Fabrication of a ferrocene-based monolithic column with a network structure and its application in separation of protein and small molecules

A novel ferrocene-based monolith with a network structure was fabricated via in situ free radical polymerization using vinyl ferrocene as the co-monomer within a stainless-steel column (50 × 4.6 mm i.d.) for the separation of proteins from complex bio-samples, taking merit of the specific absorption of ferrocene to protein, including human plasma, egg white, and standard proteins. The morphology and pore size distribution indicate that the optimized monolith has a relatively uniform structure with the network. The results showed that 26 fractions were separated from human plasma, and the column efficiency of the aromatic small molecule , naphthalene , was up to 30,560 plates m −1 . The homemade monolith showed excellent selectivity for intact proteins, mainly depending on the hydrophobic chromatography mechanism of ferrocene. In addition, the electrostatic interaction and hydrogen-bond interaction were the additional interactions in the chromatographic separation owing to the sandwich structure of ferrocene present in the monolithic column.