Free and Total Malondialdehyde Assessment in Biological Matrices by Gas Chromatography–Mass Spectrometry: What Is Needed for an Accurate Detection
✍ Scribed by Giuliana Cighetti; Sandra Debiasi; Rita Paroni; Pietro Allevi
- Publisher
- Elsevier Science
- Year
- 1999
- Tongue
- English
- Weight
- 89 KB
- Volume
- 266
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
A method to determine free and total (free and bound) malondialdehyde (MDA) in fresh human plasma, or in rat liver microsomes, using selected ion monitoring (SIM) gas chromatography-mass spectrometry in the electron impact mode was set up. The dideuterated internal standard, 3-hydroxy[1,3-2 H 2 ]-2-propenal (dMDA), was added to the biological samples before their analytical manipulation. To detect free MDA the samples were reacted under mild conditions (25°C, pH 4.0, 30 min) with phenylhydrazine (PH), affording 1-phenyl-1H-pyrazole and its 3,5-dideuterated isotopomer. For the evaluation of total MDA level the plasma or microsomes were subjected, before the derivatization step, to hydrolysis in the presence of 1 M NaOH under preestablished conditions. This method offers several advantages such specificity, precision (within-day CV 2.0%, between-day CV 2.1%), linearity (0.01-15 M) and high sensitivity (5 pmol injected). The recovery of known added MDA amounts from plasma and microsomes, hydrolyzed or not, accounted for 98 ؎ 0.6%. The free MDA levels found in the plasma and microsomes were 0.14 ؎ 0.03 M and 0.048 ؎ 0.006 nmol/mg protein, respectively. The total MDA levels were 1.3 ؎ 0.07 M in plasma and 0.36 ؎ 0.04 nmol/mg protein in the microsomes.