Increasing peak capacity in preparative liquid chromatography for pharmaceutical drug discovery using elevated temperature and column downsizing.

Preparative reversed-phase liquid chromatography remains a cornerstone technology for rapid, scalable purification of small-molecule pharmaceutical compounds providing easy access to high-purity compounds for structure-activity relationship studies, lead optimization, and early developability assessments. While temperature control is rarely optimized in preparative workflows, this study demonstrates that integrating elevated column and solvent temperatures with column downsizing (to 19 mm ID) doubles peak capacity, maintains large sample loading (up to 200 mg), and reduces solvent consumption by more than half, all with robust separation quality. Key operational strategies include dynamic heating to prevent temperature gradients and maximize reproducibility. Together, these optimizations provide an efficient, scalable, and cost-effective approach to compound purification in pharmaceutical drug discovery.