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Thin-layer chromatography combined with MALDI-TOF-MS and 31P-NMR to study possible selective bindings of phospholipids to silica gel

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Abstract

High-performance thin-layer chromatography (HPTLC) is a highly established separation method in the field of lipid and (particularly) phospholipid (PL) research. HPTLC is not only used to identify certain lipids in a mixture but also to isolate lipids (preparative TLC). To do this, the lipids are separated and subsequently re-eluted from the silica gel. Unfortunately, it is not yet known whether all PLs are eluted to the same extent or whether some lipids bind selectively to the silica gel. It is also not known whether differences in the fatty acyl compositions affect the affinities to the stationary phase. We have tried to clarify these questions by using a readily available extract from hen egg yolk as a selected example of a lipid mixture. After separation, the complete lanes or selected spots were eluted from the silica gel and investigated by a combination of MALDI-TOF MS and 31P NMR spectroscopy. The data obtained were compared with the composition of the total extract (without HPTLC). Although there were significant, solvent-dependent losses in the amount of each lipid, the relative composition of the mixture remained constant; there were also only very slight changes in the fatty acyl compositions of the individual PL classes. Therefore, lipid isolation by TLC may be used without any risk of major sample alterations.

Phospholipids separated by HPTLC were subsequently re-eluted from the plate and characterized by different methods. Spectra in blue represent the original mixture that was applied onto the HPTLC plate, while the red spectra represent the phospholipids subsequent to HPTLC. Although there were significant losses in the amount of each phospholipid, no changes in the relative and fatty acyl compositions occurred.

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Acknowledgements

This work was supported by the German Research Council (TR 67, project A2 and SCHI 476/12-1). Additionally, this publication was supported by LIFE—Leipzig Research Center for Civilization Diseases, University of Leipzig. LIFE is funded by the European Regional Development Fund (ERFD) and by means of the Free State of Saxony within the framework of the Excellence Initiative.

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Authors and Affiliations

  1. LIFE- Leipzig Research Center for Civilization Diseases, University of Leipzig, 04103, Leipzig, Germany

    Kristin Teuber & Jürgen Schiller

  2. Faculty of Medicine, Institute of Medical Physics and Biophysics, University of Leipzig, Härtelstrasse 16-18, 04107, Leipzig, Germany

    Kristin Teuber, Thomas Riemer & Jürgen Schiller

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  1. Kristin Teuber
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  2. Thomas Riemer
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  3. Jürgen Schiller
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Correspondence to Kristin Teuber.

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Published in the special issue Mass Spectrometry (DGMS 2010) with Guest Editors Andrea Sinz and Jürgen Schmidt.

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Teuber, K., Riemer, T. & Schiller, J. Thin-layer chromatography combined with MALDI-TOF-MS and 31P-NMR to study possible selective bindings of phospholipids to silica gel. Anal Bioanal Chem 398, 2833–2842 (2010). https://doi.org/10.1007/s00216-010-4064-2

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  • DOI: https://doi.org/10.1007/s00216-010-4064-2

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