TY - JOUR

T1 - Stability of Metabolomic Content during Sample Preparation: Blood and Brain Tissues

AU - Fomenko, Maxim V.

AU - Yanshole, Lyudmila V.

AU - Tsentalovich, Yuri P.

N1 - Funding Information: This research was funded by Russian Foundation for Basic Research, grant number 22-24-20035. Publisher Copyright: © 2022 by the authors.

PY - 2022/9

Y1 - 2022/9

N2 - 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.

AB - 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.

KW - human blood

KW - metabolomics

KW - NMR spectroscopy

KW - rat brain

KW - sample preparation

UR - http://www.scopus.com/inward/record.url?scp=85138645830&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/6ca4adb2-6405-3830-b062-f4c57eeee4a1/

U2 - 10.3390/metabo12090811

DO - 10.3390/metabo12090811

M3 - Article

C2 - 36144215

AN - SCOPUS:85138645830

VL - 12

JO - Metabolites

JF - Metabolites

SN - 2218-1989

IS - 9

M1 - 811

ER -