Optimization strategies for the analysis and purification of drug discovery compounds by reversed-phase high-performance liquid chromatography with high-pH mobile phases

Careful selection of both high-pH mobile phase as well as organic modifier, was performed in order to develop and optimize HPLC conditions for the separation of drug discovery compounds. High-pH mobile phases provide excellent chromatographic resolution and increased mass loading of basic compounds. The analytical methods so defined have been successfully transferred to preparative automated UV-directed purification, an important fact due to the increasing number of samples requiring purification. It should be noted that, the single prerequisite for this approach is an analytical LC–UV–MS run, therefore the system has the ability to collect only fractions likely to contain the target product. A cost-effective strategy for maximizing the purification of drug discovery compounds is proposed.