, Volume 14, Issue 10, pp 589–592 | Cite as

Role of the organic solvent modifier in the reversed phase high-performance liquid chromatography of polypeptides

  • B. Grego
  • M. T. W. Hearn


The retention behaviour of several polypeptide hormones on μBondapak C18 columns has been examined using mobile phases of different acetonitrile percentage compositions containing 15 mM triethylammonium phosphate or 15 mM orthophosphoric acid buffers. Under these elution conditions, the capacity factors and the selectivity parameters of these polypeptide hormones show pronounced dependencies on the volume fraction of the organic solvent modifier. In the range 0–40% acetonitrile, the capacity factors were monotonously attenuated with increasing modifier percentages with the elution order essentially in accord with that hat anticipated for a reversed phase separation mode. At higher concentrations of acetonitrile, retention of the polypeptides to the octadecylsilica support progressively increased with elution order reversals indicative of a normal- or polarphase separation mode. These observations are discussed in terms of the interplay of hydrophobic and silanophilic interactions which occur between the ionised polypeptides and the stationary phase under these changing mobile phase conditions.

Key Words

High-performance liquid chromatography Polypeptides Solvent modifier 


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  1. [1]
    H. Engelhardt, G. Ahr andM. T. W. Hearn, J. Liquid Chromatogr. (in press.).Google Scholar
  2. [2]
    M. T. W. Hearn andB. Grego, J. Chromatogr.203, 349 (1981).CrossRefGoogle Scholar
  3. [3]
    M. T. W. Hearn, B. Grego andW. S. Hancock, J. Chromatogr.185, 429 (1979).CrossRefPubMedGoogle Scholar
  4. [4]
    M. T. W. Hearn, J. Liquid Chromatogr.3, 1255 (1980).Google Scholar
  5. [5]
    M. T. W. Hearn, in HPLC: Advances and Perspectives (Cs. Horvath, ed.), Academic Press, New York, N.Y. vol. 3, in press.Google Scholar
  6. [6]
    M. T. W. Hearn andW. S. Hancock, in Biological/Biomedical Applications of Liquid Chromatography (G. L. Hawk, ed.). Marcel Dekker, New York, N.Y., 1979, pp. 243–271.Google Scholar
  7. [7]
    M. T. W. Hearn, in Advances in Chromatography (J. C. Giddings, E. Grushka, J. Cazes andP. Brown, eds.), Marcel Dekker, Inc., New York, N.Y., Vol. 18, 1980, pp. 59–100.Google Scholar
  8. [8]
    M. J. O'Hare andE. C. Nice, J. Chromatogr.171, 209 (1979).CrossRefPubMedGoogle Scholar
  9. [9]
    M. T. W. Hearn, B. Grego andG. Chapman, submitted for publication.Google Scholar
  10. [10]
    E. C. Nice, M. Capp andM. J. O'Hare, J. Chromatogr.185, 413 (1979).CrossRefPubMedGoogle Scholar
  11. [11]
    M. T. W. Hearn andB. Grego, J. Liquid Chromatogr., in press.Google Scholar
  12. [12]
    W. Melander andCs. Horvath, in HPLC: Advances and Perspectives (Cs. Horvath, ed.), Academic Press, New York, N. Y., 1980, Vol. 2, pp. 113–319.Google Scholar
  13. [13]
    A. Nahum, W. R. Melander andCs. Horvath, J. Chromatogr. in press.Google Scholar
  14. [14]
    C. Hunter, K. Sugden andJ. G. Lloyd-Jones, J. Liquid Chromatogr.3, 1335 (1980).Google Scholar
  15. [15]
    M. T. W. Hearn andB. Grego, J. Chromatogr., submitted for publication.Google Scholar
  16. [16]
    Cs. Horvath, W. Melander andI. Molnar, J. Chromatogr.125, 129 (1976).CrossRefGoogle Scholar
  17. [17]
    K. K. Unger, in Porous Silica, Elsevier Scientific Publ. Co., Amsterdam, 1979, p. 214.Google Scholar
  18. [18]
    R. P. W. Scott andP. Kucera, J. Chromatogr.175, 51 (1979).CrossRefGoogle Scholar
  19. [19]
    J. H. Knox andA. Pryde, J. Chromatogr.112, 171 (1975).CrossRefGoogle Scholar
  20. [20]
    A. Sokolowski andK. G. Wahlund, J. Chromatogr.189, 299 (1980).CrossRefGoogle Scholar
  21. [21]
    W. R. Melander, J. Stoveken andC. Horvath, J. Chromatogr.185, 111 (1979).CrossRefGoogle Scholar
  22. [22]
    J. Rivier, J. Liquid Chromatogr.1, 347 (1978).Google Scholar
  23. [23]
    W. S. Hancock, C. A. Bishop, J. E. Battersby, D. R. K. Harding andM. T. W. Hearn, J. Chromatogr.168, 377 (1979).CrossRefGoogle Scholar
  24. [24]
    J. G. R. Hurrell, R. J. Fleming andM. T. W. Hearn, J. Liquid Chromatogr.3, 473 (1980).Google Scholar
  25. [25]
    M. T. W. Hearn, B. Grego, P. Stanton andA. J. Paterson. Manuscript in preparation.Google Scholar
  26. [26]
    M. T. W. Hearn, Proc. Endocr. Soc. Austr.23, 21 (1980).Google Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1981

Authors and Affiliations

  • B. Grego
    • 1
  • M. T. W. Hearn
    • 1
  1. 1.St. Vincent's School of Medical ResearchMelbourneAustralia

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