New insights into supercritical fluid chromatography for chiral separations

Abstract

Bioanalysis of stereoisomers represents a great challenge, as chromatographic separation is not feasible using conventional reversed or normal-phase chromatography. Supercritical fluid chromatography in combination with chiral stationary phases has proven to be a great tool for chiral resolution, alleviating some of the challenges associated with bioanalysis of stereoisomers. This technical note describes an alternative to the traditional SFC instrument configuration without the use of a back pressure regulator (BPR) and makeup pump; thereby simplifying its usage and improving the robustness and quality of measurements. This alternative configuration has been successfully employed in our laboratory for multiple chiral applications; in particular for the separation of cis/trans geometric stereoisomers. Four case studies are presented – one of which was validated demonstrating the robustness of the technology for bioanalytical applications.