Authors: Michal Holčapek Magdaléna Ovčačíková Miroslav Lísa Eva Cífková Tomáš Hájek
Publish Date: 2015/02/19
Volume: 407, Issue: 17, Pages: 5033-5043
Abstract
A new continuous comprehensive twodimensional liquid chromatography–electrospray ionization mass spectrometry method has been developed for the lipidomic characterization of complex biological samples The reversedphase ultrahighperformance liquid chromatography with a C18 column 150 mm × 1 mm 17 μm used in the first dimension makes the separation of numerous lipid species differing in their hydrophobic part of the molecule mainly fatty acyl chain lengths and the number and positions of double bonds possible Coeluted lipid species in the first dimension are resolved by the fast hydrophilic interaction liquid chromatography separation 50 mm × 3 mm 27 μm core–shell particles of lipid classes according to their different polarities in the second dimension Retention times in both dimensions accurate m/z values and tandem mass spectra provide high confidence in the identification of lipid species The retention behavior of individual lipids in reversedphase mode follows the equivalent carbon number pattern which provides an additional tool for unambiguous identification This analytical method is applied for the lipidomic characterization of total lipid extracts of human plasma and porcine brain samples which resulted in the identification of 143 lipid species from four lipid categories and ten lipid classesThis work was supported by ERC CZ project no LL1302 sponsored by the Ministry of Education Youth and Sports of the Czech Republic EC acknowledges the support of project no CZ107/2300/300021 sponsored by the Ministry of Education Youth and Sports of the Czech Republic
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