Study On Synthesis,Characteristies And Applications Of A New HIC Packing Material——Butyl Cross Linked Chitosan

Hydrophobic Interaction Chromatography (HIC) is a new Chromatographic technique developed in the 1970's to separate biopolymers. Generally, Sepharose is used as the carrier, and using chitosan as a carrier has not been reported. In this paper, a new packing material of HIC-Butyl-Cross linked Chitosan (CTS CL-C4) has been synthesized which was prepared with Chitosan as the carrier, butyl as the hydrophobic ligand and gluteraldehyde as the cross-linker to raise the mechanical intensity. The control of the synthetic conditions has been researched and series of characteristics and application of CTS CL-C4 have been determined. The results are shown as the follows: 1. The concentration of gluteraldehyde, reactant proportion and reactant consistency determinate the hydrophobic nature of the packing material. For good operation and practical nature of guaranty, the more suitable synthetic conditions are selected. The last concentration of gluteraldehyde is 0.1 --0.15% (V/V); the concentration of Chitosan is 0.2-1.0%(W/V), that is 2-10g/L; the concentration of butyraldehyde is 0.13%-0.26%(V/V); mol-proportion of aldehyde and aniido is O.5:l-1:l; synthetic temperature is in 40℃-70℃; stirring intermittently. Concrete protein is defined the corresponding synthetic conditions on the basis of its hydrophobic nature. 2. Characteristics of CTS CL-C4 have been determined using bovine serum albumin (BSA) and C?-Amylase as HIC target: the adsorption of BSA increases with the increasing of salt concentration, (NH4)2SO4 is often used to get better effect; selection of adsorption time and temperature is 0.5-1hr and 20℃-25℃; effect of pH is complex and varied and under more research. 3. Crude ci -Amylase from Bacillus Subtilis can be effectively purified on this packing material with the enzyme activity recovered in 72.62% yield, protein recovered in 94.67% yield and a 1.7-fold purification.