新型磷脂類有機基質(zhì)毛細管整體柱及應(yīng)用研究
發(fā)布時間：2019-06-06 17:15
【摘要】：仿生色譜法(Biomimetic Chromatography)廣義上講就是將生物活性分子通過合理設(shè)計成功能單體,然后修飾到色譜基質(zhì)上制備而成的色譜技術(shù),狹義上指的是固定化人工膜色譜法(Immobilized artificial chromatography,IAM),其中通過仿生色譜法制備的仿生磷脂單體制備的色譜柱,已經(jīng)被應(yīng)用于預(yù)測藥物-膜滲透性、蛋白質(zhì)純化和小分子化合物的分離,表現(xiàn)出優(yōu)異的色譜性能。整體柱被稱為“第四代色譜材料”,相對于傳統(tǒng)填充色譜柱,具有制備簡單、傳質(zhì)速度快、通透性好、分離效率高和易于改性等優(yōu)點。針對近年來越來越復(fù)雜的分析對象,基于有機基質(zhì)整體色譜柱制備簡單的特點,開發(fā)豐富多樣的磷脂仿生整體色譜柱,結(jié)合和發(fā)揮整體色譜柱與磷脂固定相的優(yōu)點是研究的熱點方向。目前基于磷脂單體仿生色譜柱主要是硅膠色譜柱,而且種類稀少,藥物-膜相互作用預(yù)測能力差。本文基于課題組前期研究工作基礎(chǔ),針對性地設(shè)計合成了兩種磷脂修飾功能單體,并將其成功制備了不同功能化的新型有機基質(zhì)毛細管整體柱,并進行了基礎(chǔ)應(yīng)用研究。第一章,首先介紹仿生色譜法的理論及其應(yīng)用,提出基于磷脂制備仿生色譜柱的意義與待解決的問題;接著介紹整體柱的發(fā)展歷史和應(yīng)用開發(fā),根據(jù)整體柱基質(zhì)的不同,分為無機硅膠整體柱、有機聚合物整體柱和有機無機雜化整體柱三大類進行講述,并重點對有機聚合物混合作用模式和親水作用整體柱做了詳細的介紹;最后介紹新型聚合體系的有機聚合物整體柱——單一交聯(lián)劑功能單體制備的高交聯(lián)整體柱及其發(fā)展與應(yīng)用。第二章,通過對細胞膜酸性磷脂——磷脂酸PA進行半合成結(jié)構(gòu)修飾,得到C_(12)的單鏈磷脂酸:2-甲基丙烯酰氧十二烷基磷脂酸(12-Methacryloyl dodecylphosphatidic acid,MDPA),并作為為功能化單體制備得到poly(MDPA-co-EDMA)整體柱。首先通過半經(jīng)驗法優(yōu)化制備條件,得到綜合色譜性能最佳的整體柱;然后考察整體柱的理化性質(zhì)和色譜性能;接著對整體柱的色譜相互作用機制進行深入研究,整體柱具有陽離子交換/反相混合作用雙重作用,相比單一陽離子交換或者反相作用模式色譜柱,混合模式色譜柱結(jié)合兩種色譜作用力,具有更好的選擇性和分離能力;最后選取不同理化性質(zhì)的標(biāo)準(zhǔn)分析物對整體柱進行分離能力的考察。第三章,通過對細胞膜中主要成分——磷脂酰膽堿PC進行半合成結(jié)構(gòu)修飾,得到C2的雙交聯(lián)磷脂酰膽堿:N-(2-甲基丙烯酰氧)-2-甲基丙烯酰氧磷酸膽堿(2-(methacryloyloxy)ethyl-[N-(2-methacryloyloxy)ethyl]phosphorylcholine,MMPC),并以MMPC為功能單體和交聯(lián)劑制備得到poly MMPC整體柱。首先考察聚合時間對整體柱制備的影響,聯(lián)系分析了比表面積、孔徑分布曲線和孔隙率與色譜性能的關(guān)系,并得出1 h的快速制備時間;接著通過半經(jīng)驗法優(yōu)化聚合溶液配比,得到綜合色譜性能最佳的整體柱;然后考察整體柱的理論性質(zhì)和色譜性能;然后對整體柱的色譜相互作用機制進行深入研究,結(jié)果顯示整體固定相表現(xiàn)出強親水作用及微弱靜電作用,ACN:H2O(20~95%,v/v)的流動相,色譜柱都呈現(xiàn)親水作用;最后選取不同理化性質(zhì)的標(biāo)準(zhǔn)分析物對整體柱進行分離能力的考察。第四章,總結(jié)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.
【學(xué)位授予單位】：廣東藥科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O652.63
本文編號：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.
【學(xué)位授予單位】：廣東藥科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O652.63
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