Journal
ANALYTICAL CHEMISTRY
Volume 89, Issue 23, Pages 12857-12865Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.7b03437
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Funding
- Max Planck Gesellschaft
- Liver System Medicine (LiSyM) program - Bundesministerium fur Bildung und Forschung (BMBF)
- Lipidomics & Informatics for Life Sciences (LIES) Unit of de.NBI Consortium - Bundesministerium fur Bildung und Forschung (BMBF)
- Deutsche Forschungsgemeinschaft (DFG) [TRR83, TP17, TP18]
- German Federal Ministry of Education and Research
- European Research Council (ERC) [280271]
- Dresden International Graduate School for Biomedicine and Bioengineering, - DFG [GS97]
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Lipidomes undergo permanent extensive remodeling, but how the turnover rate differs between lipid classes and molecular species is poorly understood. We employed metabolic N-15 labeling and shotgun ultra-high-resolution mass spectrometry (sUHR) to quantify the absolute (molar) abundance and determine the turnover rate of glycerophospholipids and sphingolipids by direct analysis of total lipid extracts. sUHR performed on a commercial Orbitrap Elite instrument at the mass resolution of 1.35 x 10 (6) (m/z 200) baseline resolved peaks of C-13 isotopes of unlabeled and monoisotopic peaks of N-15 labeled lipids (Delta m = 0.0063 Da). Therefore, the rate of metabolic N-15 labeling of individual lipid species could be determined without compromising the scope, accuracy, and dynamic range of full-lipidome quantitative shotgun profiling. As a proof of concept, we employed sUHR to determine the lipidome composition and fluxes of 62 nitrogen-containing membrane lipids in human hepatoma HepG2 cells.
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