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A thermospray mass spectrometric assay for Fe-induced 4-hydroxynonenal in tissues

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Lipids

Abstract

A method is presented for the determination of 4-hydroxy-nonenal (HNE) in tissue homogenates followingin vitro lipid peroxidation induced by iron (Fe++). NHE is measured as the pentafluorobenzyl oxime derivative using liquid chromatography thermospray mass spectrometry.In vitro metabolism of HNEvia the glutathione/glutathione-S-transferase pathway was inhibited using iodoacetic and iodobenzoic acids. The assay has been used as an indicator of the peroxidizability of tissue samples from animals both adequate in and depleted of α-tocopherol. The concentrations of HNE produced in tissues taken from animals depleted of α-tocopherol were found to be up to 8 times higher than those taken from animals supplemented with α-tocopherol.

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Abbreviations

Fe++ :

iron(II) ion

GC/MS:

gas chromatography/mass spectrometry

GSH:

reduced glutathione

GSHT:

glutathione-S-transferase

HNE:

4-hydroxynonenal

HPLC:

high-performance liquid chromatography

IAA:

iodoacetic acid

IBA:

o-iodobenzoic acid

LC/MS:

liquid chromatography/thermospray mass spectrometry

PFB:

pentafluorobenzyl

PIPES:

piperazinediethanesulfonic acid

SIM:

selected ion monitoring

TBARS:

thiobarbituric acid reactive substances

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Author notes

  1. Dominic M. Walsh

    Present address: Division of Biochemistry, The Queen's University of Belfast, Northern Ireland

Authors and Affiliations

  1. Veterinary Sciences Division, Department of Agriculture For Northern Ireland, Stormont, BT4 3SD, Belfast, Northern Ireland, UK

    W. John Blanchflower, Dominic M. Walsh, Seamus Kennedy & D. Glenn Kennedy

Authors
  1. W. John Blanchflower
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  2. Dominic M. Walsh
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  3. Seamus Kennedy
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Blanchflower, W.J., Walsh, D.M., Kennedy, S. et al. A thermospray mass spectrometric assay for Fe-induced 4-hydroxynonenal in tissues. Lipids 28, 261–264 (1993). https://doi.org/10.1007/BF02536650

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  • DOI: https://doi.org/10.1007/BF02536650

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