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.