Separation of negatively charged isomeric quinones in acidic solution by capillary electrophoresis with reductive electrochemical detection

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Amy R. Smith, Jon R. Kirchhoff, Richard A. Hudson, L. M. V. Tillekeratne and Richard A. Hudson


Abstract

A capillary electrophoresis method with reductive electrochemical detection was developed for the separation of the novel enzyme cofactor pyrroloquinoline quinone (PQQ) and three isomeric analogues. Tuning the reduction potential of the o-quinone moiety to a value more positive than the reduction potential for oxygen was accomplished by adjusting the capillary buffer to pH 2, thus eliminating the need for deoxygenation. To counter the suppression of the electroosmotic flow (EOF) at pH 2, a negative separation voltage of –22.5 V was applied to a 25 µm id capillary resulting in migration of the anionic isomers toward the electrochemical detector. Fast and efficient separation was achieved in 0.15 mM phosphate buffer at pH 2. A mass detection limit for PQQ of 2 fmol was obtained with end-column detection. This approach may find utility for the separation and sensitive detection of a wide range of reducible quinones.


References

  1. S. A. Salisbury, H. S. Forrest, W. B. T. Cruse and O. Kennard, Nature, 1979, 280, 843 CAS.
  2. J. A. Duine and J. Frank, FEMS Microbiol. Rev., 1990, 87, 221 CrossRef CAS.
  3. C. Hartmann and J. P. Klinman, Biofactors, 1988, 1, 41 Search PubMed.
  4. J. A. Duine, J. Frank, Jr. and J. K. Van Zeeland, FEBS Lett., 1979, 108, 443 CrossRef CAS.
  5. Y. Hamagishi, S. Murata, H. Kamie, T. Oki, O. Adachi and M. Ameyama, J. Pharmacol. Exp. Ther., 1990, 255, 980 Search PubMed.
  6. F. Xu, C. P. Mack, K. S. Quandt, M. Shlafer, V. Massey and D. E. Hultquist, Biochem. Biophys. Res. Commun., 1993, 193, 434 CrossRef CAS.
  7. K. S. Quandt and D. E. Hultquist, Proc. Natl. Acad. Sci., 1994, 91, 9322 CAS.
  8. A. Bishop, P. M. Gallop and M. L. Karnovsky, Nutr. Rev., 1998, 56, 287 Search PubMed.
  9. B. W. Groen, R. A. van der Meer and J. A. Duine, FEBS Lett., 1988, 237, 98 CrossRef CAS.
  10. E. Aizenman, F. E. Jensen, P. M. Gallop, P. A. Rosenberg and L. H. Tang, Neurosci. Lett., 1994, 168, 189 CAS.
  11. J. P. Klinman, D. M. Dooley, J. A. Duine, P. F. Knowles, B. Mondovi and J. J. Villafranca, FEBS Lett., 1991, 282, 1 CrossRef CAS.
  12. W. S. McIntire, D. E. Wemmer, A. Christoserdov and M. E. Lidstrom, Science, 1991, 252, 817 CrossRef CAS.
  13. E. D. Harris, Nutr. Rev., 1992, 50, 263 Search PubMed.
  14. L. A. Holland and S. M. Lunte, Anal. Commun., 1998, 35, 1H RSC.
  15. P. D. Voegel and R. P. Baldwin, Electrophoresis, 1997, 18, 2267 CAS.
  16. A. G. Ewing, J. M. Mesaros and P. F. Gavin, Anal. Chem., 1994, 66, 527A CAS.
  17. Y. Esaka, Y. Yamaguchi, K. Kano and M. Goto, J. Chromatogr. A, 1993, 652, 225 CrossRef CAS.
  18. Y. Esaka, Y. Yamaguchi, M. Goto and K. Kano, J. Chem. Soc., Perkin Trans. 2, 1994, 2163 RSC.
  19. A. R. Smith, J. R. Kirchhoff, Z. Zhang, L. M. V. Tillekeratne and R. A. Hudson, J. Chromatogr. A, submitted Search PubMed.
  20. J. P. Klinman and K. Mu, Annu. Rev. Biochem., 1994, 63, 299 CrossRef CAS.
  21. P. R. Bergethon, Anal. Biochem., 1990, 186, 324 CAS.
  22. Y. Esaka, K. Kano, M. Sukeguchi and M. Goto, Anal. Sci., 1993, 9, 207 CAS.
  23. M. Otani, C. Umezawa and K. Sano, J. Chromatogr., 1989, 483, 448 CrossRef CAS.
  24. Z. Zhang, L. M. V. Tillekeratne, J. R. Kirchhoff and R. A. Hudson, Biochem. Biophys. Res. Commun., 1995, 212, 41 CrossRef CAS.
  25. K. Kano, K. Mori, B. Uno, T. Kubota, T. Ikeda and M. Senda, Bioelectrochem. Bioenerg., 1990, 24, 193 CrossRef CAS.
  26. K. Kano, K. Mori, B. Uno, T. Kubota, T. Ikeda and M. Senda, Bioelectrochem. Bioenerg., 1990, 23, 227 CrossRef CAS.
  27. K. D. Lukcas and J. W. Jorgenson, J. High Resolut. Chromatogr., Chromatogr. Commun., 1985, 8, 407 Search PubMed.
  28. Z. Zhang, L. M. V. Tillekeratne and R. A. Hudson, Synthesis, 1996, 377 CrossRef CAS.
  29. T. J. O'Shea, R. D. Greenhagen, S. M. Lunte, C. E. Lunte, M. R. Smyth, D. M. Radzik and N. Wantanabe, J. Chromatogr. A, 1992, 593, 305 CrossRef CAS.
  30. X. Huang, R. N. Zare, S. Sloss and A. G. Ewing, Anal. Chem., 1991, 63, 189 CrossRef CAS.
  31. S. R. Wallenborg, L. Nyholm and C. E. Lunte, Anal. Chem., 1999, 71, 544 CrossRef CAS.
  32. M. A. Malone, P. L. Weber, M. R. Smyth and S. M. Lunte, Anal. Chem., 1994, 66, 3782 CrossRef CAS.
  33. R. M. McCormick, Anal. Chem., 1988, 60, 2322 CrossRef CAS.
  34. A. J. Mort and E. M. V. Chen, Electrophoresis, 1996, 17, 379 CAS.
  35. G. M. Janini, G. M. Muschik and H. J. Issaq, J. Chromatogr. B, 1996, 683, 29 CrossRef CAS.
  36. Y. Y. Dong, Y. Sun and Z. Sun, J. High Resolut. Chromatogr., 1998, 21, 445 CrossRef CAS.
  37. W. Ding and J. S. Fritz, Anal. Chem., 1998, 70, 1859 CrossRef CAS.
  38. Y. He and H. K. Lee, Anal. Chem., 1998, 71, 995 CrossRef CAS.
  39. M. J. Thronton and J. S. Fritz, J. Chromatogr. A, 1997, 770, 301 CrossRef CAS.
  40. H. T. Chang, H. S. Chen and R. Lee, J. Chromatogr. A, 1998, 800, 339 CrossRef CAS.
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