Simultaneous untargeted and targeted profiling of underivatized primary metabolites on sulfur-deficient barley
Résumé
Metabolomics based-mass spectrometry are increasingly applied in diverse scientific domains, notably agronomy and plant biology, to understand plants’ behaviors under biotic/abiotic stress conditions. In fact, these stress conditions are able to disrupt many biosynthetic pathways that include mainly primary metabolites. Profiling and quantifying primary metabolites remain a challenging task because they are poorly retained in reverse phase columns due to their high polarity. The aim of our method which is to simultaneously perform an untargeted/targeted metabolite profiling in order to understand the nutrient deficiency effect on plants. Two fast and accurate methods were developed to detect and quantify amino acids, organic acids, sulfur metabolites, and secondary metabolites using an UPLC coupled to a QTOF mass spectrometer. An HSS T3 column was used to analyze amino acids and sulfur containing metabolites in positive ionization mode, and a Luna R Omega PS C18 column was used for organic acids profiling in negative mode. Ionization was achieved using an electrospray ion source (ESI). Methods were successfully applied allowing to detect, quantify and discriminate primary metabolites in short-runs without any additional sampling step such as derivatization or ion pairing. On the other hand, untargeted analysis was conducted using Progenesis QI performing alignment, peak picking, normalization, metabolite identifications and multivariate analysis. The simultaneous analysis provided cumulative information allowing to discriminate between two plant batches. Thus, discriminant biomarkers were identified and validated. A fast and innovated simultaneous untargeted/targeted method has successfully been developed and applied to sulfur deficiency on Barley.
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