RMN 13 C hyperpolarisée pour la métabolomique

Hyperpolarized 13 C NMR relying on dissolution dynamic nuclear polarization (d-DNP) is able to provide rich and highly resolved spectral fingerprint at natural abundance and in a repeatable way. Such 13 C NMR spectra with sub-mM limit of detection represents an opportunity for complex mixture analysis and has already been exploited in metabolomic context with concentrated plant extracts. Nevertheless, the gain of sensitivity brought by d-DNP comes with new challenges that currently prevent general use of the method in omics sciences. In this context, this thesis work focuses on the exploration of its potential for complex mixture analysis and omics sciences. For instance, we showed that d-DNP 13 C NMR is also able to provide rich spectra on a dilute biofluid, such as urine. Moreover, being able to recover unique quantitative information using standard additions on urinary metabolites e that were inaccessible with conventional 1 H NMR due to strong peak overlap. We reported the first application of this emerging hyperpolarized approach to a metabolomics study involving clinical samples. The analysis of human urine samples from patients with different stage of chronic kidney disease was performed using both hyperpolarized 13 C NMR and benchmarked conventional 1 H NMR at high field. The results highlighted the complementarity of biomarker coverage between the two methods, in particular, 13 C d- DNP NMR allowed the annotation of two biomarkers – guanine and guanidoacetate – that could not be detected by 1 H NMR because of peak overlap.