Skip to main content
SpringerLink
Log in

Optimal conditions for the reversed-phase chromatography of proteins

  • Published:
Chromatographia Aims and scope Submit manuscript

Summary

Optimum stationary phases for reversed phase chromatography should be based on a silica with a large average pore diameter, preferably above 30nm. To exclude anormalous elution behaviour of proteins, octadecyl groups should be bonded to the surface and the residual silanol groups should be reacted with a highly active silanization agent like bistrimethylsilyl acetamide. The total amount of bonded carbon should not exceed 5% w/w. Because of the low diffusion coefficients of proteins, the particle diameter should be as small as possible. Protein retention can hardly be influenced by changes of the organic modifier or by temperature. Because of solubility and viscosity, however, acetonitrile is to be prefered.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. B. H. J. Hofstee, in “Methods of Protein Separation”,N. Catsimpoolas ed., Vol. 2, p. 233, Plenum, New York (1976).

    Google Scholar 

  2. W. S. Hancock, J. T. Sparrow, in “High-Performance Liquid Chromatography, Advances and Perspectives”,Cs. Horváth ed., Vol. 3, p. 49–85, Academic Press, New York (1983).

    Google Scholar 

  3. M. J. O’Hare, E. C. Nice, J. Chromatogr.171, 209 (1979).

    Article  CAS  Google Scholar 

  4. N. H. C. Cooke, B. G. Archer, M. J. O’Hare, E. C. Nice, M. Capp, J. Chromatogr.255, 115 (1983).

    Article  CAS  Google Scholar 

  5. R. V. Lewis, A. Fallon, S. Stein, K. D. Gibson, S. Udenfriend, Anal. Biochem.104, 153 (1980).

    Article  CAS  Google Scholar 

  6. J. E. Rivier, J. Liq. Chromatogr.1, 343 (1978).

    Article  CAS  Google Scholar 

  7. S. Terabe, H. Nishi, T. Ando, J. Chromatogr.212, 293 (1981).

    Article  Google Scholar 

  8. E. C. Nice, M. W. Capp, N. Cooke, M. J. O’Hare, J. Chromatogr.218, 569 (1981).

    Article  CAS  Google Scholar 

  9. Cs. Horváth, W. R. Melander, I. Mołnár, J. Chromatogr.125, 129 (1976).

    Article  Google Scholar 

  10. Cs. Horváth, W. R. Melander, J. Chromatogr. Sci.15, 393 (1977).

    Google Scholar 

  11. J. B. Shelton, J. R. Shelton, W. A. Schroeder, Hemoglobin3, 353 (1979).

    CAS  Google Scholar 

  12. J. D. Pearson, W. C. Mathoney, M. A. Hermodson, F. R. Regnier, J. Chromatogr.207, 325 (1981).

    Article  CAS  Google Scholar 

  13. J. D. Pearson, N. T. Lin, F. R. Regnier, Anal. Biochem.124, 217 (1982).

    Article  CAS  Google Scholar 

  14. R. A. Barford, B. J. Sliwinski, A. C. Breyer, H. L. Rothbart, J. Chromatogr.235, 281 (1982).

    Article  CAS  Google Scholar 

  15. H. Engelhardt, “High-Performance Liquid Chromatography”, Springer, Berlin, Heidelberg, New York (1979).

    Google Scholar 

  16. W. R. Melander, Cs. Horváth, in “High-Performance Liquid Chromatography, Advances in Perspectives”, Vol. 2, p. 113,Cs. Horváth ed., Academic Press, New York (1980).

    Google Scholar 

  17. J. C. Giddings, J. Gas Chromatography5, 413 (1967).

    Google Scholar 

  18. M. T. W. Hearn, B. Grego, J. Chromatogr.218, 497 (1981).

    Article  CAS  Google Scholar 

  19. B. L. Karger, J. R. Grant, A. Hartkopf, P. W. Weiner, J. Chromatogr.128, 65 (1976).

    Article  CAS  Google Scholar 

  20. W. R. Melander, A. Nahum, Cs. Horváth, J. Chromatogr.185, 129 (1979).

    Article  CAS  Google Scholar 

  21. A. Nahum, Cs. Horváth, J. Chromatogr.203, 53 (1981).

    Article  CAS  Google Scholar 

  22. K. E. Bij, Cs. Horváth, W. R. Melander, A. Nahum, J. Chromatogr.203, 65 (1981).

    Article  CAS  Google Scholar 

  23. H. El gass, H. Engelhardt, I. Halász, Z. Anal. Chem.294, 97 (1979).

    Article  Google Scholar 

  24. I. Halász, G. Maldener, Anal. Chem.,55, 1842 (1983).

    Article  Google Scholar 

  25. H. Engelhardt, D. Mathes, J. Chromatogr.185, 305 (1979).

    Article  CAS  Google Scholar 

  26. H. Engelhardt, G. Ahr, Chromatographia14, 227 (1981).

    Article  CAS  Google Scholar 

  27. H. Engelhardt, B. Dreyer, H. Schmidt, Chromatographia16, 11 (1982).

    Article  CAS  Google Scholar 

  28. I. Halász, K. Martin, Angew. Chem.90, 954 (1978).

    Article  Google Scholar 

  29. H. Engelhardt, H. Müller, J. Chromatography218, 395 (1981).

    Article  CAS  Google Scholar 

  30. L. R. Snyder, in “High-Performance Liquid Chromatography, Advances and Perspectives”, Vol. 1, p. 207,Cs. Horváth ed., Academic Press, New York (1980).

    Google Scholar 

  31. K. K. Unger inW. S. Hancock ed., “Handbook of HPLC for the Separation of Amino Acids, Peptides and Proteins”, CRC Press Inc., Boca Raton, Flo., (1984).

    Google Scholar 

  32. S. A. Cohen, K. P. Benedek, S. Dong, Y. Tapuhi, B. L. Karger, Anal. Chem.,56, 217 (1984).

    Article  CAS  Google Scholar 

  33. J. D. Pearson, F. E. Regnier, J. Liquid Chromatogr.6, 496 (1983).

    Google Scholar 

  34. L. R. Snyder, J. W. Dolan, J. R. Grant, J. Chromatogr.165, 3 (1979).

    Article  CAS  Google Scholar 

  35. P. E. Petrides, R. T. Jones, P. Bohlen, Anal. Biochem.105, 383 (1980).

    Article  CAS  Google Scholar 

  36. S. Terabe, H. Nishi, T. Ando, J. Chromatogr.212, 295 (1981).

    Article  CAS  Google Scholar 

  37. M. J. O’Hare, M. W. Capp, E. C. Nice, N. H. C. Cooke, B. G. Archer, Anal. Biochem.126, 17 (1982).

    Article  CAS  Google Scholar 

  38. S. A. Cohen, K. Schellenberg, K. Benedek, B. L. Karger, B. Grego, M. T. W. Hearn, Anal. Biochem.140, 223 (1984).

    Article  CAS  Google Scholar 

  39. Ph. D. ThesisU. M. Schön, Saarbrücken 1984.

Download references

Author information

Authors and Affiliations

  1. Angewandte Physikalische Chemie, Universität des Saarlandes, Saarbrücken, FRG

    H. Engelhardt & H. Müller

Authors
  1. H. Engelhardt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H. Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Engelhardt, H., Müller, H. Optimal conditions for the reversed-phase chromatography of proteins. Chromatographia 19, 77–84 (1984). https://doi.org/10.1007/BF02687722

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02687722

Key Words

Search

Navigation