Abstract
A reliable method for spectrophotometric determination of urinary malondialdehyde (MDA), according to the thiobarbituric acid (TBA) assay, is described. To account for matrix interference and differences in individual urine composition, standard addition procedure was applied. The method is adequately selective (LoQ = 0.09 μM in the presence of 0.1 M creatinine and 0.5 M urea) and reliable (within-day and between-day variability of less than 5 %). The mean level of urinary MDA was 1.52 ± 0.73 µM that is in good agreement with spectrofluorometric determination (1.20 ± 0.56 μM; p = 0.085) as well as with previous studies that used HPLC. Furthermore, it is demonstrated that MDA is stabile in urine at room temperature for 24 h and when stored at –20 °C for 6 months. The described method enables simple, rapid and cost-effective determination of urinary MDA as a relevant and non-invasive marker of “whole-body” oxidative stress. This work is licensed under a Creative Commons Attribution 4.0 International License .
Highlights
T HE dicarbonyl malondialdehyde (MDA) is one of the products of lipid peroxidation.[1,2] Lipid peroxidation is a process in biological systems initiated by an attack of reactive oxygen species (ROS) on polyunsaturated fatty acids, major constituents of plasma membranes
It is demonstrated that MDA is stabile in urine at room temperature for 24 h and when stored at –20 °C for 6 months
The results for urinary MDA obtained spectrophotometrically were verified spectrofluorometrically. These results indicate that the described spectrophotometric method can be used for determination of MDA in urine in order to follow the extent of “whole-body” oxidative stress
Summary
T HE dicarbonyl malondialdehyde (MDA) is one of the products of lipid peroxidation.[1,2] Lipid peroxidation is a process in biological systems initiated by an attack of reactive oxygen species (ROS) on polyunsaturated fatty acids, major constituents of plasma membranes. The first products of lipid peroxidation are unstable lipid peroxides that are further converted by consecutive reactions of oxidation, rearrangements and scission into more stable carbonyl compounds, among which is MDA.[2,3]. The condition of imbalance between the metabolic production of reactive species and the cellular ability to scavenge the reactive metabolites or repair the incurred damage is oxidative stress. This condition is connected to the development of various diseases, including cancer.[3,6] it is important to find reliable markers to follow the extent of oxidative stress. The level of MDA as a final product of lipid peroxidation in specific tissue is considered as a marker of oxidative stress in that particular tissue, and MDA level in plasma and urine represents “whole-body” oxidative stress.[4,7] Drury at al.[4] demonstrated by comparing plasma and urine samples that urinary MDA level is good marker of “whole-body” oxidative stress in populations without renal disease
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