Pure Shift Nuclear Magnetic Resonance: a New Tool for Plant Metabolomics

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Baena-Moncada, Angelica M.
Calderon Gomez, J. C.
Goncalves, Josue M.
Quispe-Garrido, Lady V.
Ruiz-Montoya, Jose G.
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Journal of Visualized Experiments
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Nuclear Magnetic Resonance (NMR) is one of the most powerful tools used in metabolomics. It stands as a highly accurate and reproducible method that not only provides quantitative data but also permits structural identification of the metabolites present in complex mixtures. Metabolic profiling by H-1 NMR has proven useful in the study of various types of plant scenarios, which include the evaluation of crop conditions, harvest and postharvest treatments, metabolic phenotyping, metabolic pathways, gene regulation, identification of biomarkers, chemotaxonomy, quality control, denomination of origin, among others. However, signal overlapping of the large number of resonances with expanded J-coupling multiplicities complicates the spectra analysis and its interpretation, and represents a limitation for classical H-1 NMR profiling. In the last decade, novel NMR broadband homonuclear decoupling techniques through which multiplet signals collapse into single resonance lines - commonly called Pure Shift methods - have been developed to overcome the spectra resolution problem inherent to H-1 NMR classical spectra. Here a step-by-step protocol of the plant extract preparation and the procedure to record optimal Pure Shift PSYCHE and SAPPHIRE-PSYCHE spectra in three different plant matrices - Vanilla plant leaves, potato tubers (S. tuberosum), and Cape gooseberries (P. peruviana) - is presented. The effect of the gain in resolution in metabolic identification, correlation analysis and multivariate analyses, as compared against classical spectra, is discussed.
This study was funded by the Consejo Nacional de Ciencia, Tecnologia e Innovacion Tecnologica (CONCYTEC) -Programa Atraccion de Investigadores Cienciactiva Contract #008-2017-FONDECYT.
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plant scenarios