期刊
JOURNAL OF PROTEOME RESEARCH
卷 20, 期 6, 页码 3114-3123出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.1c00017
关键词
phosphoinositides; targeted lipidomics; ion chromatography
资金
- Deutsche Forschungsgemeinschaft
- FWF Der Wissenschaftsfonds [P33298-B]
- Federal Ministry of Education and Research (BMBF) in the German Network Bioinformatics Infrastructure (de.NBI) initiative [031L0108A, 031A534B]
- European Union [766118]
This study presents a quantitative targeted ion chromatography-mass spectrometry-based workflow for measuring phosphoinositides, which improves the accuracy of quantification. The workflow was validated in various tissues and demonstrated robustness for analyzing less-abundant phosphoinositides.
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.
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