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Two-dimensional ultra high-performance liquid chromatography – mass spectrometry characterization of comprehensive samples of human plasma
Authors: Ovčačíková Magdaléna | Cífková Eva | Lísa Miroslav | Holčapek Michal
Year: 2014
Type of publication: ostatní do riv
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
cze Two-dimensional ultra high-performance liquid chromatography – mass spectrometry characterization of comprehensive samples of human plasma Lipids belong among one of the most important constituents forming all biological tissues and body fluids. Their interaction with surrounding cells has a crucial role in living organisms. The changes of lipid metabolism play a significant role in many serious human diseases, such as cancer, atherosclerosis, obesity, diabetes mellitus, cardiovascular disease and Alzheimer's disease. Lipidomics is a subset of metabolomics, which contributes to the understanding how lipids work in a biological system (cellular physiology and pathology) [1-3]. One-dimensional chromatographic separation may be not sufficient for the detailed lipidomic characterization due to various types of isomerism. Therefore two-dimensional ultra high-performance liquid chromatography - mass spectrometry (2D-UHPLC/MS) can be used to increase the number of compounds separated and quantitated in a single run [3, 4]. Our work is focused on the development of a new comprehensive 2D-UHPLC method with electrospray ionization mass spectrometry (ESI-MS) for the lipidomic characterization of complex biological samples. As a first dimension, reversed-phase liquid chromatography (RP-LC) with a C18 column is used, where lipids are separated according to the acyl chain length and the number of double bonds. Hydrophilic interaction liquid chromatography (HILIC) mode using a silica column is used as the second dimension. In HILIC separation mode, lipids are separated into individual lipid classes according to their polarity and electrostatic interactions. Fractions from the first dimension column are transferred to the second dimension column using a two-position eight-port switching valve, operated under on-line conditions in combination with MS detection. Acknowledgement This work was supported by ERC CZ grant project LL1302 (Ministry of Education, Youth and Sports of the Czech Republic).
eng Two-dimensional ultra high-performance liquid chromatography – mass spectrometry characterization of comprehensive samples of human plasma Lipids belong among one of the most important constituents forming all biological tissues and body fluids. Their interaction with surrounding cells has a crucial role in living organisms. The changes of lipid metabolism play a significant role in many serious human diseases, such as cancer, atherosclerosis, obesity, diabetes mellitus, cardiovascular disease and Alzheimer's disease. Lipidomics is a subset of metabolomics, which contributes to the understanding how lipids work in a biological system (cellular physiology and pathology) [1-3]. One-dimensional chromatographic separation may be not sufficient for the detailed lipidomic characterization due to various types of isomerism. Therefore two-dimensional ultra high-performance liquid chromatography - mass spectrometry (2D-UHPLC/MS) can be used to increase the number of compounds separated and quantitated in a single run [3, 4]. Our work is focused on the development of a new comprehensive 2D-UHPLC method with electrospray ionization mass spectrometry (ESI-MS) for the lipidomic characterization of complex biological samples. As a first dimension, reversed-phase liquid chromatography (RP-LC) with a C18 column is used, where lipids are separated according to the acyl chain length and the number of double bonds. Hydrophilic interaction liquid chromatography (HILIC) mode using a silica column is used as the second dimension. In HILIC separation mode, lipids are separated into individual lipid classes according to their polarity and electrostatic interactions. Fractions from the first dimension column are transferred to the second dimension column using a two-position eight-port switching valve, operated under on-line conditions in combination with MS detection. Acknowledgement This work was supported by ERC CZ grant project LL1302 (Ministry of Education, Youth and Sports of the Czech Republic). liquid chromatography; mass spectrometry; human plasma

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