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。
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.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2001年第10期1129-1134,共6页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金资助课题(No.29775005)
