The ferric reducing ability of plasma (FRAP) as a measure of `'antioxidant power'': The FRAP assay

IFF Benzie, JJ Strain

Research output: Contribution to journalArticle

10475 Citations (Scopus)

Abstract

A simple, automated test measuring the ferric reducing ability of plasma, the FRAP assay, is presented as a novel method for assessing `'antioxidant power.'' Ferric to ferrous ion reduction at low pH causes a colored ferrous-tripyridyltriazine complex to form. FRAP values are obtained by comparing the absorbance change at 593 nm in test reaction mixtures with those containing ferrous ions in known concentration. Absorbance changes are linear over a wide concentration range with antioxidant mixtures, including plasma, and with solutions containing one antioxidant in purified form. There is no apparent interaction between antioxidants. Measured stoichiometric factors of Trolox, alpha-tocopherol, ascorbic acid, and uric acid are all 2.0; that of bilirubin is 4.0. Activity of albumin is very low. Within- and between-run CVs are <1.0 and <3.0%, respectively, at 100-1000 mu mol/liter. FRAP values of fresh plasma of healthy Chinese adults: 612-1634 mu mol/liter (mean, 1017; SD, 206; n = 141). The FRAP assay is inexpensive, reagents are simple to prepare, results are highly reproducible, and the procedure is straightforward and speedy. The FRAP assay offers a putative index of antioxidant, or reducing, potential of biological fluids within the technological reach of every laboratory and researcher interested in oxidative stress and its effects. (C) 1996 Academic Press, Inc.
LanguageEnglish
Pages70-76
JournalAnalytical Biochemistry
Volume239
Issue number1
Publication statusPublished - Jul 1996

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Assays
Antioxidants
Plasmas
Ions
Oxidative stress
alpha-Tocopherol
Uric Acid
Bilirubin
Ascorbic Acid
Albumins
Fluids

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abstract = "A simple, automated test measuring the ferric reducing ability of plasma, the FRAP assay, is presented as a novel method for assessing `'antioxidant power.'' Ferric to ferrous ion reduction at low pH causes a colored ferrous-tripyridyltriazine complex to form. FRAP values are obtained by comparing the absorbance change at 593 nm in test reaction mixtures with those containing ferrous ions in known concentration. Absorbance changes are linear over a wide concentration range with antioxidant mixtures, including plasma, and with solutions containing one antioxidant in purified form. There is no apparent interaction between antioxidants. Measured stoichiometric factors of Trolox, alpha-tocopherol, ascorbic acid, and uric acid are all 2.0; that of bilirubin is 4.0. Activity of albumin is very low. Within- and between-run CVs are <1.0 and <3.0{\%}, respectively, at 100-1000 mu mol/liter. FRAP values of fresh plasma of healthy Chinese adults: 612-1634 mu mol/liter (mean, 1017; SD, 206; n = 141). The FRAP assay is inexpensive, reagents are simple to prepare, results are highly reproducible, and the procedure is straightforward and speedy. The FRAP assay offers a putative index of antioxidant, or reducing, potential of biological fluids within the technological reach of every laboratory and researcher interested in oxidative stress and its effects. (C) 1996 Academic Press, Inc.",
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The ferric reducing ability of plasma (FRAP) as a measure of `'antioxidant power'': The FRAP assay. / Benzie, IFF; Strain, JJ.

In: Analytical Biochemistry, Vol. 239, No. 1, 07.1996, p. 70-76.

Research output: Contribution to journalArticle

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