Quantification of low-abundant phosphorylated carbohydrates using HILIC-QqQ-MS/MS

Tiago F. Jorge, Carla António

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Phosphorylated carbohydrates are central metabolites involved in key plant metabolic pathways, such as glycolysis and central carbon metabolism. Such pathways influence plant growth, development, and stress responses to environmental changes, and ultimately, reflect the plant’s energy status. The high polarity of these metabolites, the variety of isomeric structures (e.g., glucose-1-phosphate (G1P)/fructose-6-phosphate (F6P)/mannose-6-phosphate (M6P)/G6P, sucrose-6-phosphate (S6P)/T6P), and rapid metabolic turnover makes their analysis particularly challenging. In this chapter, we describe the use of a set of known phosphorylated carbohydrates to develop and validate a hydrophilic interaction chromatography (HILIC) triple quadrupole (QqQ) tandem mass spectrometry (MS/MS) method in the highly sensitive and selective multiple reaction monitoring (MRM) mode for the target analysis of G1P, F6P, M6P, G6P, S6P, T6P, and the sugar nucleotide uridine 5-diphospho-glucose (UDPG). We present detailed information regarding HILIC column chemistry and practical considerations when coupling it with a QqQ-MS system.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages71-86
Number of pages16
Volume1778
DOIs
Publication statusPublished - 1 Jan 2018

Publication series

NameMethods in Molecular Biology
Volume1778
ISSN (Print)1064-3745

Keywords

  • HILIC
  • Metabolite quantification
  • Method development
  • Phosphorylated carbohydrates
  • Trehalose-6-phosphate
  • Triple quadrupole

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    Jorge, T. F., & António, C. (2018). Quantification of low-abundant phosphorylated carbohydrates using HILIC-QqQ-MS/MS. In Methods in Molecular Biology (Vol. 1778, pp. 71-86). (Methods in Molecular Biology; Vol. 1778). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7819-9_6