【摘要】 双回路技术是一套机械传动组件和流路系统,具有自动进样和馏分收集的双重功能。本研究基于双回路技术设计了一种二维液相色谱在线/离线接口,用以有效连接两种不同分离模式构建分离系统,提高系统峰容量,并对其功能进行了评价。以4种芳香族化合物为分离对象,利用设计的接口连接1套液相色谱系统,实现了进样、馏分收集、再进样纯度分析的多重操作集成;连接2套液相色谱系统,采用强阳离子交换模式对5种蛋白质样品进行初步分离,收集难分离组分,并通过再次进样到反相色谱模式的分离系统中,组分得到良好分离。进一步基于此接口连接1套强阳离子交换系统和1套微柱反相色谱系统,对牛血清白蛋白酶解产物进行在线二维分离,以1 m AU为积分阈值,共识别292个色谱峰。新型在线/离线接口可灵活实现样品微量制备、难分离组分的精细拆分和复杂样品的二维分离,为二维色谱系统的构建及分离研究提供了有力工具。更多还原

【Abstract】 By using a double loop technique based on a set of mechanical transmission components and a flow path system,a double function of injection and fraction collection was realized. On the basis of double loop technique,a novel type of online/offline interface for two-dimensional liquid chromatography was designed to construct an efficient separation system coupling two different separation modes with a higher peak capacity,and the functions of the interface were evaluated. By means of connecting the interface to an HPLC system,the multiple functions of injection,fraction collection and injection again were fulfilled for preparation and purity analysis of 4 kinds of aromatic compounds. As for combination of 2 sets of HPLC system,5 kinds of protein samples were separated preliminarily by strong cation exchange chromatography,and the components difficult to separate were collected and injected into reversed phase chromatographic system for further separation.Furthermore,the interface was applied to coupling two chromatographic systems in both strong cation exchange mode and microcolumn reversed-phase mode for the two-dimensional separation of bovine serum albumin enzymatic digest. When 1 m AU was set as the integral threshold,a total number of 292 peaks were identified.With the help of the online/offline interface,the preparation of microscale samples,fine separation of hardly separated samples and two-dimensional separation of complex samples were achieved flexibly. The result indicated that the system was a potent tool for the construction of two-dimensional chromatographic system and separation research.更多还原