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On-column solvent exchange for purified preparative fractions
Gilles H Goetz1, Emily Beck, Peter W Tidswell
1Pfizer Global Research and Development, Analytical Chemistry and Sample Logistics, St Louis Laboratories, Chesterfield, MO, USA. gilles.h.goetz@pfizer.com
Journal of Laboratory Automation
|September 13, 2011
Summary
On-column solvent exchange significantly reduces evaporation time for preparative HPLC fractions. This method improves sample purity and reduces risks of salt formation and decomposition.
Area of Science:
- Analytical Chemistry
- Chromatography
Background:
- Preparative High-Performance Liquid Chromatography (HPLC) often yields fractions in large volumes of solvent mixtures, typically acetonitrile:water.
- Evaporation of these large-volume aqueous fractions is time-consuming and can lead to sample degradation.
Purpose of the Study:
- To implement an on-column solvent exchange technique to reduce evaporation cycle times for preparative HPLC fractions.
- To assess the impact of this method on sample purity, recovery, and potential for degradation.
Main Methods:
- Utilized principles of solid-phase extraction for on-column solvent exchange within the preparative HPLC system.
- Injected acetonitrile:water fractions and re-eluted in a volatile organic solvent, primarily acetonitrile.
- Evaluated evaporation times, sample recovery, purity, and potential for salt formation and thermal decomposition.
Main Results:
- Reduced acetonitrile evaporation time from 17 hours for aqueous fractions to 3 hours.
- Enabled the use of lower temperatures during the evaporation step, minimizing thermal decomposition.
- Effectively removed mobile-phase additives that could form salts, improving sample stability.
- Demonstrated no adverse effects on sample recovery or purity.
Conclusions:
- On-column solvent exchange is an efficient method to accelerate the evaporation of preparative HPLC fractions.
- This technique offers significant advantages including reduced cycle times, minimized sample degradation, and improved purity.
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