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Article

Comprehensive Identification of Glycosphingolipids in Human Plasma Using Hydrophilic Interaction Liquid Chromatography—Electrospray Ionization Mass Spectrometry

1
Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 532 10 Pardubice, Czech Republic
2
Institute of Chemistry, Faculty of Science, University of South Bohemia in České Budějovice, Branišovská 1760, 370 05 České Budějovice, Czech Republic
*
Author to whom correspondence should be addressed.
Academic Editor: Leonardo Tenori
Metabolites 2021, 11(3), 140; https://doi.org/10.3390/metabo11030140
Received: 20 January 2021 / Revised: 21 February 2021 / Accepted: 23 February 2021 / Published: 26 February 2021
(This article belongs to the Special Issue Mass Spectrometry-Based Lipidomics)
Glycosphingolipids (GSL) represent a highly heterogeneous class of lipids with many cellular functions, implicated in a wide spectrum of human diseases. Their isolation, detection, and comprehensive structural analysis is a challenging task due to the structural diversity of GSL molecules. In this work, GSL subclasses are isolated from human plasma using an optimized monophasic ethanol–water solvent system capable to recover a broad range of GSL species. Obtained deproteinized plasma is subsequently purified and concentrated by C18-based solid-phase extraction (SPE). The hydrophilic interaction liquid chromatography coupled to electrospray ionization linear ion trap tandem mass spectrometry (HILIC-ESI-LIT-MS/MS) is used for GSL analysis in the human plasma extract. Our results provide an in-depth profiling and structural characterization of glycosphingolipid and some phospholipid subclasses identified in the human plasma based on their retention times and the interpretation of tandem mass spectra. The structural composition of particular lipid species is readily characterized based on the detailed interpretation of mass spectrometry (MS) and tandem mass spectrometry (MS/MS) spectra and further confirmed by specific fragmentation behavior following predictable patterns, which yields to the unambiguous identification of 154 GSL species within 7 lipid subclasses and 77 phospholipids representing the highest number of GSL species ever reported in the human plasma. The developed HILIC-ESI-MS/MS method can be used for further clinical and biological research of GSL in the human blood or other biological samples. View Full-Text
Keywords: glycosphingolipids; lipidomics; mass spectrometry; hydrophilic interaction liquid chromatography; human plasma; lipid profile; sample preparation; fragmentation behavior glycosphingolipids; lipidomics; mass spectrometry; hydrophilic interaction liquid chromatography; human plasma; lipid profile; sample preparation; fragmentation behavior
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MDPI and ACS Style

Hořejší, K.; Jirásko, R.; Chocholoušková, M.; Wolrab, D.; Kahoun, D.; Holčapek, M. Comprehensive Identification of Glycosphingolipids in Human Plasma Using Hydrophilic Interaction Liquid Chromatography—Electrospray Ionization Mass Spectrometry. Metabolites 2021, 11, 140. https://doi.org/10.3390/metabo11030140

AMA Style

Hořejší K, Jirásko R, Chocholoušková M, Wolrab D, Kahoun D, Holčapek M. Comprehensive Identification of Glycosphingolipids in Human Plasma Using Hydrophilic Interaction Liquid Chromatography—Electrospray Ionization Mass Spectrometry. Metabolites. 2021; 11(3):140. https://doi.org/10.3390/metabo11030140

Chicago/Turabian Style

Hořejší, Karel, Robert Jirásko, Michaela Chocholoušková, Denise Wolrab, David Kahoun, and Michal Holčapek. 2021. "Comprehensive Identification of Glycosphingolipids in Human Plasma Using Hydrophilic Interaction Liquid Chromatography—Electrospray Ionization Mass Spectrometry" Metabolites 11, no. 3: 140. https://doi.org/10.3390/metabo11030140

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