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Analytical and Bioanalytical Chemistry

, Volume 407, Issue 17, pp 5033–5043 | Cite as

Continuous comprehensive two-dimensional liquid chromatography–electrospray ionization mass spectrometry of complex lipidomic samples

  • Michal HolčapekEmail author
  • Magdaléna Ovčačíková
  • Miroslav Lísa
  • Eva Cífková
  • Tomáš Hájek
Paper in Forefront
Part of the following topical collections:
  1. Lipidomics

Abstract

A new continuous comprehensive two-dimensional liquid chromatography–electrospray ionization mass spectrometry method has been developed for the lipidomic characterization of complex biological samples. The reversed-phase ultra-high-performance liquid chromatography with a C18 column (150 mm × 1 mm, 1.7 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, 2.7 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 reversed-phase 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 classes.

Graphical Abstract

2D-LC/ESI-MS of porcine brain lipid extract

Keywords

Lipids Lipidomics Comprehensive 2D liquid chromatography Mass spectrometry Plasma Brain 

Notes

Acknowledgments

This work was supported by ERC CZ project no. LL1302 sponsored by the Ministry of Education, Youth, and Sports of the Czech Republic. E.C. acknowledges the support of project no. CZ.1.07/2.3.00/30.0021 sponsored by the Ministry of Education, Youth, and Sports of the Czech Republic.

Supplementary material

216_2015_8528_MOESM1_ESM.pdf (461 kb)
ESM 1 (PDF 461 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Michal Holčapek
    • 1
    Email author
  • Magdaléna Ovčačíková
    • 1
  • Miroslav Lísa
    • 1
  • Eva Cífková
    • 1
  • Tomáš Hájek
    • 1
  1. 1.Department of Analytical Chemistry, Faculty of Chemical TechnologyUniversity of PardubicePardubiceCzech Republic

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