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

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.

Keywords

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

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

Supplementary material

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

References

  1. 1.
  2. 2.
    Chen WY, Abatangelo G (1999) Wound Repair Regen 7:79–89CrossRefGoogle Scholar
  3. 3.
    Noble PW (2002) Matrix Biol 21:25–29CrossRefGoogle Scholar
  4. 4.
    Day AJ, De la Motte CA (2005) Trends Immunol 26:637–643CrossRefGoogle Scholar
  5. 5.
    Boregowda RK, Appaiah HN, Siddaiah M, Kumarswamy SB, Sunila S, Thimmaiah KN, Mortha K, Toole B, Banerjee S (2006) J Carcinog 5:2–10CrossRefGoogle Scholar
  6. 6.
    Stern R (2005) Pathol Biol 53:372–382CrossRefGoogle Scholar
  7. 7.
    Stern R (2008) Semin Cancer Biol 18:275–280CrossRefGoogle Scholar
  8. 8.
    Grimshaw J, Kane A, Trocha-Grimshaw J, Douglas A, Chakravarthy U, Archer D (1994) Electrophoresis 15:936–940CrossRefGoogle Scholar
  9. 9.
    Grimshaw J, Trocha-Grimshaw J, Fisher W, Rice A, Smith S, Spedding P, Duffy J, Mollan R (1996) Electrophoresis 17:396–400CrossRefGoogle Scholar
  10. 10.
    Hofinger ESA, Bernhardt G, Buschauer A (2007) Glycobiology 17:963–971CrossRefGoogle Scholar
  11. 11.
    Hofinger ESA, Hoechstetter J, Oettl M, Bernhardt G, Buschauer A (2008) Glycoconj J 25:101–109CrossRefGoogle Scholar
  12. 12.
    Hjerten S, Valtcheva L, Elenbring K, Liao JL (1995) Electrophoresis 16:584–594CrossRefGoogle Scholar
  13. 13.
    Hutterer KM, Jorgenson JW (1999) Anal Chem 71:1293–1297CrossRefGoogle Scholar
  14. 14.
    Palonen S, Jussila M, Porras SP, Hyötyläinen T, Riekkola ML (2001) J Chromatogr A 916:89–99CrossRefGoogle Scholar
  15. 15.
    Palonen S, Jussila M, Porras SP, Hyötyläinen T, Riekkola ML (2002) Electrophoresis 23:393–399CrossRefGoogle Scholar
  16. 16.
    Zhong M, Lunte S (1996) Anal Chem 68:2488–2493CrossRefGoogle Scholar
  17. 17.
    Müller O, Minarik M, Foret F (1998) Electrophoresis 19:1436–1444CrossRefGoogle Scholar
  18. 18.
    Matysik FM (2006) Electrochem Commun 8:1011–1015CrossRefGoogle Scholar
  19. 19.
    Matysik FM, Neusüß C, Pelzing M (2008) Analyst 133:1764–1766CrossRefGoogle Scholar
  20. 20.
    Opekar F, Coufal P, Štulík K (2009) Chem Rev 109:4487–4499CrossRefGoogle Scholar
  21. 21.
    Kennedy RT (1999) Anal Chim Acta 400:163–180CrossRefGoogle Scholar
  22. 22.
    Wallingford RA, Ewing AG (1988) Anal Chem 60:1975–1977CrossRefGoogle Scholar
  23. 23.
    Olefirowicz TM, Ewing AG (1990) Anal Chem 62:1872–1876CrossRefGoogle Scholar
  24. 24.
    Chien JB, Wallingford RA, Ewing AG (1990) J Neurochem 54:633–638CrossRefGoogle Scholar
  25. 25.
    Olefirowicz TM, Ewing AG (1990) J Neurosci Meth 34:11–15CrossRefGoogle Scholar
  26. 26.
    Kristensen HK, Lau YY, Ewing AG (1994) J Neurosci Meth 51:183–188CrossRefGoogle Scholar
  27. 27.
    Kennedy RT, Oates MD, Cooper BR, Nickerson B, Jorgenson JW (1989) Science 246:57–63CrossRefGoogle Scholar
  28. 28.
    Hogan BL, Yeung ES (1992) Anal Chem 64:2841–2845CrossRefGoogle Scholar
  29. 29.
    Blasco S, Kortz L, Matysik FM (2009) Electrophoresis 30:1–6CrossRefGoogle Scholar
  30. 30.
    Lin Y, Trouillon R, Safina G, Ewing AG (2011) Anal Chem 83:4369–4392CrossRefGoogle Scholar
  31. 31.
    Lapainis T, Rubakhin SS, Sweedler JV (2009) Anal Chem 81:5858–5864CrossRefGoogle Scholar
  32. 32.
    Grundmann M, Matysik FM (2011) Anal Bioanal Chem 400:269–278CrossRefGoogle Scholar
  33. 33.
    König S, Fales HM (1999) J Am Soc Mass Spectrom 10:273–276CrossRefGoogle Scholar
  34. 34.
    Pittman JL, Schrum KF, Gilman SD (2001) Analyst 126:1240–1247CrossRefGoogle Scholar
  35. 35.
    Reissig JL, Strominger JL, Leloir LF (1955) J Biol Chem 217:959–966Google Scholar
  36. 36.
    Muckenschnabel I, Bernhardt G, Spruss T, Dietl B, Buschauer A (1998) Cancer Lett 131:13–20CrossRefGoogle Scholar
  37. 37.
    Hascall VC, Laurent TC (1997) Glycoforum. http://www.glycoforum.gr.jp/science/hyaluronan/HA01/HA01E.html

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

Personalised recommendations