Abstract
Recent epidemiological studies have shown that there may be a link between oxidative stress and the development of several types of chronic diseases. Studies have also shown that diets rich in fruits and vegetables may decrease the incidence of cancer and other chronic diseases. The antioxidant activity of the phytochemicals these foods contain may be partially responsible for the decreased incidence of these diseases in people who regularly consume them. While there are several assays currently used to assess the antioxidant activity of phytochemicals and other antioxidant compounds, two are reviewed here in detail. The first is the oxygen radical absorbance capacity (ORAC) assay, which measures the decrease in fluorescence decay caused by antioxidants, and the second is the total oxyradical scavenging capacity (TOSC) assay, which measures the decrease in ethylene gas production caused by the inhibition of the thermal hydrolysis of ABAP (2,2′-Azobis(2-methyl-(propionamidine) dihydrochloride) by KMBA (α-keto-γ-(methylthio)butyric acid sodium salt) in the presence of antioxidant compounds. These two assays are discussed here, with an in depth review of their methodology and correlation.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Oxygen Radical Absorbance Capacity (ORAC) of Selected Foods (2007); Nutrient Data Laboratory, Agricultural Research Service, US Department of Agriculture, United States Department of Agriculture.
Ames BN, Gold LS, Willet WC (1995) The Causes and Prevention of cancer. Proc Natl Acad Sci USA 92:5258–5265
Ames BN, Shigenaga MK, Hagen TM (1993) Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci USA 90:7915–7922
Christen Y (2000) Oxidative stress and Alzheimer’s disease. Am J Clin Nutr 71:621S–629S
Diaz MN, Frei B, Keaney JF Jr (1997) Antioxidants and atherosclerotic heart disease. N Eng J Med 337:408–416
Lang AE, Lozano AM (1998) Parkinson’s disease. First of two parts. N Eng J Med 339:111–114
Packer L, Colman C (1999) The anti-oxidant miracle. Wiley, New York
Cao G, Prior RL (1998) Comparison of different analytical methods for assessing total antioxidant capacity of human serum. Clin Chem 44(6):1309–1315
Halliwell B, Gutteridge JMC, Cross CE (1992) Free Radicals, antioxidants, and human disease: where are we now? J Clin Lab Med 119:598–620
Tomer DP (2003) Measuring parameters that are important in cancer prevention and treatment: assessing antioxidant activity and angiogenesis. Brigham Young University, Master’s Thesis
McLeman LD (2004) Evidence of synergistic intracellular antioxidant networking and antioxidant regulation of cellular lipid peroxidation in HL-60 acute promyelogenous leukemia. Master’s Thesis. Brigham Young University. 1-37.
O’Neill K, Murray B (2002) Power plants. Woodland Publishing, Utah
Cao G, Booth SL, Sadowski JA, Prior RL (1998) Increases in human plasma antioxidant capacity after consumption of controlled diets high in fruit and vegetables. Am J Clin Nutr 68:1081–1087
Huang D, Ou B, Prior RL (2005) The chemistry behind antioxidant capacity assays. J Agric Food Chem 53:1841–1856
Ou B, Hampsch-Woodill M, Prior RL (2001) Development and validation of an improved oxygen radical absorbance capacity assay using fluorescein as the fluorescent probe. J Agric Food Chem 49(10):4619–4626
Cao G, Alessio H, Cutler R (1993) Oxygen-radical absorbance capacity assay for antioxidants. Free Radic Biol Med 14(3):303–311
Cao G, Sofic E, Prior RL (1996) Antioxidant capacity of tea and common vegetables. J Agric Food Chem 44:3426–3431
Huang D, Ou B, Hampsch-Woodill M, Flanagan J, Prior RL (2002) High-throughput assay of oxygen radical absorbance capacity (orac) using a multichannel liquid handling system coupled with a microplate fluorescence reader in 96-well format. J Agric Food Chem 50:4437–4444
Ou B, Huang D, Hampsch-Woodill M, Flanagan JA, Deemer EK (2002) Analysis of antioxidant of common vegetables employing oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays: a comparative study. J Agric Food Chem 50:3122–3128
Glazer AN (1990) Phycoerythrin fluorescence based assay for reactive oxygen species. Methods Enzymol 186:161–168
Cao G, Prior RL (1999) Measurement of oxygen radical absorbance capacity in biological samples. Methods Enzymol 299:50–62
Pellegrini N, Del Rio D, Colombi B, Bianchi M, Brighenti F (2003) Application of the 2, 2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation assay to a flow injection system for the evaluation of antioxidant activity of some pure compounds and beverages. J Agric Food Chem 51:260–264
Pulido R, Bravo L, Suara-Calixto F (2000) Antioxidant activity of dietary polyphenols as determined by a modified ferric reducing/antioxidant power assay. J Agric Food Chem 48:3396–3402
Aruoma OI (2003) Methodological considerations for characterizing potential antioxidant actions of bioactive components in plant foods. Mutat Res 523–524:9–20
Gorinstein S, Martin-Belloso O, Katrich E, Lojek A, Ciz M, Gligelmo-Miguel N, Haruenkit R, Park YS, Jung ST, Trakhtenberg S (2003) Comparison of the contents of the main biochemical compounds and the antioxidant activity of some Spanish olive oils as determined by four different radical scavenging tests. J Nutr Biochem 14:154–159
Yokozawa T, Cho EJ, Hara Y, Kitani K (2000) Antioxidative activity of green tea treated with radical initiator 2, 2’-azobis(2-amidinopropane) dihydrochloride. J Agric Food Chem. 48:5068–5073
Prior RL, Hoang H, Gu L, Wu X, Bacchiocca M, Howard L, Hampsch-Woodill M, Huang D, Ou B, Jacob R (2003) Assays for hydrophilic and lipophilic antioxidant capacity (oxygen radical absorbance capacity (ORAC(FL))) of plasma and other biological and food samples. J Agric Food Chem 51:3273–3279
Dugas AJ Jr, Castaneda-Acosta J, Bonin GC, Price KL, Fischer NH, Winston GW (2000) Evaluation of the total peroxyl radical-scavenging capacity of flavonoids: structure-activity relationships. J Nat Prod 63:327–331
Regoli F, Winston GW (1999) Quantification of total oxidant scavenging capacity of antioxidants for peroxynitrite, peroxyl radicals, and hydroxyl radicals. Toxicol Appl Pharmacol 156:96–105
Adapted from http://www.bmglabtech.com/application-notes/fluorescence-intensity/orac-148.cfm. Consulted 7/9/08.
Somogyi A, Rosta K, Pusztai P, Tulassay Z, Nagy G (2007) Antioxidant Measurements. Physiol Meas 28:R41–R55
Tomer DP, McLeman LD, Ohmine S, Scherer PM, Murray BK, O’Neill KL (2007) Comparison of the total oxyradical scavenging capacity and oxygen radical absorbance capacity assays. J Med Food 10(2):337–344
Lichtenthaler R, Marx F (2005) Total oxidant scavenging capacities of common european fruit and vegetable juices. J Agric Food Chem 53:103–110
Lichtenthaler R, Rodrigues RB, Maia JGS, Papagiannopoulos M, Fabricius H, Marx F (2005) Total oxidant scavenging capacities of Euterpe oleracea Mart. (Acai) fruits. Int J Food Sci Nutr 56(1):53–64
Company R, Serafim A, Cosson R, Camus L, Shillito B, Fiala-Medioni A, Bebianno MJ (2006) The effect of cadmium on antioxidant responses and the susceptibility to oxidative stress in the hydrothermal vent mussel Bathymodiolus azoricus. Marine Biol 148:817–825
Giessing AMB, Mayer LM (2004) Oxidative coupling during gut passage in marine deposit-feeding invertebrates. Limnol Oceanogr 49(3):716–726
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Garrett, A.R., Murray, B.K., Robison, R.A., O’Neill, K.L. (2010). Measuring Antioxidant Capacity Using the ORAC and TOSC Assays. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress II. Methods in Molecular Biology, vol 594. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-411-1_17
Download citation
DOI: https://doi.org/10.1007/978-1-60761-411-1_17
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-410-4
Online ISBN: 978-1-60761-411-1
eBook Packages: Springer Protocols