Chromatographia

, 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
Originals

Summary

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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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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