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Comprehensive Analysis of Nonpolar Lipids in Human Plasma and Serum Samples using RP-UHPSFC/MS

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

Year: 2024

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	Comprehensive Analysis of Nonpolar Lipids in Human Plasma and Serum Samples using RP-UHPSFC/MS	Supercritical fluid chromatography (SFC) is gaining popularity as a separation technique in lipidomics due to its hybrid nature, which combines the advantageous properties of gas and liquid chromatography. SFC techniques provide high sensitivity for analysis of low-polar compounds, such as nonpolar lipids, due to supercritical carbon dioxide used as mobile phase. The combination of sub-2-µm particles as stationary phase and the low viscosity of the mobile phase enables high separation efficiency and shorter analysis times compared to liquid chromatography. The connection of mass spectrometry (MS) and ultrahigh-performance supercritical fluid chromatography (UHPSFC) makes the method ideal for lipidomic analysis. SFC allows to use two separation approaches for lipidomic analysis. Lipid class separation (HILIC-like mode) separates lipids according to polar head group, resulting in coelution of lipids from one lipid class in one chromatographic peak, which is preferred for quantitative analysis. Otherwise, the lipid species separation approach (RP-like mode) separates lipids based on the length of the fatty acyl chain and the number of double bonds enabling the separation of isomers with different number and positions of the double bond. The main goal of this study was to develop a high-throughput RP-UHPSFC/MS method for the determination of nonpolar lipid classes, namely, fatty acids (FA), sterols (ST), sterol esters, mono- (MG), di- (DG), and tri-acylglycerols in plasma and serum samples. The RP-UHPSFC method using connection of two sub‑2‑mm particles columns in series (150 + 100 mm × 3.0 mm; 1.8 mm) was coupled to high-resolution mass spectrometry. The method was optimized based on 86 standards, resulting in high separation efficiency, and total run time was 18 min. Moreover, the method enables the separation of positional and cis/trans isomers. Several extraction protocols were investigated, where the use of selective extraction of nonpolar	supercritical fluid chromatography; lipidomics; nonpolar lipids; mass spectrometry