Amperometric biosensor based on a functionalized gold electrode for the detection of antioxidants.

A method for the electrochemical detection of antioxidants has been developed, which is based on a radical measurement with a cytochrome c modified electrode. A controlled enzymatic production system for superoxide radicals based on xanthine oxidase was used. The addition of antioxidants facilitated the decomposition of the radical in addition to the spontaneous dismutation. The steady-state of superoxide generation and decomposition was thus shifted to a new situation due to the higher decomposition rate after antioxidant addition. This resulted in a decreased current level at the electrode. Antioxidant activity could be quantified from the response of the sensor electrode by the percentage of the signal decrease. The 50% inhibition value (IC(50)) for different antioxidants was calculated and the antioxidant activity of numerous substances was compared. Thus, a hierarchy of superoxide radical scavenging abilities of flavonoids was established: flavanols>flavonols>flavones>flavonones>isoflavonones.

[1]  J. Rowley,et al.  Dietary bioflavonoids induce cleavage in the MLL gene and may contribute to infant leukemia. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Sawai,et al.  NMR Analytical Approach To Clarify the Antioxidative Molecular Mechanism of Catechins Using 1,1-Diphenyl-2-picrylhydrazyl. , 1998, Journal of agricultural and food chemistry.

[3]  F. Saura-calixto,et al.  Antioxidant activity of dietary polyphenols as determined by a modified ferric reducing/antioxidant power assay. , 2000, Journal of agricultural and food chemistry.

[4]  P. Calsou,et al.  Repair of oxidative DNA damage in vitro: a tool for screening antioxidative compounds. , 1999, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[5]  N. Miller,et al.  Are whole extracts and purified glucosinolates from cruciferous vegetables antioxidants? , 1996, Free radical research.

[6]  C. Fraga,et al.  Flavonoids as antioxidants evaluated by in vitro and in situ liver chemiluminescence. , 1987, Biochemical pharmacology.

[7]  J J Strain,et al.  The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay. , 1996, Analytical biochemistry.

[8]  S. Ostrovidov,et al.  Screening of new antioxidant molecules using flow cytometry. , 2000, Journal of Medicinal Chemistry.

[9]  Á. Ríos,et al.  A poly(vinyl choloride) graphite composite electrode for flow-injection amperometric determination of antioxidants , 1999 .

[10]  B. Halliwell,et al.  Antioxidants in human health and disease. , 1996, Annual review of nutrition.

[11]  R. Prior,et al.  In vivo total antioxidant capacity: comparison of different analytical methods. , 1999, Free radical biology & medicine.

[12]  C. Berset,et al.  Use of a Free Radical Method to Evaluate Antioxidant Activity , 1995 .

[13]  R. Liu,et al.  Nutrition: Antioxidant activity of fresh apples , 2000, Nature.

[14]  A. Vercesi,et al.  Mitochondrial damage induced by conditions of oxidative stress. , 1999, Free radical biology & medicine.

[15]  H. Alessio,et al.  Oxygen-radical absorbance capacity assay for antioxidants. , 1993, Free radical biology & medicine.

[16]  L. Marquardt,et al.  Green tea extract and its polyphenols markedly inhibit luminol-dependent chemiluminescence activated by peroxynitrite or SIN-1. , 2000, Luminescence : the journal of biological and chemical luminescence.

[17]  Y. Katsumura,et al.  Interaction of hydrated electron with dietary flavonoids and phenolic acids: rate constants and transient spectra studied by pulse radiolysis. , 1999, Free radical biology & medicine.

[18]  L. Magnani,et al.  Spectrophotometric measurement of antioxidant properties of flavones and flavonols against superoxide anion , 2000 .

[19]  E. Cadenas,et al.  Biochemistry of oxygen toxicity. , 1989, Annual review of biochemistry.

[20]  L. Packer,et al.  alpha-Lipoic acid as a biological antioxidant. , 1995, Free radical biology & medicine.

[21]  N. Sugihara,et al.  Anti- and pro-oxidative effects of flavonoids on metal-induced lipid hydroperoxide-dependent lipid peroxidation in cultured hepatocytes loaded with alpha-linolenic acid. , 1999, Free radical biology & medicine.

[22]  R. Zhang,et al.  Dietary flavonoids, quercetin, luteolin and genistein, reduce oxidative DNA damage and lipid peroxidation and quench free radicals. , 1997, Cancer letters.

[23]  Y. Naguib.  A fluorometric method for measurement of oxygen radical-scavenging activity of water-soluble antioxidants. , 2000, Analytical biochemistry.

[24]  W. Pryor,et al.  A rapid screening test to determine the antioxidant potencies of natural and synthetic antioxidants , 1993 .

[25]  R. Prior,et al.  Antioxidant and prooxidant behavior of flavonoids: structure-activity relationships. , 1997, Free radical biology & medicine.

[26]  F. Scheller,et al.  Superoxide Dismutase Activity Measurement Using Cytochrome c-Modified Electrode. , 1999, Analytical Chemistry.

[27]  C. Rice-Evans,et al.  A novel method for measuring antioxidant capacity and its application to monitoring the antioxidant status in premature neonates. , 1993, Clinical science.

[28]  L. Skibsted,et al.  Regeneration of phenolic antioxidants from phenoxyl radicals: an ESR and electrochemical study of antioxidant hierarchy. , 1999, Free radical research.

[29]  Fred Lisdat,et al.  Superoxide sensor based on cytochrome c immobilized on mixed-thiol SAM with a new calibration method , 2002 .

[30]  I. Popov,et al.  Photochemiluminescent detection of antiradical activity: II. Testing of nonenzymic water-soluble antioxidants. , 1994, Free radical biology & medicine.

[31]  E. Lien,et al.  Carcinogenicity, mutagenicity and cancer preventing activities of flavonoids: a structure-system-activity relationship (SSAR) analysis. , 1994, Progress in drug research. Fortschritte der Arzneimittelforschung. Progres des recherches pharmaceutiques.

[32]  K. Krause.  Professional phagocytes: predators and prey of microorganisms. , 2000, Schweizerische medizinische Wochenschrift.

[33]  V. Palace,et al.  Antioxidant potentials of vitamin A and carotenoids and their relevance to heart disease. , 1999, Free radical biology & medicine.

[34]  F. Regoli,et al.  A rapid gas chromatographic assay for determining oxyradical scavenging capacity of antioxidants and biological fluids. , 1998, Free radical biology & medicine.

[35]  N. Chandel,et al.  Mitochondrial reactive oxygen species trigger hypoxia-induced transcription. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[36]  B. Ge,et al.  An electrochemical method for quantification of the radical scavenging activity of SOD , 1999 .

[37]  G. Mazza,et al.  Assessing antioxidant and prooxidant activities of phenolic compounds. , 2000, Journal of agricultural and food chemistry.

[38]  P. Yáñez‐Sedeño,et al.  Analytical Applications of Cylindrical Carbon Fiber Microelectrodes. Simultaneous Voltammetric Determination of Phenolic Antioxidants in Food , 1995 .

[39]  I. Fridovich.  Superoxide Anion Radical (O·̄2), Superoxide Dismutases, and Related Matters* , 1997, The Journal of Biological Chemistry.

[40]  P. Abuja,et al.  Prooxidant and antioxidant properties of Trolox C, analogue of vitamin E, in oxidation of low-density lipoprotein. , 1999, Free radical research.

[41]  M. Gordon.  Dietary antioxidants in disease prevention. , 1996, Natural product reports.

[42]  H. Bui,et al.  Quantitative structure-activity relationship analysis of phenolic antioxidants. , 1999, Free radical biology & medicine.

[43]  S. Chevion,et al.  The use of cyclic voltammetry for the evaluation of antioxidant capacity. , 2000, Free radical biology & medicine.

[44]  B. Marjanović,et al.  Flavonoids as Antioxidants , 1994 .

[45]  P. Hollman,et al.  Absorption, metabolism and health effects of dietary flavonoids in man. , 1997, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.