Preparation Of Hybrid Silica Monolithic Column Modified By Bovine Serum Albumin And Application In Separation Of Chiral Compounds

A novel hybrid monolithic column(HMC)was prepared in a conventional stainless steel column with using methyltrimethoxysilane as single silane,methanol as solvent and nitric acid as catalyst.The HMC has a stable network structure and a controllable size.The HMCs of different pore sizes and framework diameters were prepared by optimizing the conditions.After being modified,the columns were covalently bonded with bovine serum albumin and three chiral monolithic columns were successfully prepared.The chiral monolithic columns were coupled with liquid chromatography for resolution of chiral compound enantiomers,which successfully achieved baseline separation of chiral molecules and chiral drug molecules by optimization of chromatographic conditions.Chapter 1: The meaning of chiral separation,the methods of resolution of chiral compounds and the synthesis of chiral stationary phases,as well as the research progress of the chiral monolithic column were described.Chapter 2: A novel HMC was prepared by sol-gel method using methyltrimethoxysilane as silicon source,methanol as solvent and nitric acid as catalyst.The conditions for preparing the monolithic column were optimized.The 3-aminopropyltrimethoxysilane is grafted into the HMC to prepare an aminated column.Then the columns were reacted with glutaraldehyde to prepare the columns having an aldehyde group on the surface.Finally,bovine serum albumin is dynamically grafted onto the columns.The columns were characterized by scanning electron microscopy and infrared spectroscopy.The results showed that the monolithic column obtained controlled pores and the target materials were successfully grafted.The conditions for dynamic grafting method were investigated,and the maximum grafting amount was 48.6 mg/g.The HMCs were coupled with high performance liquid chromatography.Under the optimized chromatographic conditions,the enantiomeric separation of the chiral substance tryptophan was achieved successfully.The resolution was 3.05.The HMCs also successfully achieved baseline separation of the chiral drug Pidotimod.Chapter 3: A novel chiral monolithic column was prepared using a monolithic column as matrix based on the ring opening reaction of epoxy groups.The stationary phase was combined with liquid chromatography and successfully applied to the separation of chiral substances.The HMC was prepared in conventional stainless steel columns.The ?-(2.3-epoxypropyl)propyltrimethoxysilane was grafted onto the surface of HMC.Then,the aldehyde group was formed on the surface of the monolith by oxidation reaction of periodic acid.Finally,the bovine serum albumin was immobilized on the hybrid monolith by Schiff base.The columns were characterized by scanning electron microscopy and infrared spectroscopy.The results showed that the column structure is stable and the target materials were successfully grafted.The conditions for grafting bovine serum albumin were optimized.Its maximum grafting amount is 58.3 mg/g.The columns were coupled with with liquid chromatography.Under optimized chromatographic conditions,the baseline separation of the chiral substance tryptophan was achieved with a maximum resolution of 4.78.The baseline separation of the chiral drug molecule atenolol was also achieved successfully.Chapter 4: The HMC with aldehyde groups on the surface is reacted with polyethyleneimine to have a large number of amino groups on the surface.The columns were reacted with glutaraldehyde,and the bovine serum albumin is grafted onto the modified hybrid monolith by Schiff base.The prepared HMC was characterized by scanning electron microscopy and infrared spectroscopy.The results showed that the column structure was stable and the target materials were successfully grafted.The grafting conditions were optimized with a maximum grafting amount of 56.1 mg/g.The HMCs were coupled with liquid chromatography.Under optimized chromatographic conditions,the baseline separation of the chiral substance benzoin was achieved with a maximum resolution of 3.78.The chiral stationary phase shows better method repeatability and reusability.