Targeted Phosphoinositides Analysis Using High-Performance Ion Chromatography-Coupled Selected Reaction Monitoring Mass Spectrometry

Hilaire Yam Fung Cheung, Cristina Coman, Philipp Westhoff, Mailin Manke, Albert Sickmann, Oliver Borst, Meinrad Gawaz, Steve P. Watson, Johan W. M. Heemskerk, Robert Ahrends*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

4 Citations (Web of Science)

Abstract

Phosphoinositides are minor components of cell membranes, but play crucial roles in numerous signal transduction pathways. To obtain quantitative measures of phosphoinositides, sensitive, accurate, and comprehensive methods are needed. Here, we present a quantitative targeted ion chromatography-mass spectrometry-based workflow that separates phosphoinositide isomers and increases the quantitative accuracy of measured phosphoinositides. Besides testing different analytical characteristics such as extraction and separation efficiency, the reproducibility of the developed workflow was also investigated. The workflow was verified in resting and stimulated human platelets, fat cells, and rat hippocampal brain tissue, where the LOD and LOQ for phosphoinositides were at 312.5 and 625 fmol, respectively. The robustness of the workflow is shown with different applications that confirms its suitability to analyze multiple less-abundant phosphoinositides.

Original languageEnglish
Pages (from-to)3114-3123
Number of pages10
JournalJournal of Proteome Research
Volume20
Issue number6
DOIs
Publication statusPublished - 4 Jun 2021

Keywords

  • phosphoinositides
  • targeted lipidomics
  • ion chromatography
  • NECK-CANCER CELLS
  • RAT-BRAIN
  • INSULIN
  • QUANTIFICATION
  • LIPIDS
  • METABOLITES
  • 3-KINASE
  • HEAD

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