Quantification of sugars and sugar phosphates in Arabidopsis thaliana tissues using porous graphitic carbon liquid chromatography-electrospray ionization mass spectrometry

Carla Antonio; Tony Larson; Alison Gilday; Ian Graham; Ed Bergstroem; Jane Thomas-Oates

doi:10.1016/j.chroma.2007.10.011

Quantification of sugars and sugar phosphates in Arabidopsis thaliana tissues using porous graphitic carbon liquid chromatography-electrospray ionization mass spectrometry

Carla Antonio, Tony Larson, Alison Gilday, Ian Graham, Ed Bergstroem, Jane Thomas-Oates

Research output: Contribution to journal › Article › peer-review

Abstract

This work reports the development and optimisation of a negative ion mode on-line LC-ESI-MS/MS method for the sensitive targeted analysis of the key glycolytic intermediates, sugars and sugar phosphates from plants, using a porous graphitic carbon (PGC) stationary phase and an MS compatible mobile phase. Using this newly developed method, separation and detection of a solution of standard compounds is achieved in less than 20 min. Target metabolite compounds were identified in plant extracts from their characteristic retention times, and product ion spectra. This on-line PGC-ESI-MS/MS method shows good linearity over the concentration range 0-100 LM, selectivity, short analysis time, and limits of detection of 0. 1 LM for disaccharides trehalose (Tre), sucrose (Suc), and maltose, and 1.5 lLM for hexose phosphates fructose-6-phosphate (Fru6P), glucose- I -phosphate (GIcIP), and glucose-6-phosphate (Glc6P), and phosphoenolpyruvate, (PEP). This paper describes details of our method and its application to the simultaneous quantitative analysis of soluble sugars and sugar phosphates from Arabidopsis thaliana tissues. We have demonstrated the utility of our method for the analysis of biological samples by applying it to the simultaneous quantitation of changes in soluble sugars and sugar phosphates in A. thaliana Columbia-0 (Col-0) and its starchless phosphoglucomutase (pgm) mutant over a 12-h light/I 2-h dark growth cycle. (c) 2007 Elsevier B.V. All rights reserved.

Original language	English
Pages (from-to)	170-178
Number of pages	9
Journal	Journal of Chromatography A
Volume	1172
Issue number	2
DOIs	https://doi.org/10.1016/j.chroma.2007.10.011
Publication status	Published - 23 Nov 2007

Keywords

glycolytic intermediates
sugar phosphates
porous graphitic carbon
HPLC-MS/MS
Arabidopsis thaliana
pgmI mutant
ANION-EXCHANGE CHROMATOGRAPHY
PULSED AMPEROMETRIC DETECTION
EVAPORATIVE LIGHT-SCATTERING
ESCHERICHIA-COLI
STATIONARY-PHASE
INORGANIC PYROPHOSPHATE
ION SUPPRESSION
POLAR COMPOUNDS
PERFORMANCE
METABOLITES

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10.1016/j.chroma.2007.10.011

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@article{37a7e91a8884475084e01ac5dd108048,

title = "Quantification of sugars and sugar phosphates in Arabidopsis thaliana tissues using porous graphitic carbon liquid chromatography-electrospray ionization mass spectrometry",

abstract = "This work reports the development and optimisation of a negative ion mode on-line LC-ESI-MS/MS method for the sensitive targeted analysis of the key glycolytic intermediates, sugars and sugar phosphates from plants, using a porous graphitic carbon (PGC) stationary phase and an MS compatible mobile phase. Using this newly developed method, separation and detection of a solution of standard compounds is achieved in less than 20 min. Target metabolite compounds were identified in plant extracts from their characteristic retention times, and product ion spectra. This on-line PGC-ESI-MS/MS method shows good linearity over the concentration range 0-100 LM, selectivity, short analysis time, and limits of detection of 0. 1 LM for disaccharides trehalose (Tre), sucrose (Suc), and maltose, and 1.5 lLM for hexose phosphates fructose-6-phosphate (Fru6P), glucose- I -phosphate (GIcIP), and glucose-6-phosphate (Glc6P), and phosphoenolpyruvate, (PEP). This paper describes details of our method and its application to the simultaneous quantitative analysis of soluble sugars and sugar phosphates from Arabidopsis thaliana tissues. We have demonstrated the utility of our method for the analysis of biological samples by applying it to the simultaneous quantitation of changes in soluble sugars and sugar phosphates in A. thaliana Columbia-0 (Col-0) and its starchless phosphoglucomutase (pgm) mutant over a 12-h light/I 2-h dark growth cycle. (c) 2007 Elsevier B.V. All rights reserved.",

keywords = "glycolytic intermediates, sugar phosphates, porous graphitic carbon, HPLC-MS/MS, Arabidopsis thaliana, pgmI mutant, ANION-EXCHANGE CHROMATOGRAPHY, PULSED AMPEROMETRIC DETECTION, EVAPORATIVE LIGHT-SCATTERING, ESCHERICHIA-COLI, STATIONARY-PHASE, INORGANIC PYROPHOSPHATE, ION SUPPRESSION, POLAR COMPOUNDS, PERFORMANCE, METABOLITES",

author = "Carla Antonio and Tony Larson and Alison Gilday and Ian Graham and Ed Bergstroem and Jane Thomas-Oates",

year = "2007",

month = nov,

day = "23",

doi = "10.1016/j.chroma.2007.10.011",

language = "English",

volume = "1172",

pages = "170--178",

journal = "Journal of Chromatography A",

issn = "0021-9673",

publisher = "Elsevier",

number = "2",

}

TY - JOUR

T1 - Quantification of sugars and sugar phosphates in Arabidopsis thaliana tissues using porous graphitic carbon liquid chromatography-electrospray ionization mass spectrometry

AU - Antonio, Carla

AU - Larson, Tony

AU - Gilday, Alison

AU - Graham, Ian

AU - Bergstroem, Ed

AU - Thomas-Oates, Jane

PY - 2007/11/23

Y1 - 2007/11/23

N2 - This work reports the development and optimisation of a negative ion mode on-line LC-ESI-MS/MS method for the sensitive targeted analysis of the key glycolytic intermediates, sugars and sugar phosphates from plants, using a porous graphitic carbon (PGC) stationary phase and an MS compatible mobile phase. Using this newly developed method, separation and detection of a solution of standard compounds is achieved in less than 20 min. Target metabolite compounds were identified in plant extracts from their characteristic retention times, and product ion spectra. This on-line PGC-ESI-MS/MS method shows good linearity over the concentration range 0-100 LM, selectivity, short analysis time, and limits of detection of 0. 1 LM for disaccharides trehalose (Tre), sucrose (Suc), and maltose, and 1.5 lLM for hexose phosphates fructose-6-phosphate (Fru6P), glucose- I -phosphate (GIcIP), and glucose-6-phosphate (Glc6P), and phosphoenolpyruvate, (PEP). This paper describes details of our method and its application to the simultaneous quantitative analysis of soluble sugars and sugar phosphates from Arabidopsis thaliana tissues. We have demonstrated the utility of our method for the analysis of biological samples by applying it to the simultaneous quantitation of changes in soluble sugars and sugar phosphates in A. thaliana Columbia-0 (Col-0) and its starchless phosphoglucomutase (pgm) mutant over a 12-h light/I 2-h dark growth cycle. (c) 2007 Elsevier B.V. All rights reserved.

AB - This work reports the development and optimisation of a negative ion mode on-line LC-ESI-MS/MS method for the sensitive targeted analysis of the key glycolytic intermediates, sugars and sugar phosphates from plants, using a porous graphitic carbon (PGC) stationary phase and an MS compatible mobile phase. Using this newly developed method, separation and detection of a solution of standard compounds is achieved in less than 20 min. Target metabolite compounds were identified in plant extracts from their characteristic retention times, and product ion spectra. This on-line PGC-ESI-MS/MS method shows good linearity over the concentration range 0-100 LM, selectivity, short analysis time, and limits of detection of 0. 1 LM for disaccharides trehalose (Tre), sucrose (Suc), and maltose, and 1.5 lLM for hexose phosphates fructose-6-phosphate (Fru6P), glucose- I -phosphate (GIcIP), and glucose-6-phosphate (Glc6P), and phosphoenolpyruvate, (PEP). This paper describes details of our method and its application to the simultaneous quantitative analysis of soluble sugars and sugar phosphates from Arabidopsis thaliana tissues. We have demonstrated the utility of our method for the analysis of biological samples by applying it to the simultaneous quantitation of changes in soluble sugars and sugar phosphates in A. thaliana Columbia-0 (Col-0) and its starchless phosphoglucomutase (pgm) mutant over a 12-h light/I 2-h dark growth cycle. (c) 2007 Elsevier B.V. All rights reserved.

KW - glycolytic intermediates

KW - sugar phosphates

KW - porous graphitic carbon

KW - HPLC-MS/MS

KW - Arabidopsis thaliana

KW - pgmI mutant

KW - ANION-EXCHANGE CHROMATOGRAPHY

KW - PULSED AMPEROMETRIC DETECTION

KW - EVAPORATIVE LIGHT-SCATTERING

KW - ESCHERICHIA-COLI

KW - STATIONARY-PHASE

KW - INORGANIC PYROPHOSPHATE

KW - ION SUPPRESSION

KW - POLAR COMPOUNDS

KW - PERFORMANCE

KW - METABOLITES

U2 - 10.1016/j.chroma.2007.10.011

DO - 10.1016/j.chroma.2007.10.011

M3 - Article

SN - 0021-9673

VL - 1172

SP - 170

EP - 178

JO - Journal of Chromatography A

JF - Journal of Chromatography A

IS - 2

ER -