【摘要】：目的：本文探討了β-環(huán)糊精和手性離子液體聯(lián)用，以及手性配體交換法拆分苯丙氨酸（Phe）、色氨酸（Trp）和酪氨酸（Tyr）對(duì)映體的毛細(xì)管電泳的分離機(jī)理；建立簡(jiǎn)單、快速、綠色的高效毛細(xì)管電泳法，并用于測(cè)定氨基酸注射液，口服液以及人體尿液中氨基酸的含量。 方法：(1)以β-環(huán)糊精和手性離子液體聯(lián)用為分離介質(zhì)體系，建立拆分苯丙氨酸和色氨酸對(duì)映體的毛細(xì)管電泳新方法。本文分別考察了緩沖液濃度、pH、β-CD濃度、手性離子液體（CIL）濃度和分離電壓，得到測(cè)定注射液中氨基酸含量的最佳毛細(xì)管電泳條件為：未涂層毛細(xì)管柱（75μm×57cm，有效長(zhǎng)度50cm）、緩沖液15mM硼砂、pH9.5、5mM β-CD、4mM CIL和分離電壓10kV，檢測(cè)波長(zhǎng)214nm，壓力進(jìn)樣:0.5psi，3s。(2)建立以Cu2+和L-脯氨酸（L-Pro）為手性添加劑的配體交換法分離氨基酸對(duì)映體的毛細(xì)管電泳方法，并應(yīng)用于實(shí)際樣品中氨基酸的含量測(cè)定。文中考察了緩沖液濃度、SDS的濃度、pH、Cu2+和L-Pro的比例以及二者的濃度和分離電壓，得到最佳的分離和測(cè)定條件為：未涂層毛細(xì)管柱（75μm×57cm，有效長(zhǎng)度50cm）、緩沖液10mM醋酸銨、30mM SDS、pH5.0、10mM Cu(Au)2、20mM L-Pro和分離電壓15kV，檢測(cè)波長(zhǎng)214nm，壓力進(jìn)樣:0.5psi，3s。 結(jié)果：(1)在最佳毛細(xì)管電泳條件下，測(cè)得分析物線性范圍為0.08~10μg/mL；相關(guān)系數(shù)R2為0.9956~0.9998；在6min內(nèi)完成出峰；分離度在1.57~3.22之間；日內(nèi)RSD＜4.3%，日間RSD＜7.2%。樣品中分析物的回收率在88.1%~115.8%之間。(2)在最佳毛細(xì)管電泳條件下，測(cè)得D，L-Tyr、D，L-Trp和L-Phe、D-Phe線性范圍分別為2~150μg/mL，0.5~100μg/mL，0.1~100μg/mL，0.1~80μg/mL；相關(guān)系數(shù)R2為0.9917~0.9984；分離度在1.82~2.48之間；日內(nèi)RSD＜5.1%,日間RSD＜5.7%。樣品中分析物的回收率在85.6%~115.2%之間。 結(jié)論：手性離子液體作為手性選擇性試劑可用于氨基酸的手性分離；手性配體交換毛細(xì)管電泳法分離氨基酸時(shí)簡(jiǎn)單，，快速。本文所建立的的毛細(xì)管電泳新方法可應(yīng)用于實(shí)際樣品分析。
[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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R917

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