University of Pardubice Faculty of Chemical Technology

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Novel derivatization method for the determination of sterols, prenols, and neutral lipids by RP-UHPLC/MS/MS in negative ion mode

Authors: Kadyrbekova Yasmin | Peterka Ondřej | Jirásko Robert | Vaňková Zuzana | Holčapek Michal

Year: 2023

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	Novel derivatization method for the determination of sterols, prenols, and neutral lipids by RP-UHPLC/MS/MS in negative ion mode	Lipidomics studies the structure, function, and metabolism of lipids. Lipids are essential components of all living organisms and play critical roles in cellular processes such as energy storage, signaling, and membrane structure. Cholesterol plays a crucial role in animal cells by regulating membrane fluidity as well as the synthesis of hormones, such as estrogen, testosterone, and cortisol. Prenols are derived from the mevalonic acid pathway, which is responsible for the biosynthesis of a wide range of biomolecules, including coenzymes and vitamins. Dysregulation of lipid synthesis has an impact on health, therefore, the study of lipids like sterols and prenols can lead to a better understanding of the role of these lipids in the pathogenesis of diseases. In lipidomics, mass spectrometry (MS) coupled with chromatography allows for high-resolution separation and sensitive detection. The combination of reversed-phase (RP) chromatography and selected reaction monitoring (SRM) transitions on QqQ instrument provides the separation of isomers and high sensitivity. Some lipid classes are not appropriate for this approach because they do not produce selective fragments. However, such a problem can be solved by the derivatization approach. The novel derivatization method using 3-(chlorosulfonyl)benzoic acid, which had never been used before for the derivatization of lipids, was developed for the determination of sterols, prenols, and neutral lipids. The reaction enables the detection of these lipid classes in the negative - ion mode, which minimizes the in-source fragmentation common in the positive - ion mode, leading to higher sensitivity. The RP-UHPLC/MS/MS method was optimized based on 30 derivatized standards from 4 lipid classes (sterols, prenols, monoacylglycerols, and diacylglycerols). The derivatization reaction was thoroughly optimized using spiked pooled plasma (9 internal standard from 4 lipid classes), and the molar ratio 6:1 of pyridine with derivatization