Modification of the deoxyribose test to detect strong iron binding

  • Izabela A. Sadowska-Bartosz University of Rzeszów, Department of Biochemistry and Cell Biology
  • Sabina Galiniak University of Rzeszow
  • Grzegorz Bartosz University of Łódź
Keywords: chelation, deoxyribose test, desferrioxamine, DETAPA, EDTA, Fenton reaction, hydrogen peroxide, hydroxyl radical, iron, superoxide


Deoxyribose test has been widely used for determination of reactivities of various compounds for the hydroxyl radical. The test is based on the formation of hydroxyl radical by Fe2+ complex in the Fenton reaction. We propose a modification of the deoxyribose test to detect strong iron binding, inhibiting participation of Fe2+ in the Fenton reaction, on the basis of examination of concentration dependence of deoxyribose damage on Fe2+concentration, at a constant concentration of a chelating agent.


Bhat VB, Sridhar GR, Madyastha KM (2001) Efficient scavenging of hydroxyl radicals and inhibition of lipid peroxidation by novel analogues of 1,3,7-trimethyluric acid. Life Sci 70: 381–393.

Chobot V (2010) Simultaneous detection of pro- and antioxidative effects in the variants of the deoxyribose degradation assay. J Agric Food Chem 58: 2088–2094. doi: 10.1021/jf902395k

De S, Adhikari S, Tilak-Jain J, Menon VP, Devasagayam TP (2008) Antioxidant activity of an aminothiazole compound: possible mechanisms. Chem Biol Interact 173: 215–223. doi: 10.1016/j.cbi.2008.03.011.

Dinis TC, Maderia VM, Almeida LM (1994) Action of phenolic derivatives (acetaminophen, salicylate, and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Arch Biochem Biophys 315: 161–169.

Genaro-Mattos TC, Dalvi LT, Oliveira RG, Ginani JS, Hermes-Lima M (2009) Reevaluation of the 2-deoxyribose assay for determination of free radical formation. Biochim Biophys Acta 1790: 1636–1642. doi: 10.1016/j.bbagen.2009.09.00

Guedes AC, Gião MS, Seabra R, Ferreira AC, Tamagnini P, Moradas-Ferreira P, Malcata FX (2013) Evaluation of the antioxidant activity of cell extracts from microalgae. Mar Drugs 11: 1256–1270. doi: 10.3390/md11041256.

Gutteridge JM, Halliwell B (1982) The role of the superoxide and hydroxyl radicals in the degradation of DNA and deoxyribose induced by a copper-phenanthroline complex. Biochem Pharmacol 31: 2801–2805.

Halliwell B, Gutteridge JM, Aruoma OI (1987) The deoxyribose method: a simple "test-tube" assay for determination of rate constants for reactions of hydroxyl radicals. Anal Biochem 165: 215–219.

Halliwell B, Gutteridge JM (1981) Formation of thiobarbituric-acid-reactive substance from deoxyribose in the presence of iron salts: the role of superoxide and hydroxyl radicals. FEBS Lett 128: 347–352.

Khokhar S, Apenten RKO (2003) Iron binding characteristics of phenolic compounds: some tentative structure–activity relations. Food Chem 81: 133–140.

Lapenna D, Ciofani G, Festi D, Neri M, Pierdomenico SD, Giamberardino MA, Cuccurullo F (2002) Antioxidant properties of ursodeoxycholic acid. Biochem Pharmacol 64: 1661–1667. doi:10.1016/S0006-2952(02)01391-6

Manoj VM, Aravindakumar CT (2003) Reaction of hydroxyl radicals with S-nitrosothiols: determination of rate constants and end product analysis. Org Biomol Chem 1: 1171–1175.

Mokdad-Bzeouich I, Kilani-Jaziri S, Mustapha N, Bedoui A, Ghedira K, Chekir-Ghedira L (2015) Evaluation of the antimutagenic, antigenotoxic, and antioxidant activities of Eriobotrya japonica leaves. Pharm Biol 53: 1786–1794. doi: 10.3109/13880209.2015.1008145

Qian SY, Buettner GR (1999) Iron and dioxygen chemistry is an important route to initiation of biological free radical oxidations: an electron paramagnetic resonance spin trapping study. Free Radic Biol Med 26: 1447–1456.

Rachmilovich-Calis S, Meyerstein N, Meyerstein D, Rachmilovich-Calis S, Meyerstein N, Meyerstein D (2009) A mechanistic study of the effects of antioxidants on the formation of malondialdehyde-like products in the reaction of hydroxyl radicals with deoxyribose. Chemistry 15: 7717–7723. doi: 10.1002/chem.200802272

Saran M, Michel C, Stettmaier K, Bors W (2000) Arguments against the significance of the Fenton reaction contributing to signal pathways under in vivo conditions. Free Radic Res 33: 567–579.

White JM, Flashka HA (1973) An automated procedure, with use of ferrozine, for assay of serum iron and total iron-binding capacity. Clin Chem 19: 526–528.

Winterbourn C (1991) Factors that influence the deoxyribose oxidation assay for Fenton reaction products. Free Radic Biol Med 11: 353–360.