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Development of RP-UHPSFC/MS Method for Determination of Nonpolar Lipid Classes in Biological Samples

Authors: Šubrtová Veronika | Vaňková Zuzana | Peterka Ondřej | Jirásko Robert | Holčapek Michal

Year: 2023

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	Development of RP-UHPSFC/MS Method for Determination of Nonpolar Lipid Classes in Biological Samples	Lipidomics is the field related to the determination of lipids in biological systems. The term lipid includes a large group of compounds, generally soluble in organic solvents, which are chemically and structurally heterogeneous. Their groups consist of nonpolar lipids, forming important biomolecules crucial for energy production and storage in all cells. They also play an important role in cellular homeostasis, however, the importance of individual lipids in these processes is not fully understood. Their analysis is not simple. Therefore, highly effective methods that can distinguish individual lipids side by side in biological matrices are required. For this reason, lipid analyses are based on mass spectrometry (MS) either with direct infusion (shotgun) or in connection to prior separation. A very advantageous combination for the analysis of nonpolar lipids is reversed-phase ultrahigh-performance supercritical fluid chromatography (RP-UHPSFC), which can be easily combined with electrospray ionization MS. RP-UHPSFC can separate lipids into lipid classes according to their polarity and also into lipid species according to the length of the fatty acyl chains and the number and position of the double bonds. The main task of this study was to develop high-throughput RP-UHPSFC for the determination of nonpolar lipid classes, namely, fatty acids, sterols, sterol esters, mono-, di-, and triacylglycerols in plasma or serum samples. RP-UHPSFC method using sub 2 m particles columns was coupled to high-resolution mass spectrometry using a quadrupole – time of flight (QTOF) mass analyzer and electrospray ionization (ESI). Full scan and MS/MS spectra were measured in positive-ion mode and identification of individual lipids was based on retention of behavioral characteristics. In total, 389 nonpolar lipids were identified in human serum/plasma samples. This work was supported by the project No. 21-20238S sponsored by the Czech Science Foundation.