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Lipid hydroperoxide measurement by oxidation of Fe 2+ in the presence of xylenol orange. Comparison with the TBA assay and an iodometric method
Author(s) -
Jiang ZhenYue,
Woollard Alison C. S.,
Wolff Simon P.
Publication year - 1991
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02536169
Subject(s) - xylenol orange , cumene hydroperoxide , chemistry , lipid peroxidation , iodometry , thiobarbituric acid , chromatography , liposome , tbars , nuclear chemistry , antioxidant , photochemistry , organic chemistry , biochemistry , catalysis
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 Fe 2+ under acidic conditions. Fe 3+ forms a chromophore with xylenol orange which absorbs strongly at 560 nm, yielding an apparent E 560 (for H 2 O 2 , butyl hydroperoxide and cumene hydroperoxide) of 4.3×10 4 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 hydroperoxide per se .