NMR Analysis of Carboxylate Isotopomers of ¹³C-Metabolites by Chemoselective Derivatization with ¹⁵N-Cholamine.

ABSTRACT: A substantial fraction of common metabolites contains carboxyl functional groups. Their ¹³C isotopomer analysis by nuclear magnetic resonance (NMR) is hampered by the low sensitivity of the ¹³C nucleus, the slow longitudinal relaxation for the lack of an attached proton, and the relatively low chemical shift dispersion of carboxylates. Chemoselective (CS) derivatization is a means of tagging compounds in a complex mixture via a specific functional group. ¹⁵N₁-cholamine has been shown to be a useful CS agent for carboxylates, producing a peptide bond that can be detected via ¹⁵N-attached H with high sensitivity in heteronuclear single quantum coherence experiments. Here, we report an improved method of derivatization and show how ¹³C-enrichment at the carboxylate and/or the adjacent carbon can be determined via one- and two-bond coupling of the carbons adjacent to the cholamine ¹⁵N atom in the derivatives. We have applied this method for the determination of ¹³C isotopomer distribution in the extracts of A549 cell culture and liver tissue from a patient-derived xenograft mouse.

SUBMITTER: Vicente-Munoz S 

PROVIDER: S-EPMC9350721 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature