【摘要】 报道了一种高效液相亲和色谱固定相的合成方法。以３－５μｍ的非多孔二氧化锆与脲醛树脂（ＵＴＲ）复合物微球用作高效液相亲和色谱固定相的基体，并用扫描电子显微镜检验该基体的结构和粒径。用此基体末端的酸胺基团为核心，分别与丙烯酸甲酯、乙二胺重复进行三次马氏加成反应和胺化反应，就形成末端为胺基的聚胺基酰胺（ＰＡＭＡＭ）星型树状间隔臂。用１３Ｃ固体核磁共振检测了树状间隔臂的拓扑结构。此间隔臂用丁二酸进行丁二酰化，并在１－乙基－３－（３－二甲基氨丙基）碳化二胺（ＥＤＡＣ）的存在下用Ｎ－羟基丁二酰亚胺（ＮＨＳ）活化，最后与配位体脱氧核糖核酸（ＤＮＡ）键合，并用紫外分光光度法测定了配位体容量。从而制备了一种［ＤＮＡ（ＰＡＭＡＭ）－（ＺｒＯ２－ＵＦＲ）］高效液相亲和色谱固定相。通过优化色谱柱操作参数，其已用于蛋白质的分析和核糖核酸（ＲＮＡ）碎片的分离。更多还原

【Abstract】 A method for the synthesis of high performance liquid affinity chromatographic (HPLAC) stationary phase was reported. The 3- 5 μm non-porous composite microspheres of zirconia and urea-formaldehyde (UF) resin were used as matrix of HPLAC stationary phase. The diameter and construction of the composite microspheres were judged by scanning electron microscope. Using the terminated amido-group of matrix as care, which was reacted respectively with methyl acrylate and ethylenediainine by repeating Michael’s addition reaction and amination three times, amine-terminated polyamidoamine (PAMAM) starburst dendrimer spacer arms were formed. The topological structure of the dendrimer spacer arms was inspected by solid-state 13C NMR. The spacer arms were succinylated by succinic acid, then activited by N-hydroxysuccinimide in the presence of 1 -ethyl-3-(3-dimethylaminopmpyl) carbodiimide (EDAC), and finally bonded deoxyribonucleic acid (DNA) ligand. The bonded amount of DNA was determined by UV spectrophotometry. A new type [DNA-( PAMAM ) -( Zirconia-UF resin)] stationary phase of I-IPLAC was prepared, which was used in the analysis of protein and the separation of ribonucleic acid (RNA) fragments by optimizing operation parameter of chromatographic column.更多还原