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Quantitative Analysis and Lipidomic Profiling of Human Serum of Pancreatic Cancer Patients Focused on Less Abundant Lipid Classes by HILIC-UHPLC/MS
Authors: Peterka Ondřej | Maccelli Alessandro | Jirásko Robert | Vaňková Zuzana | Idkowiak Jakub | Wolrab Denise | Holčapek Michal | Hrstka Roman
Year: 2023
Type of publication: ostatní - přednáška nebo poster
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
eng Quantitative Analysis and Lipidomic Profiling of Human Serum of Pancreatic Cancer Patients Focused on Less Abundant Lipid Classes by HILIC-UHPLC/MS Lipidomics is a field that focusses on the study of lipids in living systems. Lipids play an important role in all organisms, such as membrane components, energy storage, precursors for metabolic processes, and signaling, which predestines lipids for biomarkers of cancer, cardiovascular diseases, and neurodegenerative diseases. The comprehensive analysis of high- and low-abundant lipid classes together is extremely challenging because injection of concentrated extract leads to the saturation of the detector, ion suppression, or contamination of the mass spectrometer, while signals for low-abundant lipids are missing in diluted extracts. Moreover, the selective extraction is almost impossible because of the structural similarities. However, the analysis of all lipid classes within their biosynthetic pathways is essential for understanding lipid dysregulations in human metabolism caused by various diseases, such as cancer. Hydrophilic interaction liquid chromatography separates lipids according to the polar head group, which allows the switch of highly abundant lipid classes to waste, which enables the injection of concentrated lipidomic extract. An Agilent 1290 Infinity series LC coupled to a Xevo G2-XS QTOF mass spectrometer was used and the new separation method using the Type-C Cogent Silica column (150×2.1 mm; 2.1 µm) column with the total run time of 20 min was developed. Deproteinization using a mixture of BuOH/MeOH was used for the sample preparation. The method is based on the switch of inorganic salts in the void volume and high-abundance lipid classes (cholesterol esters, triacylglycerols, and phosphatidylcholines) to the waste, preventing contamination of the mass spectrometer. In total, 246 lipid species from 24 lipid subclasses using both positive and negative ion modes were identified in pooled human plasma. The method was fully validated and almost two hundred lipids were quantified using an internal standard for each lipid class. Our laboratory-made

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