Validation of methyl malondialdehyde as internal standard for malondialdehyde detection by capillary electrophoresis
Articolo
Data di Pubblicazione:
2002
Citazione:
Validation of methyl malondialdehyde as internal standard for malondialdehyde detection by capillary electrophoresis / R. Paroni, I. Fermo, G.M. Cighetti. - In: ANALYTICAL BIOCHEMISTRY. - ISSN 0003-2697. - 307:1(2002 Aug 01), pp. 92-98.
Abstract:
The aim of this study was to validate, by capillary electrophoresis, the use of synthesized methyl malondialdehyde as the internal standard for the direct quantification of free and total (free+bound) malondialdehyde in biological samples. All analyses were performed in 20 cm x 50 microm uncoated capillaries at 20 degrees C, using 25 mmol/L borax (pH 9.3) and 5 mmol/L tetradecyltrimethylammonium bromide as running buffer. The applied voltage was -4kV (about 8 microA), the detector being set at 260 nm for a total run time of 8 min per sample. Free malondialdehyde was evaluated after acetonitrile extraction, while the samples evaluated for total malondialdehyde were, before extraction, hydrolyzed for 1h at 60 degrees C in the presence of 1 mol/L NaOH. The detection threshold was 0.2 micromol/L in microsomes and 0.4 micromol/L in plasma. As an application of the method, three pools of rat liver microsomes were quantified before (0.35+/-0.1 and 1.1+/-0.5 nmol/mg protein, free and total malondialdehyde, respectively, mean+/-SD) and after lipoperoxidation induction using systems able to generate oxygen free radicals (18.4+/-3.2 and 19.7+/-2.0 nmol/mg protein). The results were confirmed by isotopic dilution gas chromatography-mass spectrometry, used as the reference method. The feasibility of capillary electrophoresis for malondialdehyde determination in normal and pathological human plasma was also investigated.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
Capillary electrophoresis ; Malondialdehyde ; Methyl malondialdehyde ; Microsomes ; Plasma
Elenco autori:
R. Paroni, I. Fermo, G.M. Cighetti
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