牛血清白蛋白亲和毛细管整体柱的制备及色氨酸对映体的分离

Preparation of Bovine Serum Albumin Affinity Monolith for Chiral Separation of D,L-tryptophan by Capillary Electrochromatography

利用亲和毛细管电色谱整体柱对色氨酸对映体进行了手性拆分研究.利用溶胶-凝胶法制备了壳聚糖/硅胶复合基质毛细管整体柱,然后采用戊二醛交联柱上键合牛血清白蛋白(BSA)得到亲和毛细管整体柱,以毛细管电色谱(CEC)模式分离色氨酸对映体,考察了缓冲液pH、浓度及有机修饰剂含量对分离过程的影响.在缓冲液的浓度为20 mmol/L,pH 7.5时,色氨酸对映体分离效果良好,分离度为2.44.

Enantioseparation of D,L-tryptophan was studied on an affinity monolithic column by capillary electrochromatography. A chitosan/silica hybrid monolithic column was prepared by the sol-gel method. Then the affinity stationary phase was synthesized by the in situ covalent immobilization of BSA on the monolithic column activated with glutaraldehyde. The monolith was used for chiral separation of D, L-tryptophan by capillary electrochromatography. The effects of p H, concentration of running buffer, and the content of organic modifier on the separation of D,L-tryptophan were investigated. The retention times of the solutes decrease and the resolution increases with the increase of buffer pH from 6, 0 to 8.0. With the increase of buffer concentration from 5 to 50 mmol/L,the retention times of the solutes increase,while the resolution increases to a higher value and then decreases. The organic modifier is not conducive to the chiral separation in this experimental system. A best separation of tryptophan enantiomers is achieved by using pH 7.5,20 mmol/L phosphate buffer,with the resolution of 2.44.