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.

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.

Free and Total Malondialdehyde Assessment in Biological Matrices by Gas Chromatography±Mass Spectrometry: What Is Needed for an Accurate Detection

G. Cighetti
Primo
;
R.Paroni
Penultimo
;
P. Allevi
Ultimo
1999

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.
free and total malondialdehyde; human plasma; GC–MS.
Settore BIO/10 - Biochimica
Settore BIO/12 - Biochimica Clinica e Biologia Molecolare Clinica
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/181309
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