Molded separation media: An inexpensive, efficient, and versatile alternative to packed columns for the fast HPLC separation of peptides, proteins, and synthetic oligomers and polymers

Abstract A simple molding process carried out within the confines of a chromatographic column has been used for the preparation of macroporous poly(glycidyl methacrylate‐ co ‐ethylene dimethacrylate) and poly(styrene‐ co ‐divinylbenzene) rods. The novel monolithic separation media that are obtained are useful for the HPLC separation of biological and synthetic polymers. The presence of large pores with a diameter of about 1 μm makes the molded rod columns easily permeable to eluents. Therefore, the back pressure of these columns is modest even at high flow rates. In contrast to the conventional HPLC columns packed with beads, all of the mobile phase flows through the continuous monolithic medium. As a result of this total convection, the efficiency of the molded media is almost independent of the flow rate. This improves significantly the separation ability of the rod columns and very fast separations of macromolecules such as peptides, proteins, and synthetic polymers have been demonstrated.