Development and evaluation of a liquid chromatography–mass spectrometry method for rapid, accurate quantitation of malondialdehyde in human plasma

Abstract Malondialdhyde (MDA) is a commonly used marker of lipid peroxidation in oxidative stress. To provide a sensitive analytical method that is compatible with high throughput, we developed a multiple reaction monitoring-mass spectrometry (MRM-MS) approach using 3-nitrophenylhydrazine chemical derivatization, isotope-labeling, and liquid chromatography (LC) with electrospray ionization (ESI)–tandem mass spectrometry assay to accurately quantify MDA in human plasma. A stable isotope-labeled internal standard was used to compensate for ESI matrix effects. The assay is linear (R 2  = 0.9999) over a 20,000-fold concentration range with a lower limit of quantitation of 30 fmol (on-column). Intra- and inter-run coefficients of variation (CVs) were 36 h at 5 °C. Standards spiked into plasma had recoveries of 92–98%. When compared to a common LC-UV method, the LC–MS method found near-identical MDA concentrations. A pilot project to quantify MDA in patient plasma samples (n = 26) in a study of major depressive disorder with winter-type seasonal pattern (MDD-s) confirmed known associations between MDA concentrations and obesity (p