Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

Zaměstnanci
|
Studenti
English

Publikace detail

UHPSFC/MS Lipidomic Analysis of biological samples

Autoři: Wolrab Denise | Peterka Ondřej | Hrstka Roman | Holčapek Michal

Rok: 2018

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	UHPSFC/MS Lipidomic Analysis of biological samples	Lipids are biologically active compounds with several biological functions. They are involved in energy storage, function as signaling molecules and present constituents of cell membranes. The analysis and quantification of lipids in biological samples like human plasma or serum may allow the differentiation of healthy and diseased donors. However, the chromatographic analysis is challenging, due to the structural and chemical diversity of lipids in biological samples. Supercritical fluid chromatography allows a fast separation of non-polar and polar lipid classes and consequently also the quantification by employing an internal standard for each lipid class. The assessment of small differences in the lipid profile in human body fluids requires robust and reproducible methodology. Therefore, a partial validation for the analysis of various lipid classes, like cholesteryl esters, triacylglycerols, diacylglycerols, monoacylglycerols, ceramides, phosphatidylcholines, lysophosphatidylcholines, and sphingomyelins from human plasma and serum using ultrahigh-performance supercritical fluid chromatography (UHPSFC/MS) was performed. Strategies for the evaluation of the method performance were developed, and these results were further applied for lipid analysis of human body fluids of various origin. Liquid-liquid extraction was employed for sample preparation. UHPSFC/MS measurements were carried out on an Acquity Ultra Performance Convergence Chromatography (UPC2) System coupled to a hybrid quadrupole traveling wave ion mobility time of flight mass spectrometer Synapt G2 Si from Waters. For validation purposes, representative samples of different origin, like gender, age and health state were mixed and used for the determination of the following parameters: selectivity, accuracy, repeatability, intermediate precision, extraction recovery, process efficiency, and matrix effect. Aliquots of the pooled sample were spiked before and after extraction at different concentrations w	UHPSFC/MS; Lipidomic; Serum