1. Positional Enrichment by Proton Analysis (PEPA): A One‐Dimensional 1 H‐NMR Approach for 13 C Stable Isotope Tracer Studies in Metabolomics
- Author
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Josep Gómez, Maria Vinaixa, Suvi Aivio, Nicolau Canyellas, Jordi Capellades, Miguel A. Rodríguez, Oscar Yanes, and Travis H. Stracker
- Subjects
0301 basic medicine ,PEPA ,Chromatography ,Chemistry ,Stable isotope ratio ,General Medicine ,General Chemistry ,010402 general chemistry ,01 natural sciences ,Catalysis ,0104 chemical sciences ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Metabolomics ,TRACER ,Proton NMR ,030217 neurology & neurosurgery ,Heteronuclear single quantum coherence spectroscopy ,Human cancer - Abstract
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 13 C-satellite peaks using 1D-1 H-NMR spectra. In comparison with 13 C-NMR, TOCSY and HSQC, PEPA improves sensitivity, accelerates the elucidation of 13 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 1 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.
- Published
- 2017