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Zelzer, S; Oberreither, R; Bernecker, C; Stelzer, I; Truschnig-Wilders, M; Fauler, G.
Measurement of total and free malondialdehyde by gas-chromatography mass spectrometry--comparison with high-performance liquid chromatography methology.
Free Radic Res. 2013; 47(8):651-656
Doi: 10.3109/10715762.2013.812205
Web of Science
PubMed
FullText
FullText_MUG
- Führende Autor*innen der Med Uni Graz
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Zelzer Sieglinde
- Co-Autor*innen der Med Uni Graz
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Bernecker Claudia
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Fauler Günter
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Stelzer Ingeborg
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Truschnig-Wilders Martini
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- Abstract:
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Malondialdehyde (MDA) is considered to be a biomarker for enzymatic degradation and lipid peroxidation of polyunsaturated fatty acids. Usually, MDA determination from different biological materials is performed by reaction with thiobarbituric acid (TBA) followed by high-performance liquid chromatography (HPLC) analysis and fluorometric detection. As this method lacks specificity and sensitivity, we developed a gas chromatography-mass spectrometry (GC-MS) method based on derivatization of MDA with 2,4-dinitrophenylhydrazine. Representative ions in negative ion chemical ionization (NICI) mode were recorded at m/z 204 for MDA and at m/z 206 for the deuterated analogon (MDA-d₂) as internal standard. This stable and precise GC-MS method showed good linearity (r² = 0.999) and higher specificity and sensitivity than the HPLC method and was validated for both total MDA (t-MDA) and free MDA (f-MDA). Within-day precisions were 1.8-5.4%, between-day precisions were 4.8-9.2%; and accuracies were between 99% and 101% for the whole calibration range (0.156-5.0 μmol/L for t-MDA and 0.039-0.625 μmol/L for f-MDA). Although comparison of t-MDA levels from GC-MS and HPLC results using Passing-Bablok regression analysis as well as Bland-Altman plot showed a correlation of the data, a tendency to increased results for the HPLC values was detectable, due to possible formation of unspecific products of the TBA reaction.
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Adult -
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Chromatography, High Pressure Liquid -
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Gas Chromatography-Mass Spectrometry -
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Humans -
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Malondialdehyde - blood
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GC-MS
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HPLC
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malondialdehyde
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oxidative stress
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total and free MDA