内嵌纳米粒阴离子交换聚甲基丙烯酸羟乙酯复合晶胶分离三磷酸胞苷

摘要

An anion exchange poly(2-hydroxyethyl methacrylate) composite cryogel embedded with SiO2 nanoparticles was prepared by cryo-polymerization followed by graft polymerization with monomer N,N-dimethylaminoethyl methacrylate. This cryogel was used to isolate cytidine triphosphate fromSaccharomyces cerevisiae transformation broth. Chromatographic separation was performed with 0.01 mol·L-1 HCl as running buffer together with 0.02 mol·L-1 and 0.1 mol·L-1 NaCl solutions (prepared in running buffer) as two-step elution liquids. Cytidine triphosphate purity was analyzed by high performance liquid chromatography. Cytidine triphosphate with high purity of 97.2% and recovery of 82.5% was obtained. Compared with other conventional methods, the present separation process was simple and effective, and could be used in separation of nucleotides from microbial fermentation or transformation feedstocks.%用结晶致孔法制备了内嵌SiO2颗粒的聚甲基丙烯酸羟乙酯基复合晶胶，经甲基丙烯酸二甲氨乙酯接枝修饰后，得到了阴离子交换复合晶胶介质，用于从啤酒酵母转化液中色谱分离三磷酸胞苷。色谱分离过程以0.01 mol·L-1 HCl为平衡液，以0.02 mol·L-1和0.1 mol·L-1的NaCl溶液（用平衡液配制）分步洗脱。通过高效液相色谱定量分析，所得三磷酸胞苷的纯度达97.2%，回收率82.5%。与其他分离方法相比，本法过程简单、分离迅速高效，在核苷酸药物的分离领域具有很好的应用前景。