Measurement of total and free malondialdehyde by gas–chromatography mass spectrometry – comparison with high-performance liquid chromatography methology. (August 2013)

• Record Type: Journal Article
• Authors: Zelzer, S.; Oberreither, R.; Bernecker, C.; Stelzer, I.; Truschnig-Wilders, M.; Fauler, G.
• Is Part Of: Free radical research. Volume 47:Number 8(2013:Aug.)
• Page Start: 651
• Page End: 656
• Publication Date: 2013-08
• Subjects: Free radicals (Chemistry) -- Periodicals; Antioxidants -- Periodicals; Vitamin C -- Periodicals; Vitamin E -- Periodicals; 541.224
• DOI: 10.3109/10715762.2013.812205
• Languages: English
• ISSNs: 1071-5762

Abstract

<abstract> <title>Abstract</title> <p>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-d<sub>2</sub>) as internal standard. This stable and precise GC–MS method showed good linearity (r<sup>2</sup> = 0.999) and higher specificity and sensitivity than the HPLC method and was validated for both total MDA (<italic>t</italic>-MDA) and free MDA (<italic>f</italic>-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 <italic>t</italic>-MDA and 0.039–0.625 μmol/L for <italic>f</italic>-MDA). Although comparison of <italic>t</italic>-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.</p> </abstract>