UHPSFC/ESI-MS Analysis of Lipids

  • Miroslav Lísa
  • Michal HolčapekEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1730)


This new analytical approach for high-throughput and comprehensive lipidomic analysis of biological samples using ultrahigh-performance supercritical fluid chromatography (UHPSFC) with electrospray ionization–mass spectrometry (ESI-MS) is based on lipid class separation using 1.7 μm particle bridged ethylene hybrid silica columns and a gradient of methanol–water–ammonium acetate mixture as a modifier. The method enables a fast separation of 30 nonpolar and polar lipid classes within 6-min analysis time covering six main lipid categories including fatty acyls, glycerolipids, glycerophospholipids, sphingolipids, sterols, and prenols. Individual lipid species within lipid classes are identified based on positive- and negative-ion full scan and tandem mass spectra measured with high mass accuracy and high resolving power. The method is used for the quantitative analysis of lipid species in biological tissues using internal standards for each lipid class. This high-throughput, comprehensive, and accurate UHPSFC/ESI-MS method is suitable for the lipidomic analysis of large sample sets in clinical research.

Key words

Supercritical fluid chromatography Ultrahigh-performance supercritical fluid chromatography Mass spectrometry Electrospray ionization Lipids Lipidomics Glycerolipids Glycerophospholipids Sphingolipids 



This work was supported by the grant project LL1302 sponsored by the Ministry of Education, Youth and Sports of the Czech Republic.


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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Department of Analytical Chemistry, Faculty of Chemical TechnologyUniversity of PardubicePardubiceCzech Republic

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