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Measurement of malondialdehyde by high performance liquid chromatography with fluorescence detection

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Lipids

Abstract

A method has been developed to measure malondial-dehyde (MDA) in biological systems. MDA was reacted with 2-thiobarbituric acid (TBA) in the presence of butylated hydroxytoluene (BHT) to minimize formation of artifacts. Initial separation of the TBA-MDA adduct was accomplished by isobutanol extraction. Further elimination and separation of interfering substances was achieved by high performance liquid chromatography. The mobile phase consisted of a 1∶1 (v/v) mixture of methanol and water with 0.05% (w/v) tetrabutyl ammonium dihydrogen phosphate added as an ion pairing reagent. At a flow rate of 1 ml/min, the TBA-MDA adduct was eluted from a 15-cm, c-18, reversed phase column in approximately 4.9 min. The TBA-MDA adduct was quantitated with a fluorescence detector set at 515 nm excitation and 550 nm emission. Using this method, picomole quantities of MDA can be easily detected in plasma and liver samples.

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Abbreviations

BHT:

butylated hydroxytoluene

HCI:

hydrochloric acid

HPLC:

high performance liquid chromatography

MDA:

malondialdehyde

TBA:

2-thiobarbituric acid

TMP:

1,1,3,3-tetramethoxypropane

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Authors and Affiliations

  1. Department of Nutrition and Food Science and Graduate Center for Toxicology, University of Kentucky, 40506-0054, Lexington, KY

    Vickie L. Tatum, Charus Changchit & Ching K. Chow

Authors
  1. Vickie L. Tatum
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  2. Charus Changchit
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  3. Ching K. Chow
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Tatum, V.L., Changchit, C. & Chow, C.K. Measurement of malondialdehyde by high performance liquid chromatography with fluorescence detection. Lipids 25, 226–229 (1990). https://doi.org/10.1007/BF02535752

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  • DOI: https://doi.org/10.1007/BF02535752

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