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Chemical derivatization of lipid functional groups for analysis of human plasma by RP-UHPLC/MS

Authors: Peterka Ondřej | Jirásko Robert | Bureš Filip | Holčapek Michal

Year: 2020

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	Chemical derivatization of lipid functional groups for analysis of human plasma by RP-UHPLC/MS	The chemical derivatization of lipid functional group can improve extraction efficiency, chromatographic separation, and sensitivity. Acetyl chloride is one of the most frequently used derivatization agent due to high reactivity, stability of products, and wide application range for various functional groups. A strongly alkaline and non-aqueous medium is necessary for the lipid derivatization, because the derivatizing agent may undergo hydrolysis, therefore pyridine was chosen as the base. Reversed phase ultrahigh-performance liquid chromatography (RP-UHPLC) coupled to mass spectrometry method separated lipids according to the equivalent carbon number and tandem mass spectrometry in positive and negative ion modes was used for the identification of lipid species. The behavior of lipid classes was observed on individual internal standard (IS), whereby primary amine, hydroxyl, and carboxylic groups produce amides and esters. Reaction condition, such as derivatization agent/base molar ratio, reaction time and temperature, were optimized on the spiked deproteinized human plasma. Liquid-liquid extraction has been included to avoid the contamination of the mass spectrometer by excessive amount of agent and base. The mixture of 37 lipid species representing 21 lipid classes was tested and the method was successful applied for 16 lipid classes providing expected derivatization products. The optimized derivatization method was applied for human plasma and the derivatization significantly improved the extraction efficiency and sensitivity for glycerolipids, sphingolipids, and most of phospholipids.	Lipidomics, Chemical Derivatization, RP-UHPLC, Mass Spectrometry