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UHPSFC/MS: A powerful Tool for Lipidomic Quantitation in Clinical Samples

Authors: Wolrab Denise | Peterka Ondřej | Jirásko Robert | Holčapek Michal

Year: 2018

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	UHPSFC/MS: A powerful Tool for Lipidomic Quantitation in Clinical Samples	Lipids are biologically active compounds with several biological functions. They are involved in energy storage, function as signaling molecules and present constituents of cell membranes. A dysregulation is often related to serious diseases, e.g., various types of cancer. The analysis and quantification of lipids in biological samples, like human plasma or serum, may allow the differentiation of healthy and diseased donors. However, the chromatographic analysis is challenging due to the structural and chemical diversity as well as concentration differences of lipids in biological samples. Ultrahigh-performance supercritical fluid chromatography (UHPSFC) allows a fast separation of nonpolar and polar lipid classes and consequently also the quantitation by employing an internal standard for each lipid class and mass spectrometry (MS) detection. Liquid-liquid extraction was employed for the extraction of lipids from human serum. UHPSFC/MS measurements were carried out on an Acquity Ultra Performance Convergence Chromatography (UPC2) system coupled to a hybrid quadrupole traveling wave ion mobility time of flight mass spectrometer Synapt G2 Si from Waters (Milford, USA). Lipid classes were separated according to their polar head groups employing an Acquity BEH UPC2 column in gradient mode. Positive ion ESI mode in the resolution mode was employed for analysis of lipid species belonging to glycerophospholipids, sphingolipids, and glycerolipids. For validation purpose, we determined LOD, LOQ, matrix effect, recovery, repeatability as well as reproducibility for 8 lipid subclasses using appropriate exogenous internal standards for each lipid subclass. Results show slight differences in the method performance parameters depending on the nature of the head group of the lipid subclass. Furthermore, we developed a workflow for monitoring, method performance and sample preparation quality among the study in order to ensure reliable data quality. Therefore, a QC sample, which i	UHPSFC/MS; Lipidomic; Serum; Cancer