High speed gradient elution reversed-phase liquid chromatography

Adam P. Schellinger, Dwight R. Stoll, Peter W. Carr

Research output: Contribution to journalArticlepeer-review

84 Scopus citations


A major disadvantage of gradient elution in terms of speed results from the need to adequately re-equilibrate the column. This work distinguishes two states of re-equilibration: (1) run-to-run repeatability and (2) full equilibration. We find that excellent repeatability (±0.002 min in retention time) is achieved with at most 2 column volumes of re-equilibration whereas full equilibration can require considerably more than 20 column volumes. We have investigated the effects of adding ancillary solvents (e.g. n-propanol, n-butanol) to the eluent and changing the particle pore size, initial eluent composition and type, column temperature and flow rate on the speed of full equilibration. Full equilibration seems to be more thermodynamically limited than kinetically controlled. Also, we show that the main limitation to reducing the full equilibration time is related to instrument design issues; a novel approach to overcome these instrumental issues is described.

Original languageEnglish (US)
Pages (from-to)143-156
Number of pages14
JournalJournal of Chromatography A
Issue number2
StatePublished - Feb 4 2005

Bibliographical note

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


  • Equilibration
  • Flush-out volume
  • Gradient elution
  • Speed


Dive into the research topics of 'High speed gradient elution reversed-phase liquid chromatography'. Together they form a unique fingerprint.

Cite this