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Hydrophilic interaction chromatography/electrospray mass spectrometry analysis of carbohydrate-related metabolites from Arabidopsis thaliana leaf tissue

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JournalRapid Communications in Mass Spectrometry
DatePublished - May 2008
Issue number9
Volume22
Number of pages9
Pages (from-to)1399-1407
Original languageEnglish

Abstract

This work describes the development and application of an on-line liquid chromatography/mass spectrometry (LC/MS) method using hydrophilic interaction chromatography (HILIC) coupled to negative ion mode electrospray ionisation ion trap mass spectrometry (ESI-MS) for the analysis of highly polar carbohydrate-related metabolites commonly found in plants, ranging from reducing and non-reducing sugars and sugar alcohols to sugar phosphates. Using this method, separation and detection of a mixture of eight authentic standard compounds containing glucose (Glc), sucrose (Suc), raffinose, verbascose, mannitol, maltitol, glucose-6-phosphate (GIc6P) and trehalose-6-phosphate (Tre6P) were achieved in less than 15 min. The method is rapid, robust, selective, and sensitive, with limits of detection (LODs) ranging from 0.2 mu M obtained for neutral sugars, to 1.0 mu M obtained for sugar alcohols, and 2.0 mu M obtained for negatively charged sugar phosphates. We have studied the negative ion collision-induced dissociation (CID) fragmentation behaviour of the non-reducing raffinose family oligosaccharides (RFOs) raffinose, stachyose, and verbascose. Mainly B-i and C-i glycosidic and A(i) cross-ring structurally informative cleavages are observed. We have applied this HILIC/ESI-MS method for the analysis of Arabidopsis thaliana wild-type Columbia-0 (Col-0) and its starchless phosphoglucomutase mutant (pgm1) leaf extracts. The method was used to quantify Glc, Suc, raffinose, and Glc6P in A. thatiana extracts. Data obtained using this HILIC/ESI-MS method were compared with those obtained using a comparable porous graphitic carbon-based LC/ESI-MS method. Copyright (c) 2008 John Wiley & Sons, Ltd.

    Research areas

  • PERFORMANCE LIQUID-CHROMATOGRAPHY, ANION-EXCHANGE CHROMATOGRAPHY, POROUS GRAPHITIC CARBON, SMALL POLAR COMPOUNDS, FUNCTIONAL GENOMICS, GAS-CHROMATOGRAPHY, STATIONARY-PHASE, ION SUPPRESSION, OLIGOSACCHARIDES, METABOLOMICS

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