Synthesis Of Titania Monolithic Column And Its Application In Separation Of Protein And Peptide

Because of the unique properties of titania(TiO2), it has been extensively exploited in diverse areas such as catalysis, sensing, electronics, optics, and separation science. Recentle, researches on titania monolith have been further developed. Combining fine chemical properties of titania with high permeability of monoliths and applying to the stationary phase of high performance liquid chromatogram(HPLC), will cause the most interest in colour spectrum. Simultaneously, TiO2 as a new type of chromatographic stationary phase, there will be more application prospects of TiO2 in the biological materials, such as phospholipids, phosphoprotein, peptides, nucleic acid and intermediates as well as the analysis of carbohydrate. In this paper, synthesis of titania monolithic column and the optimization of conditions, as well as the application of titanium packing chromatographic column in the separation of phosphoprotein, peptides.The researches in this thesis included three aspects:(1) Using Ti(OnBu)4 as predecessor, acetic acid as chelating agent, n-butanol as solvent, hydrochloric acid as gel regulator, through sol-gel method, we synthesis of titania monolithic column. And we exosyndrome the material's performance we got. Accordingly, decide the factors that affect the microstructure of the material, including the ratio of water, n-butanol, hydrochloric acid and acetic acid,as well as their interaction. Gelatinization temperature and drying process are also be controled. At Ti/HAc/BuOH/HCl/H2O = 1/5/4/0.45/6 and 1/4/2.8/0.5/5.5, gelatinization temperature 21°C, we got titania monolithic column of better shape and performance.(2)Uesing NP(RP)-HPLC to separate zymohydrolysis products of casein. The best separating condition: NP-HPLC, ACN 85%, sodium phosphate buffer pH5.0, saltness2.0mM; RP-HPLC, ACN gradient: 3.5-15.4%(8min); 15.4-70%(32min); 70-35%(15min); 35%(5min).(3) Uesing NP(RP)-HPLC to separate macropeptides. The best separating condition: RP-HPLC, ACN gradient: 7-42%(20min); goto 70%(keep 5min),NH4F final concentration 80mM.