Abstract
Drought is a major limiting factor in agriculture and responsible for dramatic crop yield losses worldwide. The adjustment of the metabolic status via accumulation of drought stress-responsive osmolytes is one of the many strategies that some plants have developed to cope with water deficit conditions. Osmolytes are highly polar compounds, analysis of whcih is difficult with typical reversed-phase chromatography. Porous graphitic carbon (PGC) has shown to be a suitable alternative to reversed-phase stationary phases for the analysis of highly polar compounds typically found in the plant metabolome. In this chapter, we describe the development and validation of a PGC-based liquid chromatography tandem mass spectrometry (LC-MSn) method suitable for the target analysis of water-soluble carbohydrates, such as raffinose family oligosaccharides (RFOs). We present detailed information regarding PGC column equilibration, LC-MSn system operation, data analysis, and important notes to be considered during the steps of method development and validation.
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Acknowledgments
This work was supported by the FCT Investigator Programme (IF/00376/2012/CP0165/CT0003) from Fundação para a Ciência e a Tecnologia and the ITQB NOVA research unit GREEN-it “Bioresources for sustainability” (UID/Multi/04551/2013). TFJ acknowledges FCT for the PhD grant (PD/BD/113475/2015) from the ITQB PhD programme “Plants for Life” (PD/00035/2013).
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Jorge, T.F., Florêncio, M.H., António, C. (2017). Porous Graphitic Carbon Liquid Chromatography–Mass Spectrometry Analysis of Drought Stress-Responsive Raffinose Family Oligosaccharides in Plant Tissues. In: Sunkar, R. (eds) Plant Stress Tolerance. Methods in Molecular Biology, vol 1631. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7136-7_17
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DOI: https://doi.org/10.1007/978-1-4939-7136-7_17
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