Preparation And Chromatographic Characteristics Of HPLC Stationary Phase For Bioseparation

Bioseparation process is dominated by chromatographic steps. HPLC stationary phase is very useful in HPLC separation. So it is important to study HPLC stationary phase. In my research I mainly studied Macro-porous silica gel stationary phase as separating media for separation of proteins. Macro-porous silica gel is reacted with silanes for column packing. Two types of stationary phases are prepared; 1) Silica-bonded stationary phase; 2) Silica-based-polymer bonded stationary phase. Chromatographic characteristics of stationary phases are also studied.1. Macro-porous silica gel reacted with silanes KH560 and silica bonded stationary phase is prepared. It is silica-bonded silane-KH560 stationary phase. Then trimethylchlorosilane (Si(CH₃)₃ Cl) reacted with residual Si-OH to make it inactive. Epoxy compounds in KH560 bonded silica gel stationary phase are hydrolyzed. Column is packed and applied to separation of proteins. It shows good results. This stationary phase is inactive with proteins and does not destroy the nature of proteins. So it is suitable for separation of proteins. This column has also good permeability. It can be used for quick and high efficient separation of proteins.2. Two types of silica-bonded stationary phases having Alkenes are prepared. One is KH570 bonded silica gel. Another is vinyl bonded silica gel. These types of silica gel are reacted with 1,4-divinylbenzene and 2-Hydroxypropyl methacrylate to make silica-based-polymer-bonded phase. Optimum reaction conditions are selected. Then column was packed and applied to separation of proteins. Polymerization has no effect on macro-porous. This column has good permeability. Silica-based-polymer-bonded stationary phase is also inactive with proteins and does not destroy the nature of protein. It can be used for quick and high efficient separation of proteins. This stationary phase is also stable in basic solvent.