Preparation of Titania Monoliths for HPLC by Titanium N-propoxide and Investigation of the Effect Factors on the Structure
|School||Tianjin University of Commerce|
|Course||Of Food Science|
|Keywords||Titania Monolithic column HPLC bovine serum albumin (BSA) trypsin polypeptide|
Titania has drawn increasing attention in recent years. Its unique properties have been extensively exploited in diverse areas, but the most striking property is its performance as a chromatographic packing material. Titania has great mechanical and chemical stability, it can be used in the separation of large quantities of samples, macromolecular compounds (such as polypeptide and protein) and the food safety testing, has a significant meaning. In this paper, titania monoliths were fabricated by sol-gel methods and the factors affect the structure were optimized. At the same time, the titania column was applyed in the separation of protein samples.The researches in the thesis included three aspects:(1) The titania monolithic material was prepared using sol-gel method by strictly controlling all steps in the experiments including the ratio of the initial mixtures, aging temperature, and the follow-up drying conditions. The effect factors on the structure and the performance of titania monolithic material were studied. The obtained monolithic material was characterized by surface area, pore size, SEM and XRD analyses.(2) By the single factor experiment of the bovine serum albumin (BSA) hydrolyzate obtained by trypsin, the hydrolyzate was separated by HPLC with a bar TiO2 column and the optimum conditions (the phosphate buffer concentrations of 15mM to firstly modify the bar TiO2 column, 85%ACN in eluents, the concentration is 3.0mM and pH=5.0) were obtained and the BSA hdrolyzate was separated accoding to the optimal conditions.(3) By the single factor experiment of bovine serum albumin (BSA) hydrolyzate obtained by trypsin, the hydrolyzate was separated by HPLC with a TiO2 RP-HPLC column and the optimum conditions (the concentrations of 40mM NH4F and higher ACN as the eluents) were obtained and it was also separated at the optimum conditions (buffer condition is 40mM NH4F and slow gradient ACN) by quaternary pump HPLC.