Development of an automated dual‐mode supercritical fluid chromatography and reversed‐phase liquid chromatography mass‐directed purification system for small‐molecule drug discovery

Abstract

We report the development of a dual-mode mass-directed supercritical fluid chromatography and reversed-phase liquid chromatography purification system. The addition of a third pump allows for flexible mobile phase control between the two techniques, and enables operation of either chromatography mode within minutes by activation of a set of switching valves on a single system. Software control, fluidic pathways, interface to the mass spectrometer, and fraction collection have been modified for compatibility between both separation methods. The conditioning solvent and tuning parameters for the mass spectrometer were adjusted to achieve an ideal signal trace in either mode with good linearity (r(2) > 0.970) over a range of concentrations and minimal noise for accurate peak detection and isolation. The registration success rate is 90% and overall sample recovery for either technique is 80-90%. Combining two orthogonal separation and purification modes in one single system has improved the purification throughput of complex mixtures and has been a valuable, cost-saving tool in our laboratory.

Use and practice of achiral and chiral supercritical fluid chromatography in pharmaceutical analysis and purification

“…Recently, there has been an increasing interest for achiral purification as well . In addition to the advantages previously mentioned, the alternative selectivity provided by SFC compared to HPLC provides an interesting separation tool for achiral high‐throughput purification .…”

Microscale Purification with Direct Charged Aerosol Detector Quantitation Using Selective Online One- or Two-Dimensional Liquid Chromatography

“…Automated mass-directed HPLC systems with fraction collection − are routinely employed for the separation and isolation of pharmaceuticals derived from parallel medicinal chemistry (PMC) techniques. Specialized groups in high-throughput experimentation (HTE) − and purification (HTP) − with dedicated separations capability are tasked with providing high-value services in support of discovery chemistry. A variety of chromatographic technologies for screening and purification − have been developed for these environments to meet increasing demand.…”

Efficient separation of curcumin, demethoxycurcumin, and bisdemethoxycurcumin from turmeric using supercritical fluid chromatography: From analytical to preparative scale

“…SFC is an environment‐friendly technique, which consumes far less organic solvents than HPLC . It also shows excellent orthogonality to RP‐LC . SFC has been successfully applied to both analytical and preparative scale separation of natural compounds with similar structures, such as steroids, terpenoids, and isoflavones .…”