title: Ultrafast quantitative 2D NMR: an efficient tool for the measurement of specific isotopic enrichments in complex biological mixtures.
abstract: Two-dimensional nuclear magnetic resonance (2D NMR) is a promising tool for studying metabolic fluxes by measuring (13)C-enrichments in complex mixtures of (13)C-labeled metabolites. However, the methods reported so far are hampered by very long acquisition durations limiting the use of 2D NMR as a quantitative tool for fluxomics. In this paper, we propose a new approach for measuring specific (13)C-enrichments in a very fast way, by using new experiments based on ultrafast 2D NMR. Two homonuclear 2D experiments (ultrafast COSY and zTOCSY) are proposed to measure (13)C-enrichments in a single scan. Their advantages and limitations are discussed, and their high analytical potentialities are highlighted. Both methods are characterized by an accuracy of 1-2%, an average precision of 3%, and an excellent linearity. The analytical performance is equivalent or better than any of the conventional methods previously reported. The two ultrafast experiments are applied to the measurement of (13)C-enrichments on a biomass hydrolyzate, showing the first known application of ultrafast 2D NMR to a real biological extract. The experiment duration is divided by 200 compared to the conventional methods, while preserving 80% of the quantitative information. This new approach opens new perspectives of application for fluxomics and metabonomics.
authors.affiliation: Université de Nantes, CNRS, CEISAM UMR 6230, B.P. 92208, 2 rue de la Houssinière, 44322 Nantes Cedex 03, France. patrick.giraudeau@univ-nantes.fr
authors.forename: Patrick
authors.initials: P
authors.lastname: Giraudeau
authors.forename: Stéphane
authors.initials: S
authors.lastname: Massou
authors.forename: Yoann
authors.initials: Y
authors.lastname: Robin
authors.forename: Edern
authors.initials: E
authors.lastname: Cahoreau
authors.forename: Jean-Charles
authors.initials: JC
authors.lastname: Portais
authors.forename: Serge
authors.initials: S
authors.lastname: Akoka
mesh_terms: D018533:Biomass; D002247:Carbon Isotopes / Q000032:analysis; D004926:Escherichia coli / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011492:Protein Hydrolysates / Q000737:chemistry*
chemical_list: D002247:Carbon Isotopes; D011492:Protein Hydrolysates