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Updated: May 22, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
Screening strategy for chiral and achiral separations in supercritical fluid chromatography mode
David Speybrouck1, David Corens, Jean-Michel Argoullon
1Research Department Janssen France, Campus de Maigremont 27106 Val de Reuil cedex France. dspeybroucki@its.jnj.com
Abstract:
Supercritical fluid (SF) was discovered 200 years ago, but the use of this fluid as a mobile phase in chromatography only became popular fifty years ago. The development of the supercritical fluid chromatography (SFC) was progressing slowly due to technological problems since ten years; the interest for this chromatographic mode has been growing up as the construction of the SFC instruments is more or less similar with HPLC instruments. The main difference in SFC is the installation of a back pressure regulator which is implemented to control the pressure above the critical pressure. SFC is widely used in chiral chromatography where Polysaccharide phases are the most versatile in use. The mobile phase is mainly composed by CO(2) but the polarity can be increased by adding alcohol. The nature of the alcohol can change drastically the selectivity. The choice of the best tandem stationary phase / mobile phase is difficult to predict. Hence a full screening with different stationary phases and mobile phase solvents is often mandatory. For the achiral separation, SFC is more and more used. Achiral SFC can be classified as normal phase mode, it means that stationary phases are more polar than mobile phase and retention times decrease as polarity of the mobile phase increases. Most popular stationary phases are silica linked with polar group such as aminopropyl, cyanoprpyl, diol or 2-ethylpyridine. Mobile phase are generally composed by CO(2) and methanol. SFC can be used as a complementary technique for reversed phase HPLC or sometimes even to replace HPLC.
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