Proton NMR enables the absolute quantification of aqueous metabolites and lipid classes in unique mouse liver samples

Aurélien Amiel (1, 2) , Marie Tremblay-Franco (3, 2) , Roselyne Gautier (3, 2) , Simon Ducheix (1) , Alexandra Montagner (1) , Arnaud Polizzi (4) , Laurent Debrauwer (3, 2) , Hervé Guillou (4) , Justine Bertrand-Michel (5, 6) , Cécile Canlet (3, 2)

1 ToxAlim - ToxAlim
2 National Infrastructure for Metabolomics and Fluxomics
3 E20 - Metatoul AXIOM
4 ToxAlim-TIM - Toxicologie Intégrative & Métabolisme
5 INSERM - Institut National de la Santé et de la Recherche Médicale
6 MetaToul Lipidomics

Hepatic metabolites provide valuable information on the physiological state of an organism, and thus, they are monitored in many clinical situations. Typically, monitoring requires several analyses for each class of targeted metabolite, which is time consuming. The present study aimed to evaluate a proton nuclear magnetic resonance (1H-NMR) method for obtaining quantitative measurements of aqueous and lipidic metabolites. We optimized the extraction protocol, the standard samples, and the organic solvents for the absolute quantification of lipid species. To validate the method, we analyzed metabolic profiles in livers of mice fed three different diets. We compared our results with values obtained with conventional methods and found strong correlations. The 1H-NMR protocol enabled the absolute quantification of 29 aqueous metabolites and eight lipid classes. Results showed that mice fed a diet enriched in saturated fatty acids had higher levels of triglycerides, cholesterol ester, monounsaturated fatty acids, lactate, 3-hydroxy-butyrate, and alanine and lower levels of glucose, compared to mice fed a control diet. In conclusion, proton NMR provided a rapid overview of the main lipid classes (triglycerides, cholesterol, phospholipids, fatty acids) and the most abundant aqueous metabolites in liver.

Mots clés: 1H-NMR spectroscopy; liver; steatosis

Mots clés: lipidomics; metabolomics quantitative profiling

Domaines: Toxicologie; Alimentation et Nutrition

Aurélien Amiel, Marie Tremblay-Franco, Roselyne Gautier, Simon Ducheix, Alexandra Montagner, et al.. Proton NMR enables the absolute quantification of aqueous metabolites and lipid classes in unique mouse liver samples. Metabolites, 2020, 10 (1), 20 p. ⟨10.3390/metabo10010009⟩. ⟨hal-02622894⟩