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Fast counter-electroosmotic capillary electrophoresis–time-of-flight mass spectrometry of hyaluronan oligosaccharides

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Abstract

Fast capillary electrophoresis–mass spectrometry measurements under counter-electroosmotic analyte migration conditions are presented. Efficient separations of a homologous series of six hyaluronan oligosaccharides (comprising 1–6 hyalobiuronic acid moieties) could be completed in 65 s. Separations were achieved in short-length fused silica capillaries under high electric field strengths of up to 1.25 kV·cm−1. Capillary inner diameters ranging from 5 to 50 μm were investigated, resulting in an optimal value of 15 μm. The influence of capillary dimensions and buffer composition on separation efficiency and sensitivity are discussed. Optimal separations were achieved using a 28 cm × 15 μm capillary, a separation high voltage of 35 kV, a background electrolyte of 25 mM ammonium acetate adjusted to pH 8.5, and negative ionization mode. The optimized method was successfully applied to a bovine testicular hyaluronidase digest of hyaluronan. Only minimal sample pretreatment for protein-containing samples is required. The simple manual injection procedure and fast separations allow for a sample throughput of 35 samples per hour.

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Abbreviations

BGE:

Background electrolyte

BSA:

Bovine serum albumin

BTH:

Bovine testicular hyaluronidase

CE:

Capillary electrophoresis

EOF:

Electroosmotic flow

ESI:

Electrospray ionization

ID:

Inner diameter

MS:

Mass spectrometry

NH4OAc:

Ammonium acetate

TOF-MS:

Time-of-flight mass spectrometry

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Acknowledgments

Financial support by the Deutsche Forschungsgemeinschaft (MA 1401/7-1) and the Bayerische Forschungsstiftung (AZ-829-08) is gratefully acknowledged.

This research was supported by the Research Executive Agency (REA) of the European Union under Grant Agreement number PITN-GA-2010-264772 (ITN CHEBANA).

MR expresses his gratitude to the Studienstiftung des Deutschen Volkes for a PhD grant.

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Authors and Affiliations

  1. Institute of Analytical Chemistry, Chemo- and Biosensors, University of Regensburg, 93040, Regensburg, Germany

    Marco Grundmann & Frank-Michael Matysik

  2. Institute of Pharmacy, University of Regensburg, 93040, Regensburg, Germany

    Martin Rothenhöfer, Günther Bernhardt & Armin Buschauer

Authors
  1. Marco Grundmann
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  2. Martin Rothenhöfer
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  3. Günther Bernhardt
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  4. Armin Buschauer
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  5. Frank-Michael Matysik
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Corresponding author

Correspondence to Frank-Michael Matysik.

Additional information

Published in the ANAKON special issue with guest editors P. Dittrich, D. Günther, G. Hopfgartner, and R. Zenobi

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Grundmann, M., Rothenhöfer, M., Bernhardt, G. et al. Fast counter-electroosmotic capillary electrophoresis–time-of-flight mass spectrometry of hyaluronan oligosaccharides. Anal Bioanal Chem 402, 2617–2623 (2012). https://doi.org/10.1007/s00216-011-5254-2

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  • DOI: https://doi.org/10.1007/s00216-011-5254-2

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