【作者】 刘鹏；

【导师】 张生勇；

【作者基本信息】 第四军医大学 ， 药理学， 2008， 博士

【摘要】 高效毛细管电泳（HPCE）是近年来发展起来的一种简便、高效、灵敏、快速和微量的液相微分离分析技术,广泛应用于生命科学、化学和药学等研究领域,特别是在手性化合物分离、药物分析、生物大分子分析和环境分析等领域受到人们越来越多的关注,现已成为手性药物分离中有效的检测手段之一。在毛细管电泳手性分析中,通常是将手性选择剂加入背景电解质中构建手性分离环境,在分离过程中由于不同构型的对映体与手性选择剂的相互作用不同,导致不同对映体的有效电泳淌度不同,最终达到分离的目的。本论文以天然β-环糊精（β-CD）为基础,合成了4种新型手性选择剂,并用于毛细管电泳分离和测定手性邻二醇、手性芳香仲醇和手性氨基酸等多种手性化合物,并对手性分离的原理进行了初步探讨。主要研究工作有以下几个方面:1.在文献基础上,以CaH₂为缚酸剂,合成了用于不对称双羟基化反应（AD反应）的手性配体（QN）2PHAL,将其用于8种潜手性烯烃的AD反应,取得了良好的实验结果。在此基础上,以天然β-CD作为毛细管电泳手性选择剂,建立了快速有效的分离8种烯烃AD反应产物对映体的新方法,并探讨了影响手性分离的若干原因。在优化的电泳条件下,所有邻二醇对映体在15min内都实现了基线分离。用所建立的方法测定了AD反应合成样品的化学纯度和百分对映体过量值（%ee）。该法的测定结果与HPLC测定结果相一致。2.通过对β-环糊精结构中C6-位的修饰,合成了2种阳离子型氨基化-β-环糊精（β-CDen和β-CDdien）手性选择剂。对氨基化-β-环糊精合成过程中的重要中间体（6-OTs-β-CD）的合成方法进行了改进,简化了操作步骤,将化学产率从11%提高到23%。然后在DMAP催化作用下该中间体分别与过量的乙二胺和二亚乙基三胺作用,分别以50.3%和51.6%的产率得到β-CDen和β-CDdien。在此基础上,将合成的二种氨基化-β-环糊精用于手性拆分中性邻二醇对映体,取得了良好的手性分离效果,其手性识别能力明显优于天然中性β-环糊精,特别是对端基手性邻二醇表现出更加独特的手性识别作用。以β-CDen为优选的手性选择剂,建立了毛细管电泳分离分析8种手性邻二醇的新方法,在优化条件下8种手性邻二醇均得到基线分离。用建立的方法测定了8种不对称二羟化反应产物的百分对映体过量值（%ee）,并与HPLC方法的测定结果比较,二者完全一致。3.以高磺化-β-CD作为毛细管电泳手性选择剂,建立了有效分离7种手性芳香仲醇对映体的新方法,在优化的电泳条件下,所有芳香仲醇对映体均实现了基线分离。用所建立的方法测定了芳香酮不对称氢转移反应产物芳香仲醇的百分对映体过量值（%ee）,其测定结果与GC和HPLC测定结果完全一致。4.以天然L-（+）-酒石酸为原料,经酯化、羟基保护、还原、与MsCl酯化、叠氮化、催化还原等六步反应合成了（2S,3S）-（+）-2,3-O-异丙叉-1,4-丁二胺。再以（1R,2R）-（-）-1,2-二氨基环己烷和（2S,3S）-（+）-2,3-O-异丙叉-1,4-丁二胺为原料,在天然β-环糊精结构中引入二类不同的手性基因中心,分别合成了二种新的阳离子型手性胺-β-环糊精手性选择剂,将二者用于毛细管电泳手性拆分阴离子化合物。在100mmol·L^-1 NaH₂PO₄-Na₂HPO₄缓冲体系中,20℃柱温,15kV操作电压下,对10种丹酰化氨基酸和N-乙酰苯丙氨酸对映体进行了手性拆分,取得了良好的分离效果,同时,对4种氨基酸混合样品进行分析,亦取得了满意的对映体分离。更多还原

【Abstract】 In recent years, capillary electrophoresis （CE） has become one of the rapid development branch of chiral analysis due to its unique features, such as simplification, high efficiency, sensitiveness, trace consumption of sample, solvent and chiral selectors. CE has been widely used in life science, chemistry and pharmaceutical sciences, especially in chiral resolution and analysis of pharmaceutical and biomacromolecule. In CE chiral separation, the chiral selector is usually dissolved in the running buffer to provide chiral environment. The difference of the interactions between chiral selectors and enantiomers leads to the difference of electrophoretic mobilities, so the two enantiomers can be separated.In this thesis, four chiral selectors have been synthesized and used in CE for the resolution of chiral vicinal diols, chiral secondary aromatic alcohols and amino acids derivatives, respectively. Moreover, the mechanism of chiral resolution has been discussed.This thesis mainly focuses on four aspects as follows:1. A new access was improved to synthesis of 1,4-bis （9-O-quininyl） phthalazine [（QN）2PHAL], a very useful chiral ligand for catalytic asymmetric dihydroxyl- ation （AD）, by using CaH₂ as acid-binding reagent in a high yield under mild conditions. The application of （QN）2PHAL to the AD reactions of eight olefins exhibited excellent enantioselectivity and activity with corresponding chiral vicinal diols. Furthermore, a capillary zone electrophoresis （CZE） method was developed to separate the synthetic chiral vicinal diols by using neutralβ-cyclodextrin（β-CD）as chiral selector and borate as running buffer. High resolution was achieved under the optimal conditions (β-CD 2.2% （w/v）, pH 10, 200 mmol·L^-1 borate buffer at 15 kV and 20℃within 15 min). The relative standard deviations of the corrected peak areas and migration time were less than 3.9% and 1.3%, respectively. In addition, the developed method was successfully applied to the determination of the purity and the enantiomeric excesses value （%ee） of the AD reaction products.2. Two new kinds of cationic amino-β-cyclodextrin chiral selector （β-CDen andβ-CDdien） have been prepared via modification of C6-β-cyclodextrin. The synthetic method to prepare intermediate （6-OTs-β-CD） was also improved in higher yield and simpler procedure.β-CDen andβ-CDdien were obtained by the reaction of 6-OTs-β-CD and ethylenediamine or diethylenetriamine in the presence of trace ammount of DMAP, and the chemical yields was 50.3% and 51.6%, respectively. Furthermore, these new chiral selectors were successfully applied for capillary electrophoresis separations of enantiomers of eight vicinal diols. To our delight, these new chiral selectors have achieved good results in chiral separation. The chiral recognition ability is superior to the naturalβ-neutral cyclodextrin, especially for the terminal chiral vicinal diols, showing greater chiral recognition. High resolution was achieved under the optimized conditions ofβ-CDen 10mmol·L^-1, pH 10, 200 mmol·L^-1 borate buffer at 15 kV and 20℃. In addition, the established method was successfully applied to the determination of the enantiomeric excesses value （%ee） of the AD reaction products. It was also found that the results of the CE analysis was in accordance with that of HPLC, which indicates that the proposed method is suitable for the determination of %ee value of the vicinal diols samples.3. A new CE method was developed to efficiently separate seven chiral secondary aromatic alcohols by using S-β-CD as chiral selector. Under the optimal conditions, the baseline separation of the enantiomers of secondary aromatic alcohols was obtained. In addition, the established method was successfully applied to the determination of the enantiomeric excesses value （%ee） of products of asymmetric transfer hydrogenation reactions. It was found that the results of determination by the CE analysis were in accordance with that by GC and HPLC.4. （2S,3S）-（+）-2,3-O-isopropylidene-l,4-tetramethylenediamine was synthesized in six steps from （2R,3R）-tartaric acid via esterification, hydroxyl protection, reduction, and MsCl esterification, azide reaction, and catalytic hydrogenation. Furthermore, two kinds of new cationic chiral amine-β-cyclodextrin chiral selector, which was used to enantioseparate the anionic chiral compounds, were synthesized from （1R,2R）-（-）-1,2-diaminocyclohexane and （2S,3S）-（+）-2,3-O- isopropylidene-l,4-tetramethylenediamine by introducing two chiral centre in C6 ofβ-CD, respectively. Furthermore, a CE method was established to separate 10 Dns-amino acids and N-acetylphenylalanine by using above-mentioned synthetic aminoderivatives ofβ-CD as chiral selector and phosphate as running buffer. Excellent enantiomeric separation was obtained under the optimal conditions (chiral selector 10mmol·L^-1, pH 6.5, 20 mmol·L^-1 Na2HPO4-NaH₂PO4 buffer at 15 kV and 20℃). Moreover, satisfactory enantioseparation was also achieved for the mixed samples of four amino acids under the optimal conditions.更多还原