Efficient polymer monolith for strong cation-exchange capillary liquid chromatography of peptides

A stable poly[ 2-acrylamido-2-methyl-1-propanesulfonic acid-co-poly( ethylene glycol) diacrylate] monolith was synthesized inside a 75-mu m-i.d. capillary by photoinitiated copolymerization with water, methanol, and ethyl ether as porogens. The resulting monolith was evaluated for strong cation-exchange capillary liquid chromatography of both synthetic and natural peptides. Although the monolith possessed relatively strong hydrophobicity due to the use of 2-acrylamido-2-methyl-1-propanesulfonic acid as one monomer, the monolith had a high dynamic binding capacity of 157 mu equiv of peptide/mL, or 332 mg of cytochrome c/mL. Exceptionally high resolution resulting from extremely narrow peaks was obtained, resulting in a peak capacity of 179 when using a shallow salt elution gradient. Although a second, naturally formed gradient might contribute to the sharp peaks obtained, high efficiency was mainly due to the use of poly( ethylene glycol) diacrylate as a biocompatible cross-linker.