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STRUCTURAL CHARACTERIZATION OF GLYCOSPHINGOLIPIDS IN HUMAN PLASMA USING HILIC-ESI-MS/MS

Autoři: Hořejší Karel | Jirásko Robert | Kahoun David | Chocholoušková Michaela | Holčapek Michal

Rok: 2020

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

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Jazyk	Název	Abstrakt	Klíčová slova
eng	STRUCTURAL CHARACTERIZATION OF GLYCOSPHINGOLIPIDS IN HUMAN PLASMA USING HILIC-ESI-MS/MS	Glycosphingolipids (GSL) are ubiquitous membrane constituents involved in many cellular processes. These amphiphatic molecules are generally considered as one of the major and most structurally diverse subclass of sphingolipids. GSL has been shown to be implicated in the pathogenesis of various diseases, therefore, they may be used as potential diagnostic markers to differentiate patients and healthy controls based on multivariate data analysis. In this work, GSL are isolated and extracted from human plasma using a monophasic ethanol-water mixture as protein precipitation reagent. Deproteinized and reconstituted plasma extract is further purified and concentrated by C18-based solid-phase extraction. The hydrophilic interaction liquid chromatography (HILIC) coupled to an electrospray ionization tandem mass spectrometry (ESI-MS/MS) is applied for analysis of GSL in human plasma. Identification and structural elucidation of individual GSL species is based on the specific chromatographic retention in HILIC mode and comprehensive interpretation of their characteristic MS and MS/MS spectra in both polarity modes, respectively. In conclusion, we have unambiguously identified and structurally described fragmentation pathways of a total of 123 GSL species present in human plasma, which can significantly accelerate lipid identification and can be used as a valuable tool in further clinical studies.

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