奎宁衍生物键合杂化介孔硅胶手性固定相的制备及应用研究

发布时间：2018-06-15 20:26

本文选题：高效液相色谱法 + 奎宁衍生物　；参考：《郑州大学》2014年硕士论文

【摘要】：手性是自然界的物质属性之一。大多数手性药物以外消旋体的形式存在。而单一构型的药物由于其作用位点明确，见效快，已经成为药物研发的璀璨明星。因而，各种新型的手性药物的分离与分析技术应运而生。其中，手性固定相法应用最为广泛。球形氨丙基乙基桥连介孔硅胶（APEPMOs）中不仅含有氨丙基和乙撑基两种官能团，球形形貌，而且具有高度有序的介孔通道、较大的比表面积、较高的容量和机械强度，不仅可以直接作为常规液相色谱柱填料，并且其侧链上的氨丙基为富电子的活性官能团，可以为后续的衍生化修饰提供便利；其通透性较好，在快速分离分析方面有一定的优势。而目前为止，成功制备球形氨丙基修饰的PMOs的报道还比较少。而对于手性选择剂奎宁而言，一直以来，人们都致力于制备奎宁、奎宁衍生物键合手性固定相及正相条件下拆分手性对映体的研究，但很少有文献报道在反相洗脱模式下，考察流动相变量（如流动相组成等）对手性化合物拆分的影响。因此，手性选择剂奎宁衍生物与杂化介孔硅胶的结合，和对其在反相条件下应用的研究有着理论基础和必要性。本文旨在开发一种新型手性固定相，为建立高效、快速的手性药物拆分方法提供新型的分离材料。以APEPMOs为固定相的键合基质，以QN为手性选择剂，将QN键合到介孔硅胶上，制备出新型奎宁衍生物键合手性固定相，并将其制备成手性色谱柱，评价了自制手性柱的色谱性能及在反相洗脱模式下对多种手性药物的拆分能力。首先，以1,2-二（三乙氧基硅基）乙烷（BTSE）和3-氨丙基三乙氧基硅烷（KH-550）为双硅源，以阳离子表面活性剂十八烷基三甲基氯化铵（C18TACl）为模板，，乙醇为助溶剂，在碱性条件下通过一步共缩聚合成新型氨丙基乙基桥连球形介孔硅胶；再以偶氮二异丁腈为引发剂，与O-9-（叔丁酯氨基甲酰）奎宁（QN）反应，合成奎宁衍生物键合新型杂化介孔硅胶手性固定相。并对每一步的反应产物进行相应的纯化和表征，表征方法主要有傅里叶红外分光光度计（FT-IR）、元素分析、扫描电子显微镜（Scanning electron microscope, SEM）以及氮气吸附实验，最终得到结构明确的手性固定相。采用湿法装柱，以三氯甲烷-二氧六环（50/50, V/V）为匀浆液，将合成的手性固定相填装于不锈钢液相色谱柱中，制得新型手性液相色谱柱。其次，研究自制奎宁衍生物键合氨丙基介孔硅胶手性色谱柱在高效液相色谱（HPLC）中的应用。在反相洗脱模式下，扁桃酸及其衍生物邻氯扁桃酸、酮洛芬3种手性药物达到了基线分离（Rs1.5），并系统地考察了流动相改性剂的浓度、柱温及流速等因素对手性药物对映体分离的影响。最后，本文考察了自制手性色谱柱的重复性和稳定性，对三个不同批次的自制手性色谱柱进行考察，得知其手性识别能力及柱效基本保持一致，从而说明自制手性柱的重复性较好；通过对同一自制手性色谱柱在不同时间段色谱性能的考察，发现其手性识别能力减弱，柱效有所降低，说明其稳定性不太好。可能由于键合量问题，部分药物对映体未能达到基线分离。
[Abstract]:Chiral is one of the material properties of nature. Most of the chiral drugs exist in the form of racemes. And single configuration drugs have become the resplendent star of drug research and development because of their clear action sites and fast effect. Therefore, the separation and analysis techniques of various new chiral drugs have emerged. For a wide range.
Spherical amino propyl ethyl bridged mesoporous silica gel (APEPMOs) not only contains two kinds of functional groups, such as propyl and ethopyl groups, spherical morphology, highly ordered mesoporous channels, larger specific surface area, higher capacity and mechanical strength, not only can be directly used as conventional liquid chromatography column filler, but also its side chain is rich in propyl. The active functional groups of the children can provide convenience for subsequent derivatization, and their permeability is good and has some advantages in rapid separation and analysis. So far, there are few reports on the successful preparation of spherical modified PMOs. The study of chiral chiral stationary phase and chiral enantiomer under normal phase is studied by Ningyan, but few reports have been reported on the effect of the separation of chiral compounds in the reverse phase elution mode, such as the composition of mobile phase, etc. There is a theoretical basis and necessity for the research under the application.
The aim of this paper is to develop a new chiral stationary phase, which provides a new separation material for the establishment of high efficient and fast chiral separation method. Using APEPMOs as the bonding matrix and QN as the chiral selector, the QN bond is bonded to the mesoporous silica gel to prepare a new type of chiral stationary phase of quinine derivatives, and the chiral chromatography is prepared. The chromatographic properties of the self-made chiral column and the resolution ability of chiral drugs in the reverse elution mode were evaluated.
First, 1,2- two (triethoxyl silicyl) ethane (BTSE) and 3- ammonia propyl triethoxy silane (KH-550) were used as two silicon sources, with cationic surfactant eighteen alkyl three methyl ammonium chloride (C18TACl) as the template and ethanol as a cosolvent, a new type of propyl ethyl bridge spherical mesoporous silica gel was synthesized by one step co polycondensation under alkaline conditions. Nitrogen two isobutyl nitrile was used as an initiator to react with O-9- (tert butyl amino formyl) quinine (QN) to synthesize quinine derivatives bonding new hybrid mesoporous silica chiral stationary phase. The reaction products of each step were purified and characterized. The main characterization methods were Fourier infrared spectrophotometer (FT-IR), element analysis, scanning electron microscope (Sc). Anning electron microscope, SEM) and nitrogen adsorption experiments were carried out. Finally, the chiral stationary phase with clear structure was obtained. The chiral stationary phase was filled in the stainless steel liquid chromatography column using the wet packed column and the trichloromethane - two oxygen six ring (50/50, V/V) as the homogenate.
Secondly, the application of the homemade quinine derivative bonded mesoporous silica chiral chromatographic column in high performance liquid chromatography (HPLC) was studied. Under the reverse phase elution mode, 3 kinds of chiral drugs, almanic acid and its derivatives, oalmanic acid and ketoprofen, reached the baseline separation (Rs1.5), and the concentration, column temperature and flow velocity of the mobile phase modifier were systematically investigated. The effect of adversary drug on enantiomer separation.
Finally, the reproducibility and stability of the homemade chiral chromatographic column were investigated, and the self-made chiral chromatographic columns of three different batches were investigated. It was found that the chiral recognition ability and the column efficiency were basically consistent, which indicated that the reproducibility of the homemade chiral column was better, and the performance of the chromatographic column of the same self made chiral chromatographic column was in different time periods. It is found that the chiral recognition ability is weakened and the column efficiency is reduced, indicating that its stability is not very good.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R943

【参考文献】