Positional Enrichment by Proton Analysis (PEPA): A One-Dimensional 1 H-NMR Approach for 13 C Stable Isotope Tracer Studies in Metabolomics

A novel metabolomics approach for NMR-based stable isotope tracer studies called PEPA is presented, and its performance validated using human cancer cells. PEPA detects the position of carbon label in isotopically enriched metabolites and quantifies fractional enrichment by indirect determination of...

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Bibliographic Details
Published in:Angewandte Chemie International Edition 2017-03, Vol.56 (13), p.3531-3535
Main Authors: Vinaixa, Maria, Rodríguez, Miguel A, Aivio, Suvi, Capellades, Jordi, Gómez, Josep, Canyellas, Nicolau, Stracker, Travis H, Yanes, Oscar
Format: Article
Language:English
Subjects:
Carbon Isotopes - analysis
Cell Line, Tumor
High-Throughput Screening Assays - methods
Humans
Metabolome
Metabolomics - methods
Proton Magnetic Resonance Spectroscopy - methods
Protons
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Summary:A novel metabolomics approach for NMR-based stable isotope tracer studies called PEPA is presented, and its performance validated using human cancer cells. PEPA detects the position of carbon label in isotopically enriched metabolites and quantifies fractional enrichment by indirect determination of C-satellite peaks using 1D- H-NMR spectra. In comparison with C-NMR, TOCSY and HSQC, PEPA improves sensitivity, accelerates the elucidation of C positions in labeled metabolites and the quantification of the percentage of stable isotope enrichment. Altogether, PEPA provides a novel framework for extending the high-throughput of H-NMR metabolic profiling to stable isotope tracing in metabolomics, facilitating and complementing the information derived from 2D-NMR experiments and expanding the range of isotopically enriched metabolites detected in cellular extracts.
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.201611347