Abstract
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.
Highlights
Malondialdehyde (MDA) is one of the most commonly reported biomarkers of lipid peroxidation in clinical studies
Employing two chromatographic assays [HPLC-diode array detection (DAD)-fluoro and LC/MS-DAD], this research investigated the absolute levels of plasma MDA and compared these with the level of TBAreactive substances (TBARS) determined by UV spectrophotometry
It has been reported that acid precipitation of lipoprotein fractions reduces soluble TBARS components and enables a specific determination of lipid peroxidation without the need for HPLC [24]
Summary
Malondialdehyde (MDA) is one of the most commonly reported biomarkers of lipid peroxidation in clinical studies. Employing two chromatographic assays [HPLC-diode array detection (DAD)-fluoro and LC/MS-DAD], this research investigated the absolute levels of plasma MDA and compared these with the level of TBARS determined by UV spectrophotometry.
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