【作者】 孙苏亚；

【导师】 李发美；

【作者基本信息】 沈阳药科大学 ， 药物分析， 2006， 博士

【摘要】 手性配体交换色谱法是分离手性化合物的基本方法之一，其中手性配体和金属离子是决定手性化合物分离选择性的根本因素。本文采用流动相添加法和固定相涂渍法对手性化合物对映体进行了分离，并研究了对映体分离中的多种影响因素，探讨了其手性分离机制。研究了手性配体与二价金属离子Fe²⁺、Co²⁺、Ni²⁺、Zn²⁺、Cu²⁺、Pb²⁺、Hg²⁺所形成配合物的稳定性及它们在手性分离中的重要作用。 1．首次采用流动相添加法定量分析了盐酸贝那普利和八氢吲哚-2-羧酸及它们的异构体，对金属离子、配体、离子对试剂、pH、有机溶剂、柱温等多种因素对保留行为和分离度的影响进行了考察，并对其色谱行为及机制进行了探讨，优化了色谱条件。分离八氢吲哚-2-羧酸异构体的色谱条件为：以Zorbax bonus-C₈为固定相，以含L-PheA 2.0mmol／L、醋酸铜1.0mmol／L、辛烷磺酸钠5.0mmol／L的水溶液（调pH为6.0）-乙腈（80:20，v／v）为流动相。分离贝那普利异构体的色谱条件为：以Diamonsil C₁₈为固定相，流动相为含L-PheA 2.0mmol／L、醋酸铜1.0mmol／L、辛烷磺酸钠2.0mmol／L的水溶液-甲醇（40:60，v／v）。八氢吲哚-2-羧酸和盐酸贝那普利的异构体均得到完全分离，分别在10～300μg·mL^-1和5～300μg·mL^-1浓度范围内峰面积与浓度呈良好的线性关系。该方法简便经济、定量准确、重现性好。 在手性配体交换色谱法中添加不同浓度的辛烷磺酸钠，观察其对氨基酸对映体分离的影响，研究了辛烷磺酸钠在手性配体交换色谱法中的作用。结果表明，辛烷磺酸钠可以增强氨基酸的保留时间，尤其是D型氨基酸在色谱柱上的保留时间。 2．采用简单的合成工艺合成了五种手性配体：N-十二烷基-L-羟基脯氨酸（C12-L-Hyp）、N-辛烷基-L-羟基脯氨酸（C8-L-Hyp）、N-辛烷基-L-脯氨酸（C8-L-Pro）、N-对甲苯磺酸-L-羟基脯氨酸和L-苯丙酰胺。N-对甲苯磺酸-L-羟基脯氨酸和L-苯丙酰胺的合成产品纯度＞98％，收率均达到80％以上。并在随后的研究中将这些配体用于手性配体交换色谱法。 3．采用固定相涂渍法，研究了C12-L-Hyp、C8-L-Hyp、C8-L-Pro、N-对甲苯磺酸-L-羟基脯氨酸和（S）-3-氨基-2，3，4，5-四氢-1H-[1]-苯并氮杂卓-2-酮-1-乙酸叔丁酯（TBAB）更多还原

【Abstract】 Chiral ligand-exchange chromatography （CLEC） is one of the basic chromatography method for enantiomer separation, in which chiral ligands and metal ions are two essential elements to determine the chiral selectivity. In this study enantiomers of some chiral drugs and amino acids were separated, by chiral coated phases （CCP） and chiral mobile phases （CMP）. Various factors affecting the chromatographic behaviour and chiral selectivity and mechanism were studied. The stability of complexes between chiral ligands and metal ions (Fe²⁺、Co²⁺、Ni²⁺、Zn²⁺、Cu²⁺、Pb²⁺、Hg²⁺) and the critical role in chiral separation were studied by using CCP and CMP methods.1. The isomers of Octahydroindole-2-carboxylic acid （Oic） and Benazepril Hydrochloride（BNP） were separated and assayed by using CMP. The factors such as metal ions, ligands, ion-pair reagents, pH, column temperature and organic modifiers affecting the retention and resolution were studied. The isomers of Oic were separated on Zorbax bonus-C₈ column with the mobile phase of 20% acetonitrile and 80% aqueous solution （pH 6.0） containing 1.0 mmol/L Cu （II） acetate, 2.0 mmol/L L-phenylalaninamide （L-PheA） and 5.0 mmol/L PIC-B. The isomers of BNP were separated on Diamonsil C₁₈ column with the mobile phase of 60% methanol and 40% aqueous solution （pH 6.0） containing 1.0 mmol/L Cu （II） acetate, 2.0 mmol/L Vitasweet （APM） and 2.0 mmol/L PIC-B. Baseline separation of the isomers of Oic and BNP were obtained under these chromatographic conditions, and a linear relationship between peak area and concentration was achieved in the range of 10～ 300μg·mL^-1 and 5～300 μg·mL^-1, respectively.The effect concentration of ion-pair reagent on retention factors and resolution of amino acids in CMP was studied. The results indicated that the retention time of amino acids especially D-amino acids were increased when PIC-B8 were added in mobile phase.2. Some chiral ligands deriving from amino acids, N-（n-dodecyl）-L-hydroxyproline更多还原