Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

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Studenti
English

Publikace detail

Quantitation of lipids using HILIC-UHPLC/MS: An optimized and validated method for the analysis of biological samples.

Autoři: Chocholoušková Michaela | Wolrab Denise | Jirásko Robert | Vrána David | Melichar Bohuslav | Holčapek Michal

Rok: 2019

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	Quantitation of lipids using HILIC-UHPLC/MS: An optimized and validated method for the analysis of biological samples.	The aim of this project is the development of a robust and accurate method for the determination of lipid profiles in human body fluids and the application of those for the analysis of blood samples of healthy volunteers and patients suffering from kidney cancer. A special focus is given to the optimization of the sample preparation protocol for the analysis of lipids in biological samples, the choice of internal standards (structure and concentration), the extraction solvents for liquid-liquid extraction (Folch extraction, methyl-tert-butyl ether extraction, butanol/methanol extraction, etc.), the influence of pH during the extraction (ammonium carbonate, ammonium acetate, etc.), and the influence of different solvent mixtures for dissolving the extracts are investigated. All experiments are performed using hydrophilic interaction liquid chromatography coupled to high-resolution mass spectrometry (HILIC-UHPLC/MS), and the optimized protocol is fully validated. The best results with regard to the recovery are obtained by employing a slightly modified double Folch extraction using basic conditions and the reconstitution of the extracts in a mixture of CHCl3/MeOH (1:1). The lipid concentration results of the optimized method applied for the analysis of the SRM 1950 reference plasma are in agreement with established values from the literature [1, 2]. All experiments are also performed using ultra-high performance supercritical fluid chromatography coupled to high-resolution mass spectrometry (UHPSFC/MS), and results are compared to each other using validation parameters and statistical analysis.