Systematic method development in hydrophobic interaction chromatography I. Characterization of the phase system and modelling retention

Ge ́za Rippel, A. ́gota Bede, La ́szlo ́. Szepesy

Research output: Contribution to journalArticle

10 Citations (Scopus)


In order to rationalize the selection of the phase system (the stationary phase and the mobile phase) applicable for the separation of some proteins, gradient measurements were carried out on three columns of different types using three salts as eluent constituents. The slope and intercept of the lnk-salt concentration relationship for proteins varied with the different phase systems. The hydrophobicity index (c*) showed how the alteration of the type of phase system affects the selectivity of separation. Two-dimensional mapping along the parameters of the slope-intercept relationship revealed the extent of the difference among the phase systems. In addition, the effect of solutions of binary and ternary salt mixtures on the retention of proteins was investigated. Measurements were carried out according to a mixture design and the retention time was modelled with the predictive method of Jandera and by fitting a polynomial to the data according to the Sentinel method. The accuracy of the former method were dependent on the type of phase system. The fit of second-order polynomial gave very good results but a first-order model was also acceptable.

Original languageEnglish
Pages (from-to)17-29
Number of pages13
JournalJournal of Chromatography A
Issue number1-2
Publication statusPublished - Apr 21 1995


ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this