High throughput analysis and purification in support of automated parallel synthesis.

Rapid reverse-phase analytical and preparative HPLC methods have been developed for application to parallel synthesis libraries. Gradient methods, short columns, and high flow rates allow analysis of over 300 compounds per day on a single system, or purification of up to 200 compounds per day on a single preparative system. Hardware and software modifications allow continuous unattended use for maximum efficiency and throughput.

Purification of synthetic peptides on a high-resolution preparative reversed-phase column.

A 2-in. I.D. column filled with 10-microns spherical C18 bonded silica with 120 A pores was used for the preparative purification of various synthetic peptides in one step. The small-sized silica packing afforded high resolution and the spherical shape helped maintain a relatively low back-pressure during the chromatography. Conditions for performing the separations were derived from the analytical chromatography of samples on a column of similar 5-microns material. The same amount of organic modifier, but with the gradient duration increased, achieved very similar separations on the preparative column.