Preparation and Utilization of Monolithic Column as HPLC Stationary Phase for Alkyl Benzene Separation with Low Mobile Phase Usage

摘要

One of the long-known separation methods is chromatography, such as high performance liquid chromatography (HPLC). This separation essentially uses column containing particle-packed which generates high flow-resistance and low mass transfer. As matter of fact, it has an impact to high amount mobile phase usage. Meanwhile, organic polymer monolithic column has been widely used as alternative due to its high mass transfer. In addition, the resulting column has stability over wide pH range, high temperature, shrinkage and swelling of the reservoir. The aim of this research was to produce monolithic column which able to utilized for separation with low amount of mobile phase usage. Preparation of monolithic column begins with pretreatment of polyetereterketone tubing column inner wall. It conducted by activating the inner wall with H_2SO_4 49% (v/v), followed by vinylization with glycidyl methacrylate. Furthermore, pretreated column filled by polymer mixture consisted of 30 wt% of monomers glycidyl methacrylate:trimetilolpropane trimethacrylate 4:1 (w/v), 70 wt% of pore-forming agents (l-propanol/1,4-butanediol/water 7:4:1 w/v), and a,a'-azoisobutyronitrile 1 wt% of total monomers amount. The polymerization conducted at 60 °C for 12 h. Produced column connected to HPLC then applied to separate toluene and amylbenzene. The result shows both compounds successfully separated by 7.5 μL/min flow rate of acetonitrile:water (75:25) at 84 and 132 min respectively.