Periodic Reporting for period 3 - SUMMIT (Site-specific Ultrasensitive Magnetic resonance of Mixtures for Isotopic Tracking)
Reporting period: 2022-10-01 to 2024-03-31
In this context, the aim of the SUMMIT project is to develop new analytical chemistry approaches in order to improve the sensitivity and resolution of NMR spectroscopy for the analysis of complex biological mixtures, and in particular for applications in metabolomics. The developments are based on a set of innovative methodological developments. These include hyperpolarization methods, relying on a prototype nuclear dynamic polarization equipment, which improves the sensitivity of NMR by several orders of magnitude. The SUMMIT project team is also developing signal detection methods based on ultrafast two-dimensional NMR spectroscopy, a method that makes it possible to better separate the signals of compounds in a mixture. Finally, we will evaluate the potential of these sensitive and rapid detection methods for a wide variety of biological issues, in fields ranging from health to agri-food.
Ultimately, the SUMMIT project has the ambition to offer a new paradigm for the analysis of complex samples by NMR spectroscopy, and to provide new analytical tools for a broad community of researchers in chemistry and biology.
On the methodological side, the main achievements so far are:
• The fine optimization of our prototype dissolution-DNP experimental setting for the robust and reproducible analysis of complex metabolic mixtures;
• The development of novel ultrafast 2D NMR experiments for the rapid screening of complex chemical samples;
• The comprehensive description of concepts underlying ultrafast 2D NMR for analytical chemistry applications;
• The development of dedicated processing tools to extract relevant data from 2D spectra of complex mixtures.
On the application side, we have demonstrated for the first time that dissolution-DNP can provide relevant information in a full metabolomics study based on 13C NMR at natural isotopic abundance. Specifically, we have demonstrated that our approach could extract biologically relevant information on plant metabolism by analyzing tomato fruit extracts at different development stages. This results paves the way to many different applications which are currently being explored, in particular through the MetaboHub national research infrastructure community in metabolomics.
-The first proof-of-concept that dissolution-DNP can provide relevant information in a full metabolomics study, opening the way to many applications in the SUMMIT project and beyond (Dey et al, Anal. Chem. 2020, 92, 14867);
-The fine optimization of a dissolution-DNP experimental setting for 13C of metabolic samples with high repeatability and sensitivity (publication in preparation)
Thanks to such advances, it is the first time that dissolution-DNP is shown to be useful for complex mixtures at natural 13C abundance. Until the end of the project, we will explore the following objectives:
-The development of tailored ultrafast 2D NMR pulse sequences to maximize the peak separation in complex hyperpolarized mixtures,
-The development of a processing package to facilitate the extraction of relevant information from 2D spectra
-The exploration of the potential of our highly sensitive and highly resolved methods for the analysis of various biological matrices such as biofluids or extracts