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Lipidomic Analysis of Human Serum of Pancreatic Cancer Patients with Focus 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	Lipidomic Analysis of Human Serum of Pancreatic Cancer Patients with Focus on Less Abundant Lipid Classes by HILIC-UHPLC/MS	Introduction (Limit 120 words) Hydrophilic interaction liquid chromatography separates lipids according to the polar head group, which leads to coelution of lipids of all lipid species within one lipid class, including the exogenous IS. This approach guarantees the same matrix effect and ionization efficiency necessary for accurate quantitation. However, LC/MS analyses of lipids are often challenging, depending on the various abundances of lipids within a biological sample. Injection of concentrated extract leads to saturation of the detector or contamination of the mass spectrometer, and signal of low-abundant lipids is missing in diluted extracts. 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. Methods (Limit 120 words) Human serum (25 μL) was deproteinized using 250 μL of BuOH/MeOH mixture placed in an ultrasonic bath for 15 min and purified using a 0.25 μm cellulose filter. We used an Agilent 1290 Infinity series LC coupled to a Xevo G2-XS QTOF mass spectrometer. Type-C Cogent Silica column (150×2.1 mm; 2.1 µm) column, flow rate 0.4 mL/min, injection 1 μL, column temperature 40 ° C was employed. The gradient elution was used, where the mobile phase A is acetonitrile and B 40 mmol/L of aqueous ammonium formate (pH 4.65). A valve was switched to waste during intervals 0–1.35 min and 11.40–12.20 min due to the elimination of high-abundant lipid classes. The total run time is 20 min. Preliminary data (Limit 300 words) The goal of our work is the analysis of wide-range lipid classes combined with simple sample preparation. The method is based on the switch of inorganic salts in the void volume and high-abundance lipid classes (CE, TG, DG, and PC) to the waste, preventing contamination of the mass spectrometer, which enables injection of concentrated lipidomic extract. Moreover, the method does not require sophisticate