An efficient LC-MS method for isomer separation and detection of sugars, phosphorylated sugars, and organic acids

© Natural Resources Canada 2022. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Bibliographische Detailangaben
Veröffentlicht in:Journal of experimental botany. - 1985. - 73(2022), 9 vom: 13. Mai, Seite 2938-2952
1. Verfasser: Koley, Somnath (VerfasserIn)
Weitere Verfasser: Chu, Kevin L, Gill, Saba S, Allen, Doug K
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2022
Zugriff auf das übergeordnete Werk:Journal of experimental botany
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Central metabolism ion suppression and enhancement isomer separation isotopic labeling liquid chromatography–mass spectrometry metabolite quantification mixed-mode column chromatography mehr... oilseeds Sugars
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520 |a Assessing central carbon metabolism in plants can be challenging due to the dynamic range in pool sizes, with low levels of important phosphorylated sugars relative to more abundant sugars and organic acids. Here, we report a sensitive liquid chromatography-mass spectrometry method for analysing central metabolites on a hybrid column, where both anion-exchange and hydrophilic interaction chromatography (HILIC) ligands are embedded in the stationary phase. The liquid chromatography method was developed for enhanced selectivity of 27 central metabolites in a single run with sensitivity at femtomole levels observed for most phosphorylated sugars. The method resolved phosphorylated hexose, pentose, and triose isomers that are otherwise challenging. Compared with a standard HILIC approach, these metabolites had improved peak areas using our approach due to ion enhancement or low ion suppression in the biological sample matrix. The approach was applied to investigate metabolism in high lipid-producing tobacco leaves that exhibited increased levels of acetyl-CoA, a precursor for oil biosynthesis. The application of the method to isotopologue detection and quantification was considered through evaluating 13C-labeled seeds from Camelina sativa. The method provides a means to analyse intermediates more comprehensively in central metabolism of plant tissues 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a Research Support, U.S. Gov't, Non-P.H.S. 
650 4 |a Central metabolism 
650 4 |a ion suppression and enhancement 
650 4 |a isomer separation 
650 4 |a isotopic labeling 
650 4 |a liquid chromatography–mass spectrometry 
650 4 |a metabolite quantification 
650 4 |a mixed-mode column chromatography 
650 4 |a oilseeds 
650 7 |a Sugars  |2 NLM 
700 1 |a Chu, Kevin L  |e verfasserin  |4 aut 
700 1 |a Gill, Saba S  |e verfasserin  |4 aut 
700 1 |a Allen, Doug K  |e verfasserin  |4 aut 
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