Skip to main content

UHPSFC/ESI-MS Analysis of Lipids

  • Protocol
Clinical Metabolomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1730))

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 79.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 99.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 129.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Nováková L, Perrenoud AG-G, Francois I, West C, Lesellier E, Guillarme D (2014) Modern analytical supercritical fluid chromatography using columns packed with sub-2 micrometer particles: a tutorial. Anal Chim Acta 824:18–35. https://doi.org/10.1016/j.aca.2014.03.034

    Article  PubMed  CAS  Google Scholar 

  2. Čajka T, Fiehn O (2014) Comprehensive analysis of lipids in biological systems by liquid chromatography-mass spectrometry. Trends Anal Chem 61:192–206. https://doi.org/10.1016/j.trac.2014.04.017

    Article  CAS  Google Scholar 

  3. Lesellier E, Gaudin K, Chaminade P, Tchapla A, Baillet A (2003) Isolation of ceramide fractions from skin sample by subcritical chromatography with packed silica and evaporative light scattering detection. J Chromatogr A 1016(1):111–121. https://doi.org/10.1016/s0021-9673(03)01323-2

    Article  PubMed  CAS  Google Scholar 

  4. Lísa M, Holčapek M (2015) High-throughput and comprehensive lipidomic analysis using ultrahigh-performance supercritical fluid chromatography-mass spectrometry. Anal Chem 87(14):7187–7195. https://doi.org/10.1021/acs.analchem.5b01054

    Article  PubMed  CAS  Google Scholar 

  5. Schuhmann K, Almeida R, Baumert M, Herzog R, Bornstein SR, Shevchenko A (2012) Shotgun lipidomics on a LTQ Orbitrap mass spectrometer by successive switching between acquisition polarity modes. J Mass Spectrom 47(1):96–104. https://doi.org/10.1002/jms.2031

    Article  PubMed  CAS  Google Scholar 

  6. Wiesner P, Leidl K, Boettcher A, Schmitz G, Liebisch G (2009) Lipid profiling of FPLC-separated lipoprotein fractions by electrospray ionization tandem mass spectrometry. J Lipid Res 50(3):574–585. https://doi.org/10.1194/jlr.D800028-JLR200

    Article  PubMed  CAS  Google Scholar 

  7. Fairchild JN, Brousmiche DW, Hill JF, Morris MF, Boissel CA, Wyndham KD (2015) Chromatographic evidence of Silyl ether formation (SEF) in supercritical fluid chromatography. Anal Chem. https://doi.org/10.1021/ac5035709

  8. Lísa M, Cífková E, Holčapek M (2011) Lipidomic profiling of biological tissues using off-line two-dimensional high-performance liquid chromatography mass spectrometry. J Chromatogr A 1218(31):5146–5156. https://doi.org/10.1016/j.chroma.2011.05.081

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

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

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Michal Holčapek .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Lísa, M., Holčapek, M. (2018). UHPSFC/ESI-MS Analysis of Lipids. In: Giera, M. (eds) Clinical Metabolomics. Methods in Molecular Biology, vol 1730. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7592-1_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7592-1_5

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7591-4

  • Online ISBN: 978-1-4939-7592-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics