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. 2022 Aug 29;12(9):811.
doi: 10.3390/metabo12090811.

Stability of Metabolomic Content during Sample Preparation: Blood and Brain Tissues

Maxim V Fomenko  1   2 , Lyudmila V Yanshole  1 , Yuri P Tsentalovich  1
Affiliations

Stability of Metabolomic Content during Sample Preparation: Blood and Brain Tissues

Maxim V Fomenko et al. Metabolites. .

Abstract

Thermal and enzymatic reactions can significantly change the tissue metabolomic content during the sample preparation. In this work, we evaluated the stability of metabolites in human whole blood, serum, and rat brain, as well as in metabolomic extracts from these tissues. We measured the concentrations of 63 metabolites in brain and 52 metabolites in blood. We have shown that metabolites in the extracts from biological tissues are stable within 24 h at 4 °C. Serum and whole blood metabolomes are also rather stable, changes in metabolomic content of the whole blood homogenate become apparent only after 1-2 h of incubation at 4 °C, and become strong after 24 h. The most significant changes correspond to energy metabolites: the concentrations of ATP and ADP decrease fivefold, and the concentrations of NAD, NADH, and NADPH decrease below the detectable level. A statistically significant increase was observed for AMP, IMP, hypoxanthine, and nicotinamide. The brain tissue is much more metabolically active than human blood, and significant metabolomic changes occur already within the first several minutes during the brain harvest and sample homogenization. At a longer timescale (hours), noticeable changes were observed for all classes of compounds, including amino acids, organic acids, alcohols, amines, sugars, nitrogenous bases, nucleotides, and nucleosides.

Keywords: NMR spectroscopy; human blood; metabolomics; rat brain; sample preparation.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Kinetics of metabolite concentrations in the whole blood homogenates during the incubation at 4 °C.
Figure 2
Figure 2
Kinetics of metabolite concentrations in the rat brain homogenates during the incubation at 4 °C.
Figure 3
Figure 3
General scheme of post-mortem metabolite transformations in rat brain homogenates.
See this image and copyright information in PMC

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