Biomedical Chemistry: Research and Methods 2024, 7(2), e00215

Influence of Derivatization Temperature and Butylated Hydroxytoluene on the Content of Malondialdehyde in Liver Homogenates

A.D. Konev1*, I.V. Aksenov1, V.A. Tutelyan1,2

1Federal Research Centre of Nutrition, Biotechnology and Food Safety,
2/14 Ustinsky pas., Moscow, 109240 Russia; *e-mail: konevtonyion@gmail.com
2I.M. Sechenov First Moscow State Medical University, 8 Trubetskaya str., Moscow, 119048 Russia

Keywords: lipid peroxidation; malondialdehyde; derivatization temperature; butylated hydroxytoluene; 2-thiobarbituric acid; liver homogenate.

DOI:10.18097/BMCRM00215

The whole version of this paper is available in Russian.

Malondialdehyde (MDA) is a product of lipid peroxidation that is widely used as a marker of oxidative stress in biomedical research. Detectable levels of MDA can vary significantly, which may be due to its formation in vitro during sample preparation. The purpose of the work was to analyze the methodological reasons for overestimating the malondialdehyde content in the liver and to find approaches to eliminate the flaws of the method. The amount of MDA was estimated by its derivatization with 2-thiobarbituric acid (TBA) with subsequent analysis by HPLC with fluorimetric detection. Increasing the derivatization temperature had no significant effect on the intensity of MDA-TBA complex formation when standard MDA solutions were used, but led to a sharp increase in its content in liver homogenates, which was dose-dependently prevented by the inclusion of butylhydroxytoluene (BHT) in the reaction mixture. The results obtained may be in demand for the development of methods for the analysis of MDA in organs and tissues, as well as for the interpretation of relevant data from biomedical studies.
Figure 1. Effect of reaction mixture incubation temperature on MDA content using standard MDA solution and liver homogenate (M±m; n=3). * - p<0.05 significant differences compared to the result obtained at the previous step of temperature increase.
Figure 2. Effect of BHT on MDA content in liver homogenate at 90°C incubation temperature of the reaction mixture 90°С (M±m; n=6). * - p<0.05 significant differences compared to the result obtained at the previous step of increasing BHT concentration.

FUNDING

This work was carried out using subsidies for the implementation of a state task in the Russian Federation (research No. FGMF-2023-0006).

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