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Metabolites
Volume 10
Issue 9
10.3390/metabo10090338
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Addendum published on 30 November 2020, see Metabolites 2020, 10(12), 490.
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Article

An Optimized Dual Extraction Method for the Simultaneous and Accurate Analysis of Polar Metabolites and Lipids Carried out on Single Biological Samples

by
Joran Villaret-Cazadamont
1,
Nathalie Poupin
1,
Anthony Tournadre
2,3,
Aurélie Batut
2,3,
Lara Gales
3,4,
Daniel Zalko
1,
Nicolas J. Cabaton
1,
Floriant Bellvert
3,4,*,† and
Justine Bertrand-Michel
2,3,*,†
1
Toxalim (Research Centre in Food Toxicology), Université de Toulouse, INRAE, ENVT, INP-Purpan, UPS, 31027 Toulouse, France
2
MetaboHUB-MetaToul-Lipidomics Core Facility, Inserm U1048, 31432 Toulouse, France
3
MetaboHUB-MetaToul, National Infrastructure for Metabolomics and Fluxomics, 31077 Toulouse, France
4
Toulouse Biotechnology Institute, Université de Toulouse, CNRS, INRAE, INSA, 31400 Toulouse, France
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Metabolites 2020, 10(9), 338; https://doi.org/10.3390/metabo10090338
Submission received: 24 July 2020 / Revised: 16 August 2020 / Accepted: 17 August 2020 / Published: 19 August 2020
(This article belongs to the Section Integrative Metabolomics)
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Abstract

The functional understanding of metabolic changes requires both a significant investigation into metabolic pathways, as enabled by global metabolomics and lipidomics approaches, and the comprehensive and accurate exploration of specific key pathways. To answer this pivotal challenge, we propose an optimized approach, which combines an efficient sample preparation, aiming to reduce the variability, with a biphasic extraction method, where both the aqueous and organic phases of the same sample are used for mass spectrometry analyses. We demonstrated that this double extraction protocol allows working with one single sample without decreasing the metabolome and lipidome coverage. It enables the targeted analysis of 40 polar metabolites and 82 lipids, together with the absolute quantification of 32 polar metabolites, providing comprehensive coverage and quantitative measurement of the metabolites involved in central carbon energy pathways. With this method, we evidenced modulations of several lipids, amino acids, and energy metabolites in HepaRG cells exposed to fenofibrate, a model hepatic toxicant, and metabolic modulator. This new protocol is particularly relevant for experiments involving limited amounts of biological material and for functional metabolic explorations and is thus of particular interest for studies aiming to decipher the effects and modes of action of metabolic disrupting compounds.
Keywords: lipidomics; metabolomics; multi-omics analysis; sample preparation; dual extraction; hepatotoxicity lipidomics; metabolomics; multi-omics analysis; sample preparation; dual extraction; hepatotoxicity

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MDPI and ACS Style

Villaret-Cazadamont, J.; Poupin, N.; Tournadre, A.; Batut, A.; Gales, L.; Zalko, D.; Cabaton, N.J.; Bellvert, F.; Bertrand-Michel, J. An Optimized Dual Extraction Method for the Simultaneous and Accurate Analysis of Polar Metabolites and Lipids Carried out on Single Biological Samples. Metabolites 2020, 10, 338. https://doi.org/10.3390/metabo10090338

AMA Style

Villaret-Cazadamont J, Poupin N, Tournadre A, Batut A, Gales L, Zalko D, Cabaton NJ, Bellvert F, Bertrand-Michel J. An Optimized Dual Extraction Method for the Simultaneous and Accurate Analysis of Polar Metabolites and Lipids Carried out on Single Biological Samples. Metabolites. 2020; 10(9):338. https://doi.org/10.3390/metabo10090338

Chicago/Turabian Style

Villaret-Cazadamont, Joran, Nathalie Poupin, Anthony Tournadre, Aurélie Batut, Lara Gales, Daniel Zalko, Nicolas J. Cabaton, Floriant Bellvert, and Justine Bertrand-Michel. 2020. "An Optimized Dual Extraction Method for the Simultaneous and Accurate Analysis of Polar Metabolites and Lipids Carried out on Single Biological Samples" Metabolites 10, no. 9: 338. https://doi.org/10.3390/metabo10090338

APA Style

Villaret-Cazadamont, J., Poupin, N., Tournadre, A., Batut, A., Gales, L., Zalko, D., Cabaton, N. J., Bellvert, F., & Bertrand-Michel, J. (2020). An Optimized Dual Extraction Method for the Simultaneous and Accurate Analysis of Polar Metabolites and Lipids Carried out on Single Biological Samples. Metabolites, 10(9), 338. https://doi.org/10.3390/metabo10090338

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