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Quantitative Lipidomic Analysis of Biological Samples: Optimization and Application for the Analysis of Clinical Samples using UHPSFC/MS
Authors: Wolrab Denise | Chocholoušková Michaela | Peterka Ondřej | Jirásko Robert | Hrstka Roman | Vrána David | Melichar Bohuslav | Holčapek Michal
Year: 2019
Type of publication: ostatní - přednáška nebo poster
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
eng Quantitative Lipidomic Analysis of Biological Samples: Optimization and Application for the Analysis of Clinical Samples using UHPSFC/MS The development of a sensitive and specific quantitative lipidomic analysis method for the differentiation of healthy and cancerous samples is aimed. Special focus was put on the reliable quantitation of over 100 lipid species using lipidomic profiling. Consequently, the used internal standard mixture regarding structure and concentrations and the sample preparation protocol were optimized. Several liquid-liquid extraction protocols, the influence of various solvents for dissolving the extracts, the influence of the pH during extraction, the influence of the sample dilution solvent were evaluated employing hydrophilic interaction liquid chromatography (HILIC-UHPLC/MS) and ultrahigh-performance supercritical fluid chromatography hyphenated to mass spectrometry (UHPSFC/MS). The differences in the validation parameters, such as matrix effect, recovery rate, precision, accuracy, repeatability and selectivity obtained by UHPLC/MS and UHPSFC/MS were investigated. The optimized sample preparation protocol was applied for the analysis of clinical plasma and serum samples and the SRM 1950 plasma. The concentrations for the different cohorts obtained with both methods were compared to each other and literature [1, 2]. The lipidomic profiles depending on the used blood collection tubes such as Lithium heparin plasma tubes, K2 EDTA plasma tubes and Z serum clot activator tubes were evaluated using both methods. Liquid-liquid extraction using a slightly modified Folch procedure, and basic conditions were found superior for sample preparation with regard to extraction recovery of various lipid classes. The extract dissolving solvent of choice was a CHCl3/MeOH (1/1 v/v) mixture. Results show that UHPLC/MS and UHPSFC/MS are comparable, robust, precise and fast methods for the quantitative analysis of lipid species, whereby UHPLC/MS is more suited for the analysis of polar analytes and UHPSFC/MS for non-polar and polar lipid species.

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