title: Quantification of malondialdehyde by HPLC-FL - application to various biological samples.
abstract: Malondialdehyde (MDA) is stabile product of lipid peroxidation (LPO), and therefore MDA is frequently used as a biomarker of LPO. To determine MDA level in various biological samples (human plasma, fish liver tissue and cells in culture), we used an HPLC method with fluorescent detection based on 2-thiobarbituric acid (TBA) assay. The method was validated by the use of spiked pooled plasma samples. In tested concentration range (0.15-3.0 µmol/L) the method was linear (R(2)  = 0.9963), the between-day variability (coefficient of variations, CVs) was between 4.7 and 7.6%, the within-day variability CVs was between 2.6 and 6.4% and recovery was between 91.2 and 107.6%. The level of MDA in human plasma (healthy male, non-smokers, 46.3 ± 4.7 years; N = 38) was 2.2 ± 1.4 µmol/L; that in liver tissue of common carp (Cyprinus carpio; N = 12) was 0.02 ± 0.004 µmol/g tissue, and in cultured cells (human laryngeal carcinoma cells; N = 10) it was 0.18 ± 0.02 nmol/mg proteins. The HPLC-FL method is rapid, accurate and reliable to follow the extent of LPO in various biological samples, particularly in samples in which a low level of MDA is expected, such as cells in culture. Owing to the rapid analytical process and run time, it can be used for routine analysis of MDA in clinical laboratory.
authors.affiliation: University of Zagreb, Faculty of Pharmacy and Biochemistry, Department of Pharmaceutical Botany, Zagreb, Croatia.
authors.forename: Ana-Marija
authors.initials: AM
authors.lastname: Domijan
authors.forename: Jovica
authors.initials: J
authors.lastname: Ralić
authors.forename: Sandra
authors.initials: S
authors.lastname: Radić Brkanac
authors.forename: Lada
authors.initials: L
authors.lastname: Rumora
authors.forename: Tihana
authors.initials: T
authors.lastname: Žanić-Grubišić
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D016014:Linear Models; D008099:Liver / Q000737:chemistry; D008315:Malondialdehyde / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013858:Thiobarbiturates / Q000737:chemistry / Q000378:metabolism
chemical_list: D013858:Thiobarbiturates; D008315:Malondialdehyde; C029684:thiobarbituric acid
keywords: cells in culture; lipid peroxidation; liver homogenate; plasma; thiobarbituric acid assay