基于手性离子液体的毛细管电泳技术在手性氨基酸分离中的应用研究

发布时间：2019-04-22 13:57

【摘要】：目的：本文探讨了β-环糊精和手性离子液体联用，以及手性配体交换法拆分苯丙氨酸（Phe）、色氨酸（Trp）和酪氨酸（Tyr）对映体的毛细管电泳的分离机理；建立简单、快速、绿色的高效毛细管电泳法，并用于测定氨基酸注射液，口服液以及人体尿液中氨基酸的含量。方法：(1)以β-环糊精和手性离子液体联用为分离介质体系，建立拆分苯丙氨酸和色氨酸对映体的毛细管电泳新方法。本文分别考察了缓冲液浓度、pH、β-CD浓度、手性离子液体（CIL）浓度和分离电压，得到测定注射液中氨基酸含量的最佳毛细管电泳条件为：未涂层毛细管柱（75μm×57cm，有效长度50cm）、缓冲液15mM硼砂、pH9.5、5mM β-CD、4mM CIL和分离电压10kV，检测波长214nm，压力进样:0.5psi，3s。(2)建立以Cu2+和L-脯氨酸（L-Pro）为手性添加剂的配体交换法分离氨基酸对映体的毛细管电泳方法，并应用于实际样品中氨基酸的含量测定。文中考察了缓冲液浓度、SDS的浓度、pH、Cu2+和L-Pro的比例以及二者的浓度和分离电压，得到最佳的分离和测定条件为：未涂层毛细管柱（75μm×57cm，有效长度50cm）、缓冲液10mM醋酸铵、30mM SDS、pH5.0、10mM Cu(Au)2、20mM L-Pro和分离电压15kV，检测波长214nm，压力进样:0.5psi，3s。结果：(1)在最佳毛细管电泳条件下，测得分析物线性范围为0.08~10μg/mL；相关系数R2为0.9956~0.9998；在6min内完成出峰；分离度在1.57~3.22之间；日内RSD＜4.3%，日间RSD＜7.2%。样品中分析物的回收率在88.1%~115.8%之间。(2)在最佳毛细管电泳条件下，测得D，L-Tyr、D，L-Trp和L-Phe、D-Phe线性范围分别为2~150μg/mL，0.5~100μg/mL，0.1~100μg/mL，0.1~80μg/mL；相关系数R2为0.9917~0.9984；分离度在1.82~2.48之间；日内RSD＜5.1%,日间RSD＜5.7%。样品中分析物的回收率在85.6%~115.2%之间。结论：手性离子液体作为手性选择性试剂可用于氨基酸的手性分离；手性配体交换毛细管电泳法分离氨基酸时简单，，快速。本文所建立的的毛细管电泳新方法可应用于实际样品分析。
[Abstract]:Objective: to investigate the mechanism of separation of enantiomers of phenylalanine (Phe), tryptophan (Trp) and tyrosine (Tyr) by chiral ligand exchange coupled with 尾 cyclodextrin (尾 cyclodextrin) and chiral ionic liquids (ionic liquids). A simple, rapid and green capillary electrophoresis method was developed for the determination of amino acid in amino acid injection, oral liquid and human urine. Methods: (1) A new capillary electrophoresis method for separation of phenylalanine and tryptophan enantiomers was established by using 尾-cyclodextrin and chiral ionic liquids as separation medium. In this paper, the buffer concentration, pH, 尾-CD concentration, (CIL) concentration and separation voltage of chiral ionic liquids were investigated respectively. The optimum capillary electrophoresis conditions for the determination of amino acids in injection were obtained as follows: uncoated capillary column (75 渭 m 脳 57 cm, P < 0.05). Effective length 50cm), buffer 15mM borax, pH9.5,5mM 尾-CD,4mM CIL and separation voltage 10 kV, detection wavelength 214 nm, pressure injection: 0.5 psi, 3. (2) A capillary electrophoresis method for separation of enantiomers of amino acids using Cu2 and L-proline (L-Pro) as chiral additives was established and applied to the determination of amino acids in practical samples. The concentration of buffer, the concentration of SDS, the ratio of pH,Cu2 and L-Pro, the concentration of both and the separation voltage were investigated. The optimum separation and determination conditions were obtained as follows: uncoated capillary column (75 渭 m 脳 57 cm, effective length 50cm). Buffer 10mM ammonium acetate, 30mM SDS,pH5.0,10mM Cu (Au) 2, 20mm L-Pro and separation voltage 15kV, detection wavelength 214 nm, pressure injection: 0.5psi, 3s. Results: (1) under the optimum conditions of capillary electrophoresis, the linear range of the analyte was 0.08 ~ 10 渭 g / mL;, the correlation coefficient R2 was 0.9956 渭 g / 6min, the peak was achieved in 6min, the resolution was between 1.57 and 3.22, and the correlation coefficient was 0.999 6 脳 10 渭 g / mol. Intraday RSD < 4.3%, daytime RSD < 7.2%. The recoveries of the analytes were between 88.1% and 115.8%. (2) under the optimum conditions of capillary electrophoresis, the linear ranges of D, L, L and H were 2 ~ 150 渭 g / mL,0.5~ 100 渭 g / mL, respectively, and the linear range was 2 ~ 150 渭 g / mol ~ (- 1) 100 渭 g / mL,. 0.1 ~ 100 渭 g / mL,0.1~ 80 渭 g / mL; The correlation coefficient (R2) was 0.9917 and 0.9984, the resolution was between 1.82 and 2.48, the intra-day RSD was < 5.1%, the inter-day RSD was less than 5.7%. The recoveries of the analytes in the samples were between 85.6% and 115.2%. Conclusion: chiral ionic liquids can be used as chiral selective reagents for chiral separation of amino acids, and chiral ligand exchange capillary electrophoresis is a simple and rapid method for the separation of amino acids. The new capillary electrophoresis method developed in this paper can be applied to the analysis of practical samples.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R917

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