Targeted Phosphoinositides Analysis Using High-Performance Ion Chromatography-Coupled Selected Reaction Monitoring Mass Spectrometry.
Yam Fung Hilaire CheungCristina ComanPhilipp WesthoffMailin MankeAlbert SickmannOliver BorstMeinrad GawazSteve P WatsonJohan W M HeemskerkRobert AhrendsPublished in: Journal of proteome research (2021)
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.
Keyphrases
- mass spectrometry
- liquid chromatography
- high resolution
- tandem mass spectrometry
- high performance liquid chromatography
- electronic health record
- gas chromatography
- high speed
- capillary electrophoresis
- cancer therapy
- adipose tissue
- heart rate
- atomic force microscopy
- oxidative stress
- resting state
- solid phase extraction
- functional connectivity
- white matter
- blood pressure
- heart rate variability
- cell cycle arrest
- cerebral ischemia
- drug delivery
- cell proliferation
- brain injury
- single molecule
- subarachnoid hemorrhage
- red blood cell