Lipids

, Volume 26, Issue 10, pp 853–856

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

  • Zhen-Yue Jiang
  • Alison C. S. Woollard
  • Simon P. Wolff
Method

DOI: 10.1007/BF02536169

Cite this article as:
Jiang, Z., Woollard, A.C.S. & Wolff, S.P. Lipids (1991) 26: 853. doi:10.1007/BF02536169

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.

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

Copyright information

© American Oil Chemists’ Society 1991

Authors and Affiliations

  • Zhen-Yue Jiang
    • 1
  • Alison C. S. Woollard
    • 1
  • Simon P. Wolff
    • 1
  1. 1.Toxicology Laboratory, Dept. of Clinical PharmacologyUniversity College and Middlesex School of MedicineLondonU.K.