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Lipid hydroperoxide measurement by oxidation of Fe2+ in the presence of xylenol orange. Comparison with the TBA assay and an iodometric method

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Lipids

Abstract

Study of the role of hydroperoxides and lipid peroxidation in disease requires simple and sensitive methods for direct hydroperoxide measurement. We report on a technique for measuring hydroperoxide which relies upon the rapid hydroperoxide-mediated oxidation of Fe2+ under acidic conditions. Fe3+ forms a chromophore with xylenol orange which absorbs strongly at 560 nm, yielding an apparent E560 (for H2O2, butyl hydroperoxide and cumene hydroperoxide) of 4.3×104 M−1 cm−1. The assay was validated in a study of liposomal lipid peroxidation and shown to give results comparable with those obtained by an iodometric method or by measuring conjugated dienes. The assay involving thiobarbituric acid, by comparison, underestimates lipid peroxidation and does not measure hydroperoxideper se.

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Abbreviations

BHT:

butylated hydroxytoluene

CD:

conjugated diene

DTPA:

diethylenetriaminepentaacetic acid

FOX:

ferrous oxidation/xylenol orange

GSH:

reduced gluthathione

MDA:

malondialdehyde

PBS:

phosphate-buffered saline

TBA:

thiobarbituric acid

TBARM:

TBA-reactive material

TCA:

trichloroacetic acid

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Authors and Affiliations

  1. Toxicology Laboratory, Dept. of Clinical Pharmacology, University College and Middlesex School of Medicine, 5 University St., WC1E 6JJ, London, U.K.

    Zhen-Yue Jiang, Alison C. S. Woollard & Simon P. Wolff

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  1. Zhen-Yue Jiang
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  2. Alison C. S. Woollard
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  3. Simon P. Wolff
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Jiang, ZY., Woollard, A.C.S. & Wolff, S.P. Lipid hydroperoxide measurement by oxidation of Fe2+ in the presence of xylenol orange. Comparison with the TBA assay and an iodometric method. Lipids 26, 853–856 (1991). https://doi.org/10.1007/BF02536169

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  • DOI: https://doi.org/10.1007/BF02536169

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