University of Pardubice Faculty of Chemical Technology

University

Employees
|
Students
Česky

Publication detail

Qualitative and Quantitative Lipidomic Analysis of Human Plasma Using the RP-UHPLC/MS Approach

Authors: Vaňková Zuzana | Peterka Ondřej | Idkowiak Jakub | Jirásko Robert | Holčapek Michal

Year: 2022

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	Qualitative and Quantitative Lipidomic Analysis of Human Plasma Using the RP-UHPLC/MS Approach	Lipidomic analysis is one of the constantly evolving omics methods that examines multiple biological functions of lipids. Lipid dysregulation is associated with serious diseases such as diabetes mellitus, cardiovascular disease, cancer, etc. Their analysis is mediated by the combination of mass spectrometry (MS) with a direct infusion or prior separation. One of the most widely used approaches is the reversed-phase ultrahigh-performance liquid chromatography (RP-UHPLC) method, which separates lipid species according to the polarity, length of the fatty acyl/alkyl chain, and number and position of double bonds. Our goal was to develop the RP-UHPLC/MS method for the quantitative analysis of a large number of lipid species from multiple polar and nonpolar lipid classes with highly confident identification. For the identification of lipids, the following criteria were set: a) a high mass accuracy of m/z values, where full scan and tandem mass spectra were measured using a high-resolution QTOF mass analyzer in both polarity modes of ESI, b) ability to find characteristic ions in MS and fragment ions in MS/MS, and c) regularity in chromatographic retention dependences for individual lipid species. In addition, the retention behavior of individual homologous lipid series was verified using graphs of dependence of the retention times on the carbon number or the number of double bond(s) in fatty acyl chains. A low-resolution Q-Trap instrument was used for the validation and quantitative analysis of identified lipid species in human plasma sample, using appropriate internal standards. For individual lipid species, the characteristic and unique transitions of molecular masses and fragment ions were set and measured with the multiple reaction monitoring (MRM) approach. Specific response factors have been used to calculate more exact concentration of nonpolar lipid classes (TG, DG, MG, and CE). Our optimized and validated RP-UHPLC/MS method is a unique tool for the analysis of t	Lipidomic; Plasma; RP-UHPLC/MS