A Chromatographic Separation System Based On Diazo Resin And Vancomycin

Our lives are inseparable from chirality and chiral analysis is also one of the topics of great concern in various fields.Chiral stationary phases characterized by their specificity and stability are often the first choice of people during the separation process.Among them,antibiotic functionalized stationary phases have been successfully applied to the separation of enantiomers of various chiral compounds.Considering that liquid chromatography and capillary electrophoresis are the most commonly used methods for chiral separation,herein,photosensitive diazoresin(DR)is used as a coupling agent and the amphiphilic glycopeptide molecule vancomycin is modified on the surface of both porous polymer microspheres and the inner wall of the capillary.Thus,two different separation systems of modified high-performance liquid chromatography stationary phase and coated capillary electrophoresis were constructed,and the separation performance of different separation systems was explored respectively.The work carried out in this paper includes:1.The synthetic route of porous microspheres is designed to prepare diazoresin / vancomycin modified polymer porous microspheres with a particle size of 4.5 μm.The effect of different hydrolysis conditions on the performance of polymer porous microspheres during hydrolysis modification process was studied which is of great guiding significance for the subsequent modification design on the surface of porous microsphere.2.Diazo resin / vancomycin-modified polymer porous microspheres are used as highperformance liquid chromatography stationary phases.The relationship between the column pressure and flow rate of the column was explored and the chiral separation of four chiral drugs was performed.In addition,chromatographic separation conditions were optimized and the column pressure was relatively low.Finally,the baseline separation of chlorpheniramine,benzoin and promethazine were successfully achieved under the mobile phase of acetonitrile/triethylamine buffer.This work broadens the scope of application of antibiotic functional chiral columns.3.The internal wall of the capillary was modified with vancomycin to form a new capillary coating.The biggest advantage of the coated capillary is that protein separation and chiral separation can be achieved simultaneously.It not only realizes the separation of four proteins but also the baseline separation of several chiral substances,which solves problems of low separation column efficiency,high cost and high sample loss in the separation of chiral substances.