A specific, accurate, and sensitive measure of total plasma malondialdehyde by HPLC[S]

Malondialdehyde (MDA) is one of the most commonly reported biomarkers of lipid peroxidation in clinical studies. The reaction of thiobarbituric acid (TBA) with MDA to yield a pink chromogen attributable to an MDA-TBA2 adduct is a common assay approach with products being quantified by ultraviolet-Vi...

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Bibliographic Details
Published in:Journal of lipid research 2013-03, Vol.54 (3), p.852-858
Main Authors: Moselhy, Hamdy F., Reid, Raymond G., Yousef, Saeed, Boyle, Susanne P.
Format: Article
Language:English
Subjects:
assay bias
assay specificity
Chromatography, High Pressure Liquid - methods
high-performance liquid chromatography
Humans
Lipid Peroxidation
Malondialdehyde - blood
Mass Spectrometry
Methods
Thiobarbituric Acid Reactive Substances - metabolism
thiobarbituric acid-reactive substance
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Summary:Malondialdehyde (MDA) is one of the most commonly reported biomarkers of lipid peroxidation in clinical studies. The reaction of thiobarbituric acid (TBA) with MDA to yield a pink chromogen attributable to an MDA-TBA2 adduct is a common assay approach with products being quantified by ultraviolet-Vis assay as nonspecific TBA-reactive substances (TBARS) or chromatographically as MDA. The specificity of the TBARS assay was compared with both chromatographic assays for total plasma MDA. The levels of total plasma MDA were significantly lower than the plasma TBARS in each of the samples examined, and interestingly, the interindividual variation apparent in the level of plasma MDA was not evident in the plasma TBARS assay. Each of the four online chromatographic detectors yielded a precise, sensitive, and accurate determination of total plasma MDA, and selected-ion monitoring was the most-accurate assay (101.3%, n = 4). The online diode array detectors provided good assay specificity (peak purity index of 999), sensitivity, precision, and accuracy. This research demonstrates the inaccuracy that is inherent in plasma TBARS assays, which claim to quantify MDA, and it is proposed that the TBARS approach may limit the likelihood of detecting true differences in the level of lipid peroxidation in clinical studies.
ISSN:0022-2275
1539-7262
DOI:10.1194/jlr.D032698