Analytical and Bioanalytical Chemistry

, Volume 402, Issue 8, pp 2617–2623 | Cite as

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

  • Marco Grundmann
  • Martin Rothenhöfer
  • Günther Bernhardt
  • Armin Buschauer
  • Frank-Michael Matysik
Original Paper


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.


Capillary electrophoresis Counter-electroosmotic Time-of-flight mass spectrometry High electric field strength Hyaluronan Bovine testicular hyaluronidase 



Background electrolyte


Bovine serum albumin


Bovine testicular hyaluronidase


Capillary electrophoresis


Electroosmotic flow


Electrospray ionization


Inner diameter


Mass spectrometry


Ammonium acetate


Time-of-flight mass spectrometry



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.

Supplementary material

216_2011_5254_MOESM1_ESM.pdf (170 kb)
ESM 1 (PDF 170 kb)


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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Marco Grundmann
    • 1
  • Martin Rothenhöfer
    • 2
  • Günther Bernhardt
    • 2
  • Armin Buschauer
    • 2
  • Frank-Michael Matysik
    • 1
  1. 1.Institute of Analytical Chemistry, Chemo- and BiosensorsUniversity of RegensburgRegensburgGermany
  2. 2.Institute of PharmacyUniversity of RegensburgRegensburgGermany

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