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

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 chemi...

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Published inAnalytical chemistry (Washington) Vol. 93; no. 17; pp. 6629 - 6637
Main Authors Vicente-Muñoz, Sara, Lin, Penghui, Fan, Teresa W.-M, Lane, Andrew N
Format Journal Article
LanguageEnglish
Published Washington American Chemical Society 04.05.2021
Subjects
Carbon 13
Carboxylates
Cell culture
Chemical equilibrium
Chemistry
Coherence
Functional groups
Hepatocytes
Metabolites
Nitrogen isotopes
NMR
Nuclear magnetic resonance
Quantum phenomena
Sensitivity
Tissue culture
Xenografts
Xenotransplantation
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Summary: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.
Bibliography:SVM: NMR-Based Metabolomics Core Facility, Division of Pathology and Laboratory Medicine, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, OH 45229, USA
SVM and PL carried out the derivatizations, recorded NMR spectra, analyzed data and co-wrote the manuscript; TWMF conceived and contributed to the experimental design and co-wrote the manuscript; ANL conceived and supervised the project, interpreted results and co-wrote the manuscript.
Present address
These authors contributed equally to this article.
Author Contributions
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.0c04220