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

Identifier:  imarina:6389030

Handle: https://hdl.handle.net/20.500.11797/imarina6389030

Authors:  Vinaixa, M; Rodriguez, MA; Aivio, S; Capellades, J; Gómez, J; Canyellas, N; Stracker, TH; Yanes, O

Abstract:
© 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. 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 13C-satellite peaks using 1D-1H-NMR spectra. In comparison with 13C-NMR, TOCSY and HSQC, PEPA improves sensitivity, accelerates the elucidation of 13C 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 1H-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.