Communication Dans Un Congrès Année : 2024

Analytical strategies to study animal isotopic labelling experiment in metabolomic and lipidomic

Résumé

Animal isotopic labelling experiments are precious and scientifically necessary. These strategies improve our understanding of metabolism. Experimental protocols can be different: bolus of isotopic tracer, prolonged administration. Consequently, isotopic tracer incorporation differs in nature and quantity according to the metabolic pathways targeted. For high enrichment, mass spectrometry can be sufficient to detect labelled compounds. But, for low enrichment experiments, Isotop-Ratio Mass Spectrometry (IRMS) is completely suitable. This technic allows to measure the relative abundance of isotope in a sample with different configurations. First, samples can be analyzed directly in EA-IRMS (Elemental Analyser) which allows to determine the global enrichment. Secondly, enrichment can be measured for specific metabolites by gas chromatography (GC) coupled to IRMS detector. In this presentation, we focus on the incorporation of 13C lactate through 2 injections into mice. Livers and inguinal adipose tissues were studied. While labelled tissues or fluids are precious and to have a maximum of information on a single sample, we developed a dual extraction. This extraction permits to extract at the same time energetic metabolites and amino acids (in aqueous phase) and lipids (in organic phase). Neutral lipids, phospholipids and fatty acids of the lipids were separated by Thin Layer Chromatography (TLC) to be analyzed specifically. Different fractions sample (native tissue, organic and aqueous phase, and lipids TLC fraction) were analyzed by EA-IRMS. Amino acids and fatty acids were profiled by GC-C-IRMS and GC-MS. Enrichment have been observed on different fractions and metabolites. This preliminary work allows the comparison of different techniques to follow and quantified the incorporation of labelled metabolites in polar and apolar fraction extracted from different organs of a mouse.

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Dates et versions

hal-04651172 , version 1 (17-07-2024)

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  • HAL Id : hal-04651172 , version 1

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Océane Delos. Analytical strategies to study animal isotopic labelling experiment in metabolomic and lipidomic. 5th Annual Meeting of EpiLipidNET, EpilidNet, May 2024, Dresden (GERMANY), Germany. ⟨hal-04651172⟩
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