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.
