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A direct and simple method of coupling matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) to thin-layer chromatography (TLC) for the analysis of phospholipids from egg yolk

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Abstract

Although the most important application of matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) is “proteomics,” there is growing evidence that this soft ionization method is also useful for phospholipid (PL) analysis. Although all PLs are detectable by MALDI-TOF MS, some lipid classes, particularly those with quaternary amines such as phosphatidylcholines (PCs), are more sensitively detected than others, and these suppress the signals of less sensitively detected PLs when complex mixtures are analyzed. Therefore, a separation of the total organic extract into individual lipid classes is necessary. As MALDI uses a solid sample, the direct evaluation of thin-layer chromatography (TLC) plates is possible. We report here on a method of directly coupling MALDI-TOF MS and TLC that can be easily implemented on commercially available MALDI-TOF devices. A total extract of hen egg yolk is used as a simple PL mixture to demonstrate the capabilities of this method. It will be shown that “clean” spectra without any major contributions from fragmentation products and matrix peaks can be obtained, and that this approach is even sensitive enough to detect the presence of PLs at levels of less than 1% of the total extract.

TLC/MALDI coupling is performed in the following way: Phospholipids are separated by TLC in the conventional way and stained with the dye PRIMULINE that binds non-covalently to the acyl residues of phospholipids. Afterwards, the spots of interest are soaked with matrix and the TLC plate mounted onto a special adapter target. This target is introduced into the mass spectrometer and analyzed in the conventional manner

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Acknowledgements

This work was supported by the German Research Council (DFG Schi 476/5-1) and the Federal Ministry of Education and Research (Grant BMBF 0313836).

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Authors and Affiliations

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

    Beate Fuchs, Jürgen Schiller & Rosmarie Süß

  2. Bruker Daltonik GmbH, Fahrenheitstrasse 4, 28359, Bremen, Germany

    Martin Schürenberg & Detlev Suckau

Authors
  1. Beate Fuchs
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  2. Jürgen Schiller
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  3. Rosmarie Süß
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  4. Martin Schürenberg
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  5. Detlev Suckau
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Corresponding author

Correspondence to Jürgen Schiller.

Additional information

Beate Fuchs, Jürgen Schiller and Rosmarie Süß contributed equally to this work.

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Fuchs, B., Schiller, J., Süß, R. et al. A direct and simple method of coupling matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) to thin-layer chromatography (TLC) for the analysis of phospholipids from egg yolk. Anal Bioanal Chem 389, 827–834 (2007). https://doi.org/10.1007/s00216-007-1488-4

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  • DOI: https://doi.org/10.1007/s00216-007-1488-4

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