Profiling Intact Glycosphingolipids with Automated Structural Annotation and Quantitation from Human Samples with Nanoflow Liquid Chromatography Mass Spectrometry

Sphingolipids are an essential subset of bioactive lipids found in most eukaryotic cells that contribute to membrane biophysical properties and are involved in cellular differentiation, recognition, and mediating interactions. The described nanoHPLC-ESI-Q/ToF methodology utilizes known biosynthetic...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2024-04, Vol.96 (15), p.5951-5959
Main Authors: Schindler, Ryan L., Oloumi, Armin, Tena, Jennyfer, Alvarez, Michael Russelle S., Liu, Yiyun, Grijaldo, Sheryl, Barboza, Mariana, Jin, Lee-Way, Zivkovic, Angela M., Lebrilla, Carlito B.
Format: Article
Language:English
Subjects:
Annotations
Biomolecules
Chromatography
Chromatography, High Pressure Liquid - methods
Chromatography, Liquid - methods
Differentiation (biology)
Flow rates
Glycosphingolipids
Human tissues
Humans
Lipids
Liquid chromatography
Liquid Chromatography-Mass Spectrometry
Mass spectrometry
Mass spectroscopy
Profiling
Reproducibility of Results
Robustness
Sphingolipids
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Summary:Sphingolipids are an essential subset of bioactive lipids found in most eukaryotic cells that contribute to membrane biophysical properties and are involved in cellular differentiation, recognition, and mediating interactions. The described nanoHPLC-ESI-Q/ToF methodology utilizes known biosynthetic pathways, accurate mass detection, optimized collision-induced disassociation, and a robust nanoflow chromatographic separation for the analysis of intact sphingolipids found in human tissue, cells, and serum. The methodology was developed and validated with an emphasis on addressing the common issues experienced in profiling these amphipathic lipids, which are part of the glycocalyx and lipidome. The high sensitivity obtained using nanorange flow rates with robust chromatographic reproducibility over a wide range of concentrations and injection volumes results in confident identifications for profiling these low-abundant biomolecules.
ISSN:0003-2700
1520-6882
1520-6882
DOI:10.1021/acs.analchem.4c00077