Sensitive and selective quantification of free and total malondialdehyde in plasma using UHPLC-HRMS.
- Mendonça R Laboratory of Evolutionary Ecophysiology, Institute of Biology University of Neuchâtel, 2000 Neuchâtel, Switzerland.
- Gning O Laboratory of Evolutionary Ecophysiology, Institute of Biology University of Neuchâtel, 2000 Neuchâtel, Switzerland.
- Di Cesaré C Neuchâtel Platform of Analytical Chemistry, University of Neuchâtel, 2000 Neuchâtel, Switzerland.
- Lachat L Neuchâtel Platform of Analytical Chemistry, University of Neuchâtel, 2000 Neuchâtel, Switzerland.
- Bennett NC Department of Zoology and Entomology, University of Pretoria, Pretoria 0002, South Africa.
- Helfenstein F Laboratory of Evolutionary Ecophysiology, Institute of Biology University of Neuchâtel, 2000 Neuchâtel, Switzerland.
- Glauser G Neuchâtel Platform of Analytical Chemistry, University of Neuchâtel, 2000 Neuchâtel, Switzerland gaetan.glauser@unine.ch.
- 2017-07-12
Published in:
- Journal of lipid research. - 2017
2,4-dinitrophenylhydrazine
derivatization
fatty acid/oxidation
lipid peroxidation
mass spectrometry
oxidized lipids
quantitation
ultra-high-performance liquid chromatography-high-resolution mass spectrometry
Analytic Sample Preparation Methods
Animals
Blood Chemical Analysis
Chromatography, High Pressure Liquid
Humans
Limit of Detection
Malondialdehyde
Mass Spectrometry
Reference Standards
Reproducibility of Results
English
Quantification of malondialdehyde (MDA) as a marker of lipid peroxidation is relevant for many research fields. We describe a new sensitive and selective method to measure free and total plasmatic MDA using derivatization with 2,4-dinitrophenylhydrazine (DNPH) and ultra-HPLC-high-resolution MS. Free and total MDA were extracted from minute sample amounts (10 μl) using acidic precipitation and alkaline hydrolysis followed by acidic precipitation, respectively. Derivatization was completed within 10 min at room temperature, and the excess DNPH discarded by liquid-liquid extraction. Quantification was achieved by internal standardization using dideuterated MDA as internal standard. The method's lowest limit of quantification was 100 nM and linearity spanned greater than three orders of magnitude. Intra- and inter-day precisions for total MDA were 2.9% and 3.0%, respectively, and those for free MDA were 12.8% and 24.9%, respectively. Accuracy was 101% and 107% at low and high concentrations, respectively. In human plasma, free MDA levels were 120 nM (SD 36.26) and total MDA levels were 6.7 μM (SD 0.46). In addition, we show the applicability of this method to measure MDA plasma levels from a variety of animal species, making it invaluable to scientists in various fields.
- Language
-
- English
- Open access status
- bronze
- Identifiers
-
- DOI 10.1194/jlr.D076661
- PMID 28694297
- Persistent URL
- https://folia.unifr.ch/global/documents/291147
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