engleski

Quantification of malondialdehyde by HPLC-FL – application to various biological samples

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 (R2 = 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.

lipid peroxidation; thiobarbituric acid assay; plasma; cells in culture; liver homogenate

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