgi vermektedir.

Anahtar Kelimeler

• Antioksidan kapasite, DPPH, FRAP, ORAC, TEAC

Kaynakça

1. Lichtenthaler, R., Marx, F. and Kind, O.M., Determination of antioxidative capacities using an enhanced total oxidant scavenging capacity (TOSC) assay, Eur. Food Res. Technol., 216,166–173, 2003.
2. Schoneich, C., Reactive oxygen species and biological aging: a mechanistic approach, Experimental Gerontology, 34(1), 19-34, 1999.
3. Aruoma, O.I., Free radicals, oxidative stress and antioxidants in human health and disease, J. Am. Oil Chem. Soc., 75(2), 199–212, 1998.
4. MacDonald-Wicks, L.K., Wood L.G and Garg, M.L., Methodology for the determination of biological antioxidant capacity in vitro: a review, J. Sci. Food Agric., 86, 2046–2056, 2006.
5. Price, J.A., Sanny, C.G., Shevlin, D., Application of manual assessment of oxygen radical absorbent capacity (ORAC) for use in high throughput assay of ‘‘total’’ antioxidant, activity of drugs and natural products, Journal of Pharmacological and Toxicological Methods, 54, 56 – 61, 2006.
6. Ames, B.N., Shigenaga, M.K., Hagen, T.M., “Oxidants, antioxidants, and the degenerative diseases of aging”, Proc. Natl. Acad. Sci. U.S.A., 90, 7915-7922, 1993.
7. Prior, R.L. and Cao, G., In vivo Total Antioxidant Capacity: Comparison of Different Analytical Methods, Free Radical Bio. Med., 27, 1173–1181, 1999.
8. Somogyi, A., Rosta, K., Pusztai, P., Tulassay Z. and Nagy, G., Antioxidant measurements, Physiol. Meas., 28, R41–R55, 2007.

9. Pellegrini, N., Serafini, M., Colombi, B., Del Rio, D., Salvatore, S., Bianchi, M. and Brighenti, F., Total antioxidant capacity of plant foods, beverages and oils consumed in Italy assessed by three different in vitro assays, J. Nutr., 133, 2812–2819, 2003.
10. Huang, D., Ou, B., Prior, R.L., The Chemistry behind Antioxidant Capacity Assays, J. Agric. Food Chem., 53, 1841-1856, 2005.
11. Decker, E.A., K. Wagner, Richards, M.P. and Shahidi, F., Measuring Antioxidant Effectiveness in Food, J. Agric. Food Chem., 53, 4303-4310, 2005.
12. Ellefson, W., Lilla Z. and Crowley, R., Quantification and Evaluation of Antioxidants in Food and Botanical Products, International Conference on Nutraceuticals and Functional Foods, November 5-8, Reno, Neveda, 2006.
13. Gillespie, K.M., Chae, J.M. and Ainsworth, E.A., Rapid measurement of total antioxidant capacity in plants, Nat. Protoc., 2 (4), 867-870, 2007.
14. Tomer, P.D., McLeman, L.D., Ohmine, S., Scherer, P.M., Murray, B.K., O’Neill, K.L., Comparison of the Total Oxyradical scavenging capacity and oxygen radical absorbance capacity antioxidant assays, J. Med. Food, 10 (2), 337- 344, 2007.
15. Cao, G. and Prior, R.L., Comparison of different analytical methods for assessing total antioxidant capacity of human serum, Clin. Chem., 44, 1309–1315, 1998.
16. Pérez-Jiménez, J. and Saura-Calixto, F., Effect of solvent and certain food constituents on different antioxidant capacity assays, Food Res. Int., 39, 791–800, 2006.
17. Ou, B., Hampsch-Woodill, M., Prior, R.L., Development and validation of an improved oxygen radical absorbance capacity using fluorescein as the fluorescent probe, J. Agric. Food Chem., 49, 4619–4626, 2001.
18. Wu, X., Beecher, G.R., Holden, J.M., Haytowitz, D.B., Gebhardt, S.E., Prior, R.L., Lipophilic and hydrophilic antioxidant capacities of common foods in the United States, J. Agric. Food Chem., 52, 4026-4037, 2004.
19. Bonanni, A., Campanella, L., Gatta, T., Gregori, E., Tomassetti M., Evaluation of the antioxidant and prooxidant properties of several commercial dry spices by different analytical methods, Food Chem., 102, 751–758, 2007.
20. Ou, B., Hampsch-Woodill, M., Flanagan, J., Deemer, E.K., Prior, R.L., Huang, D., Novel fluorometric assay for hydroxyl radical prevention capacity using fluorescein as the probe, J. Agric. Food Chem., 50, 2772-2777, 2002.
21. Thaipong, K., Boonprakob, U., Crosby, K., Cisneros-Zevallos, L., Byrne, D.H., Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimating antioxidant activity from guava fruit extracts, J. Food Compos. Anal., 19, 669–675, 2006.
22. Ghiselli, A., Serafini, M., Natella, F., Scaccini, C., Total antioxidant capacity as a tool to assess redox status: critical view and experimental data. Free Radical Biol. Med., 29, 1106- 1114, 2000.
23. Prior, R.L., Wu, X., Schaich, K., Standardized Methods for the Determination of Antioxidant Capacity and Phenolics in Foods and Dietary Supplements, J. Agric. Food Chem., 53, 4290- 4302, 2005.
24. Lussignoli, S., Fraccaroli, M., Andrioli, G., Brocco, G. and Bellavite, P., A microplate-based colorimetric assay of the total peroxyl radical trapping capability of human plasma, Anal. Biochem., 269, 38–44. 1999.
25. Magalhães, L.M., Segundo, M.A., Reis, S., Lima, J.L.F.C., Methodological aspects about in vitro evaluation of antioxidant properties, Anal. Chim. Acta, 613, 1–19, 2008.
26. Singleton, V.L., Orthofer, R., Lamuela-Raventos, R.M., Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent, Methods Enzymol., 299, 152-178, 1999.
27. Miller, N.J., Rice-Evans, C., Davies, M.J., Gopinathan, V., Milner, A., A novel method for measuring antioxidant capacity and its application to monitoring the antioxidant status in premature neonates, Clin. Sci., 84, 407–412, 1993.
28. Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., Rice-Evans, C., Antioxidant activity applying an improved ABTS radical cation decolorization assay, Free Radic. Biol. Med., 26, 1231–1237, 1999.
29. Benzie, İ.F.F. and Strain, J.J., The Ferric Reducing Ability of Plasma (FRAP) as a Measure of “Antioxidant Power”: The FRAP Assay , Anal. Biochem., 239 (1), 70-76, 1996.
30. Molyneux, P., The use of the stable free radical diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity, Songklanakarin J. Sci. Technol., 26 (2), 211- 219, 2004.
31. Frankel, E.N. and Meyer, A.S., The problems of using one-dimensional methods to evaluate multifunctional food and biological antioxidants, J. Sci. Food Agr., 80, 1925-1941, 2000.
32. Scalzo, R.L., Organic acids influence on DPPH scavenging by ascorbic acid, Food Chem., 107, 40–43, 2008.
33. Aruoma, O.I., Methodological considerations for characterizing potential antioxidant actions of bioactive components in plant foods, Mutat. Res. Fundam. Mol., 523, 9-20, 2003.