A practical approach to transferring linear gradient elution methods

Adam P. Schellinger, Peter W. Carr

Research output: Contribution to journalArticlepeer-review

50 Scopus citations


Attempts to theoretically address the problems involved in transferring linear gradient elution methods have been somewhat ad hoc due to the simplifying assumptions usually made in conventional gradient elution theory. Until now, all equations based on the k* parameter of linear gradient elution theory used as the basis for predicting the separation selectivity have not explicitly included the effect of the dwell volume (VD). Using an exact equation for predicting k*, that is, one which fully accounts in an a priori fashion for VD, we find a set of simple yet exact equations which unequivocally must be satisfied to transfer an optimized linear gradient elution method from one system (column or instrument or both) to another. These relationships absolutely mandate that a change in the instrument dwell volume requires a proportional change in the column volume; in turn, a change in the column volume requires a proportional change in the flow rate and/or gradient time to maintain a constant gradient steepness. Although we are not the first to suggest these guidelines, this work provides a complete theoretical foundation for these exact guidelines for the maintenance of gradient selectivity for the case of transferring a linear gradient elution method between different columns packed with the same particles and/or between different instruments.

Original languageEnglish (US)
Pages (from-to)110-119
Number of pages10
JournalJournal of Chromatography A
Issue number2
StatePublished - Jun 10 2005

Bibliographical note

Funding Information:
The authors acknowledge financial support from the National Institutes of Health (Grant # 5R01GM054585-09).


  • Dwell volume
  • Gradient elution
  • Method transfer
  • Selectivity


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