Validation of methyl malondialdehyde as internal standard for malondialdehyde detection by capillary electrophoresis

Rita Paroni, Isabella Fermo, Giuliana Cighetti

Research output: Contribution to journalArticle

15 Citations (Scopus)

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 20cm × 50 μm uncoated capillaries at 20°C, using 25 mmol/L borax (pH 9.3) and 5 mmol/L tetradecyltrimethylammonium bromide as running buffer. The applied voltage was -4 kV (about 8 μA), 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 1 h at 60°C in the presence of 1 mol/L NaOH. The detection threshold was 0.2 μmol/L in microsomes and 0.4 μmol/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.

Original languageEnglish
Pages (from-to)92-98
Number of pages7
JournalAnalytical Biochemistry
Volume307
Issue number1
DOIs
Publication statusPublished - Aug 1 2002

Fingerprint

Capillary electrophoresis
Capillary Electrophoresis
Malondialdehyde
Plasma (human)
Liver Microsomes
Microsomes
Gas chromatography
Liver
Gas Chromatography-Mass Spectrometry
Dilution
Free Radicals
Mass spectrometry
Rats
Reactive Oxygen Species
Buffers
Proteins
Oxygen
Detectors
Plasmas
Electric potential

Keywords

  • Capillary electrophoresis
  • Malondialdehyde
  • Methyl malondialdehyde
  • Microsomes
  • Plasma

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Validation of methyl malondialdehyde as internal standard for malondialdehyde detection by capillary electrophoresis. / Paroni, Rita; Fermo, Isabella; Cighetti, Giuliana.

In: Analytical Biochemistry, Vol. 307, No. 1, 01.08.2002, p. 92-98.

Research output: Contribution to journalArticle

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