Fractionation separation of human plasma proteins using HPLC with a homemade iron porphyrin based monolithic column

In this work a polymer monolithic column was fabricated within the confines of a stainless steel column (50   ×   4.6   mm i.d.) via radical polymerization by using iron porphyrin and butyl methacrylate as co-monomers, ethylene glycol dimethacrylate as crosslinking agent, ethylene glycol, isopropyl alcohol and N, N-dimethylformamide as tri-porogens, benzoyl peroxide and N , N -dimethylaniline as initiators. The resulting monolithic column was characterized by elemental analysis , scanning electron microscopy, nitrogen adsorption BET surface area, and mercury intrusion porosimetry, respectively. Results showed that the homemade monolith occupied relatively uniform pore structure , low back pressure, and enhanced selectivity for proteins in complex bio-samples. The present work described a simple and efficient method for “fractionation separation” of human plasma proteins , and it is a promising separation method for complex bio-samples in proteomic research.