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

J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Sep 1:1029-1030:205-212. doi: 10.1016/j.jchromb.2016.07.013. Epub 2016 Jul 7.


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 30fmol (on-column). Intra- and inter-run coefficients of variation (CVs) were <2% and ∼10% respectively. The derivative was stable for >36h 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<0.02). The LC-MS method provides high sensitivity and high reproducibility for quantifying MDA in human plasma. The simple sample preparation and rapid analysis time (5x faster than LC-UV) offers high throughput for large-scale clinical applications.

Keywords: 3-Nitrophenylhydrazine; Derivatization; Isotope-labeling; LC/MRM–MS; Malondialdehyde; Plasma.

Publication types

  • Evaluation Study

MeSH terms

  • Chromatography, High Pressure Liquid / methods*
  • Depressive Disorder, Major / blood
  • Female
  • Humans
  • Limit of Detection
  • Male
  • Malondialdehyde / blood*
  • Phenylhydrazines / chemistry
  • Pilot Projects
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Tandem Mass Spectrometry / methods


  • 3-nitrophenylhydrazine
  • Phenylhydrazines
  • Malondialdehyde