新型磷脂类有机基质毛细管整体柱及应用研究
发布时间：2019-06-06 17:15
【摘要】：仿生色谱法(Biomimetic Chromatography)广义上讲就是将生物活性分子通过合理设计成功能单体,然后修饰到色谱基质上制备而成的色谱技术,狭义上指的是固定化人工膜色谱法(Immobilized artificial chromatography,IAM),其中通过仿生色谱法制备的仿生磷脂单体制备的色谱柱,已经被应用于预测药物-膜渗透性、蛋白质纯化和小分子化合物的分离,表现出优异的色谱性能。整体柱被称为“第四代色谱材料”,相对于传统填充色谱柱,具有制备简单、传质速度快、通透性好、分离效率高和易于改性等优点。针对近年来越来越复杂的分析对象,基于有机基质整体色谱柱制备简单的特点,开发丰富多样的磷脂仿生整体色谱柱,结合和发挥整体色谱柱与磷脂固定相的优点是研究的热点方向。目前基于磷脂单体仿生色谱柱主要是硅胶色谱柱,而且种类稀少,药物-膜相互作用预测能力差。本文基于课题组前期研究工作基础,针对性地设计合成了两种磷脂修饰功能单体,并将其成功制备了不同功能化的新型有机基质毛细管整体柱,并进行了基础应用研究。第一章,首先介绍仿生色谱法的理论及其应用,提出基于磷脂制备仿生色谱柱的意义与待解决的问题;接着介绍整体柱的发展历史和应用开发,根据整体柱基质的不同,分为无机硅胶整体柱、有机聚合物整体柱和有机无机杂化整体柱三大类进行讲述,并重点对有机聚合物混合作用模式和亲水作用整体柱做了详细的介绍;最后介绍新型聚合体系的有机聚合物整体柱——单一交联剂功能单体制备的高交联整体柱及其发展与应用。第二章,通过对细胞膜酸性磷脂——磷脂酸PA进行半合成结构修饰,得到C_(12)的单链磷脂酸:2-甲基丙烯酰氧十二烷基磷脂酸(12-Methacryloyl dodecylphosphatidic acid,MDPA),并作为为功能化单体制备得到poly(MDPA-co-EDMA)整体柱。首先通过半经验法优化制备条件,得到综合色谱性能最佳的整体柱;然后考察整体柱的理化性质和色谱性能;接着对整体柱的色谱相互作用机制进行深入研究,整体柱具有阳离子交换/反相混合作用双重作用,相比单一阳离子交换或者反相作用模式色谱柱,混合模式色谱柱结合两种色谱作用力,具有更好的选择性和分离能力;最后选取不同理化性质的标准分析物对整体柱进行分离能力的考察。第三章,通过对细胞膜中主要成分——磷脂酰胆碱PC进行半合成结构修饰,得到C2的双交联磷脂酰胆碱:N-(2-甲基丙烯酰氧)-2-甲基丙烯酰氧磷酸胆碱(2-(methacryloyloxy)ethyl-[N-(2-methacryloyloxy)ethyl]phosphorylcholine,MMPC),并以MMPC为功能单体和交联剂制备得到poly MMPC整体柱。首先考察聚合时间对整体柱制备的影响,联系分析了比表面积、孔径分布曲线和孔隙率与色谱性能的关系,并得出1 h的快速制备时间;接着通过半经验法优化聚合溶液配比,得到综合色谱性能最佳的整体柱;然后考察整体柱的理论性质和色谱性能;然后对整体柱的色谱相互作用机制进行深入研究,结果显示整体固定相表现出强亲水作用及微弱静电作用,ACN:H2O(20~95%,v/v)的流动相,色谱柱都呈现亲水作用;最后选取不同理化性质的标准分析物对整体柱进行分离能力的考察。第四章,总结和展望。
[Abstract]:In a broad sense, that biomimetic chromatography is a chromatographic technique, in a narrow sense, of an immobilized artificial membrane chromatography (iam), which is prepared by a reasonable design of a successful monomer and then to a chromatographic matrix. The chromatographic column prepared by the bionic phospholipid monomer prepared by the bionic chromatography has been applied to the separation of the predicted drug-membrane permeability, the protein purification and the small molecular compound, and exhibits excellent chromatographic performance. The monolithic column is called a "fourth-generation chromatography material", and has the advantages of simple preparation, high mass transfer speed, good permeability, high separation efficiency and easy modification, and the like relative to the traditional packed chromatographic column. Aiming at the more and more complex analysis objects in recent years, based on the simple characteristics of the whole column of the organic matrix, the development of the rich and varied phospholipid biomimetic monolithic column, the combination and the advantages of the whole chromatographic column and the fixed phase of the phospholipid is the hot spot direction of the research. Currently, the bionic chromatographic column based on the phospholipid monomer is mainly a silica gel column, and the type is small, and the drug-membrane interaction prediction is poor. In this paper, based on the previous research work of the research group, two kinds of phospholipid-modified functional monomers were designed and synthesized, and the new organic matrix capillary whole column with different function was successfully prepared and the basic application was studied. In the first chapter, the theory and application of the biomimetic chromatography are introduced, and the significance of the preparation of the bionic chromatographic column based on the phospholipid and the problems to be solved are put forward. The development history and the application development of the whole column are introduced, and the whole column of the inorganic silica gel is divided into the whole column of the inorganic silica gel according to the difference of the whole column matrix. The organic polymer monolithic column and the organic and inorganic hybrid monolithic column are described, and the mixed mode of organic polymer and the whole column of hydrophilic action are introduced in detail. In this paper, the high-crosslinking monolithic column prepared by the single-cross-linking agent functional monomer of the organic polymer of the new type of polymerization system and its development and application are introduced. In the second chapter, by the semi-synthetic structure modification of the cell membrane acid-phospholipid-phosphatidic acid PA, the single-chain phosphatidic acid of C _ (12) is obtained:2-methylpropyleneoxy-12-alkyl phosphatidic acid (MDPA), and the poly (MDPA-co-EDMA) monolithic column is prepared as a functional monomer. firstly, the preparation conditions are optimized through a semi-empirical method to obtain an overall column with the best comprehensive chromatographic performance; then, the physical and chemical properties and the chromatographic performance of the whole column are examined; and then the chromatographic interaction mechanism of the whole column is further researched, The monolithic column has a double effect of cation exchange/ reverse-phase mixing, and compared with a single cation exchange or a reversed-phase action mode chromatographic column, the mixed mode chromatographic column is combined with two chromatographic forces, and has better selectivity and separation capability; And finally, the standard analyte of different physical and chemical properties is selected to carry out the investigation on the separation capability of the whole column. In the third chapter, by the semi-synthetic structure modification of the main components of the membrane in the cell membrane, the double-cross-linked phospholipid-choline: N-(2-methylpropyleneoxy) -2-methylpropyleneoxy-phosphate (2-methylpropyloxy) ethyl-[N-(2-methacrylxy) ethyl] phosphate (MMPC) was obtained. And a poly (MMPC) monolithic column is prepared by using the MMPC as a functional monomer and a cross-linking agent. Firstly, the effect of the polymerization time on the preparation of the monolithic column was investigated, the relationship between specific surface area, pore size distribution curve and porosity and the chromatographic performance was analyzed, and the rapid preparation time of 1 h was obtained. Then, the ratio of the polymerization solution was optimized by the semi-empirical method. and then the theoretical property and the chromatographic performance of the whole column are examined, the chromatographic interaction mechanism of the whole column is further researched, the result shows that the whole fixed phase exhibits strong hydrophilic action and weak electrostatic effect, and the ACN: H2O (20-95%, V/ v) the mobile phase and the chromatographic column show the hydrophilic action; and finally, the standard analyte of different physical and chemical properties is selected to carry out the investigation on the separation capability of the whole column. Chapter 4, Summary and Prospect.
【学位授予单位】：广东药科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O652.63
本文编号：2494465
[Abstract]:In a broad sense, that biomimetic chromatography is a chromatographic technique, in a narrow sense, of an immobilized artificial membrane chromatography (iam), which is prepared by a reasonable design of a successful monomer and then to a chromatographic matrix. The chromatographic column prepared by the bionic phospholipid monomer prepared by the bionic chromatography has been applied to the separation of the predicted drug-membrane permeability, the protein purification and the small molecular compound, and exhibits excellent chromatographic performance. The monolithic column is called a "fourth-generation chromatography material", and has the advantages of simple preparation, high mass transfer speed, good permeability, high separation efficiency and easy modification, and the like relative to the traditional packed chromatographic column. Aiming at the more and more complex analysis objects in recent years, based on the simple characteristics of the whole column of the organic matrix, the development of the rich and varied phospholipid biomimetic monolithic column, the combination and the advantages of the whole chromatographic column and the fixed phase of the phospholipid is the hot spot direction of the research. Currently, the bionic chromatographic column based on the phospholipid monomer is mainly a silica gel column, and the type is small, and the drug-membrane interaction prediction is poor. In this paper, based on the previous research work of the research group, two kinds of phospholipid-modified functional monomers were designed and synthesized, and the new organic matrix capillary whole column with different function was successfully prepared and the basic application was studied. In the first chapter, the theory and application of the biomimetic chromatography are introduced, and the significance of the preparation of the bionic chromatographic column based on the phospholipid and the problems to be solved are put forward. The development history and the application development of the whole column are introduced, and the whole column of the inorganic silica gel is divided into the whole column of the inorganic silica gel according to the difference of the whole column matrix. The organic polymer monolithic column and the organic and inorganic hybrid monolithic column are described, and the mixed mode of organic polymer and the whole column of hydrophilic action are introduced in detail. In this paper, the high-crosslinking monolithic column prepared by the single-cross-linking agent functional monomer of the organic polymer of the new type of polymerization system and its development and application are introduced. In the second chapter, by the semi-synthetic structure modification of the cell membrane acid-phospholipid-phosphatidic acid PA, the single-chain phosphatidic acid of C _ (12) is obtained:2-methylpropyleneoxy-12-alkyl phosphatidic acid (MDPA), and the poly (MDPA-co-EDMA) monolithic column is prepared as a functional monomer. firstly, the preparation conditions are optimized through a semi-empirical method to obtain an overall column with the best comprehensive chromatographic performance; then, the physical and chemical properties and the chromatographic performance of the whole column are examined; and then the chromatographic interaction mechanism of the whole column is further researched, The monolithic column has a double effect of cation exchange/ reverse-phase mixing, and compared with a single cation exchange or a reversed-phase action mode chromatographic column, the mixed mode chromatographic column is combined with two chromatographic forces, and has better selectivity and separation capability; And finally, the standard analyte of different physical and chemical properties is selected to carry out the investigation on the separation capability of the whole column. In the third chapter, by the semi-synthetic structure modification of the main components of the membrane in the cell membrane, the double-cross-linked phospholipid-choline: N-(2-methylpropyleneoxy) -2-methylpropyleneoxy-phosphate (2-methylpropyloxy) ethyl-[N-(2-methacrylxy) ethyl] phosphate (MMPC) was obtained. And a poly (MMPC) monolithic column is prepared by using the MMPC as a functional monomer and a cross-linking agent. Firstly, the effect of the polymerization time on the preparation of the monolithic column was investigated, the relationship between specific surface area, pore size distribution curve and porosity and the chromatographic performance was analyzed, and the rapid preparation time of 1 h was obtained. Then, the ratio of the polymerization solution was optimized by the semi-empirical method. and then the theoretical property and the chromatographic performance of the whole column are examined, the chromatographic interaction mechanism of the whole column is further researched, the result shows that the whole fixed phase exhibits strong hydrophilic action and weak electrostatic effect, and the ACN: H2O (20-95%, V/ v) the mobile phase and the chromatographic column show the hydrophilic action; and finally, the standard analyte of different physical and chemical properties is selected to carry out the investigation on the separation capability of the whole column. Chapter 4, Summary and Prospect.
【学位授予单位】：广东药科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O652.63
【参考文献】
相关期刊论文 前6条
1 戴荣继;王慧婷;孙维维;邓玉林;吕芳;刘秀洁;;高分子手性固定相的研究进展[J];色谱;2016年01期
2 冯颖淑;童珊珊;徐希明;余江南;;亲和色谱应用于天然活性成分筛选的研究进展[J];中国中药杂志;2015年06期
3 窦阿波;邹存杰;单圆鸿;石先哲;许国旺;;混合模式毛细管聚合物整体柱的制备及应用[J];色谱;2014年05期
4 张振宾;欧俊杰;林辉;刘忠山;董靖;邹汉法;;有机-硅胶杂化整体柱的制备及应用研究进展[J];高等学校化学学报;2013年09期
5 杨更亮;刘海燕;;高效液相色谱整体柱在药物分离分析中的应用进展[J];中国科学:化学;2010年06期
6 邓玉林,刘艳清,马文,历红;生物功能化色谱新概念及其方法学研究[J];生命科学仪器;2004年05期
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