Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

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Lipidomic Profiling of Low Abundant Lipids in Human Blood by HILIC/MS

Autoři: Peterka Ondřej | Jirásko Robert | Vaňková Zuzana | Maccelli Alessandro | Idkowiak Jakub | Holčapek Michal | Hrstka Roman

Rok: 2023

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	Lipidomic Profiling of Low Abundant Lipids in Human Blood by HILIC/MS	The full characterization of the human lipidome is an actual challenge in the lipidomics, but lipids are highly diverse biomolecules with various concentration levels in the biological matrix, making it difficult to determine large numbers of lipids in one analytical method. Injection of the concentrated extract leads to the saturation of the detector or contamination of the mass spectrometer, and signal of a low-abundant lipids is missing in diluted extracts. However, the lipidomic composition usually reflects the actual state of the organism caused by various diseases, such as cancer, neurodegenerative diseases, and cardiovascular diseases. Analysis of all lipid classes within their biosynthetic pathways is therefore essential for understanding lipid dysregulations in human metabolism. The new hydrophilic interaction liquid chromatography – mass spectrometry (HILIC/MS) method was developed with a focus on the less abundant lipid species in human plasma and serum. The chromatography and mass spectrometry conditions were optimized using 32 standards representing the same number of lipid subclasses. The method is based on the switch of highly abundant lipid classes to the waste, preventing contamination of the mass spectrometer, which enables injection of concentrated lipidomic extract. Highly confident identification based on the combination of high mass accuracy, retention time and retention dependencies in positive and negative ion modes was used, resulting in the identification of 246 lipid species from 24 lipid subclasses in pooled human plasma. The method was validated based on human plasma spiked by 31 internal standards and used for quantitative analysis of SRM 1950 human plasma. In total, 169 lipid species were quantified, and generated results of obtained concentrations were comparable with literature. Finally, the method was used for the investigation of the lipidomic profile of serum samples obtained from 25 pancreatic cancer patients (PDAC) and 25 hea