Free and Total Malondialdehyde Assessment in Biological Matrices by Gas Chromatography±Mass Spectrometry: What Is Needed for an Accurate Detection
Articolo
Data di Pubblicazione:
1999
Citazione:
Free and Total Malondialdehyde Assessment in Biological Matrices by Gas Chromatography±Mass Spectrometry:
What Is Needed for an Accurate Detection / G. Cighetti, S. Debiasi, R.Paroni,P. Allevi. - In: ANALYTICAL BIOCHEMISTRY. - ISSN 0003-2697. - 266:2(1999 Jan 15), pp. 222-229.
Abstract:
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[1,3-2H2]-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 mM) 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 mM and 0.048 +/- 0.006 nmol/mg protein, respectively. The total MDA levels were 1.3 6 0.07 mM in plasma and 0.36 6 0.04 nmol/mg protein in the microsomes.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
free and total malondialdehyde; human
plasma; GC–MS.
Elenco autori:
G. Cighetti, S. Debiasi, R.Paroni,P. Allevi
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