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Lipidomic Analysis of Biological Samples: Comparison of Liquid Chromatography, Supercritical Fluid Chromatography and Direct Infusion Mass Spectrometry Methods
Autoři: Lísa Miroslav | Cífková Eva | Khalikova Maria | Ovčačíková Magdaléna | Holčapek Michal
Rok: 2017
Druh publikace: ostatní - přednáška nebo poster
Strana od-do: nestránkováno
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Jazyk Název Abstrakt Klíčová slova
eng Lipidomic Analysis of Biological Samples: Comparison of Liquid Chromatography, Supercritical Fluid Chromatography and Direct Infusion Mass Spectrometry Methods Lipidomic analysis of biological samples in a clinical research represents challenging task for analytical methods given by the large number of samples and their extreme complexity. In this work, we compare three analytical methods in terms of comprehensiveness, sample throughput, and validation results for the lipidomic analysis of biological samples represented by tumor tissue, surrounding normal tissue, plasma, and erythrocytes of kidney cancer patients. Methods are compared in one laboratory using the identical analytical protocol and instrumentation to ensure comparable conditions. Ultrahigh-performance liquid chromatography/mass spectrometry (UHPLC/MS) method in hydrophilic interaction liquid chromatography mode and direct infusion MS (DI-MS) method are used for this comparison as the most widely used methods for the lipidomic analysis together with ultrahigh-performance supercritical fluid chromatography/MS (UHPSFC/MS) method showing promising results in metabolomics analyses. The nontargeted analysis of pooled samples is performed using all tested methods and 610 lipid species within 23 lipid classes are identified. DI method provides the most comprehensive results due to identification of some polar lipid classes, which are not identified by UHPLC and UHPSFC methods. On the other hand, UHPSFC provides an excellent sensitivity for less polar lipid classes and the highest sample throughput within 10 min method time. The sample consumption of DI method is 125 times higher than for other methods, while only 40 μL of organic solvent is used for one sample analysis compared to 3.5 mL and 4.9 mL in case of UHPLC and UHPSFC methods, respectively. Methods are validated for the quantitative lipidomic analysis of plasma samples with one internal standard for each lipid class. Results show applicability of tested methods for the lipidomic analysis of biological samples depending on the analysis requirements. Lipidomic analysis; Supercritical Fluid Chromatography; Direct Infusion; Mass Spectrometry

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