毛细管电泳技术在喜树生物碱分离分析中的应用研究

发布时间：2018-03-08 23:12

本文选题：喜树生物碱　切入点：毛细管电泳法　出处：《山西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:本文建立了以磺丁基-β-环糊精(SBE-β-CD)和离子液体(IL)联合作为添加剂分离分析喜树生物碱的毛细管电泳法,并且研究了样品在线堆积的富集方法及其在毛细管电泳分析测定喜树生物碱中的应用;所建立的方法简单、方便、高效且节省有机溶剂,可用于分离测定实际样品喜树果和喜树皮中的喜树生物碱。方法:(1)以SBE-β-CD和IL联合作为毛细管电泳背景电解质添加剂,建立了分离喜树生物碱的新方法,并用于实际样品中喜树碱的分离测定。通过考察不同浓度的缓冲溶液、p H、SBE-β-CD和IL浓度、分离电压等对喜树生物碱分离的影响,得到分离分析喜树生物碱的最佳毛细管电泳条件为:未涂层熔融毛细管(50 cm×75μm,有效长度40 cm);缓冲溶液为:20 m M的硼砂溶液,20 m M SBE-β-CD,100 m M IL(p H9.0);分离电压为15 k V,进样条件为:0.5 psi压力进样4.0 s,检测波长为254 nm。(2)建立了毛细管电泳大体积样品堆积法(LVSS)分析测定微量喜树生物碱的新方法,并以SBE-β-CD和IL联合作为背景电解质添加剂来分离喜树生物碱,并将其用于实际样品喜树果和喜树皮中喜树生物碱的分离和含量测定。本文考察了缓冲溶液的浓度和p H、SBE-β-CD和离子液体(IL)浓度、分离电压、堆积条件等的影响,优化得到了大体积样品堆积毛细管电泳法的最佳条件为:未涂层熔融毛细管(50cm×75μm,有效长度40 cm);最佳分离条件为25 m M的硼砂溶液(p H 9.0),20 m M SBE-β-CD,20 m M IL,分离电压20 k V,检测波长254 nm;最佳堆积条件为:进样压力0.5 psi,进样时间4.0 s,反向电压-25 k V,电极反转时间0.17 min。结果:(1)在最佳的电泳条件下,喜树生物碱可以在10 min内完成分离;日内精密度RSD4.6%,日间精密度RSD7.6%;各分析物的线性相关系数R为0.9946~0.9985;实际样品中各分析物的平均回收率为98.8%~103.6%。(2)在最佳的毛细管电泳条件下,五种喜树生物碱可以在20 min内完成分离,日内精密度RSD1.23%,日间精密度RSD3.70%;各分析物的线性相关系数R在0.9991~0.9997之间;在最佳的大体积样品堆积条件下,分析物的富集倍数可达到5~13倍不等;实际样品中各分析物的平均回收率为95.0%~103.2%。结论:离子液体可与环糊精联合作为毛细管电泳背景缓冲液的添加剂,用于喜树生物碱的分离测定;大体积样品堆积法可用于实际样品中含量较低的喜树生物碱的分析测定。毛细管电泳技术为喜树生物碱的分离与测定提供了一种更经济、快速、环保的新方法,离子液体与环糊精可作为分离选择性试剂应用于毛细管电泳分离喜树生物碱的背景电解质体系中。
[Abstract]:Objective: to establish a capillary electrophoresis method for the separation and analysis of camptothecin with sulfon#china_person0#-尾 -cyclodextrin (SBE- 尾 -CD) and ionic liquid (ILL) as additives. The method of sample accumulation on line and its application in the determination of camptothecin by capillary electrophoresis were studied. The method is simple, convenient, efficient and organic solvent saving. It can be used for the separation and determination of camptothecin in actual samples and bark of Camptotheca acuminata. Methods A new method for separation of camptothecin from Camptotheca acuminata was established by using SBE- 尾 -CD and IL as background electrolyte additives in capillary electrophoresis. It was applied to the separation and determination of camptothecin in practical samples. The effects of different concentrations of buffer solution, such as the concentration of SBE- 尾 -CD and IL, the separation voltage, on the separation of camptothecin were investigated. The optimum conditions for separation and analysis of camptothecin were obtained as follows: uncoated fused capillary tube 50 cm 脳 75 渭 m, effective length 40 cm ~ (-1); borax solution: 20 m M SBE- 尾 -CD1 100mm IL(p H 9.0m buffer solution; separation voltage 15 kV, sample injection. A new method for the determination of trace camptothecin was established by mass sample stacking method in capillary electrophoresis (Capillary electrophoresis), under the condition that the sample was injected under the pressure of 1: 0.5 psi for 4.0 s and the detection wavelength was 254nm.m-2). SBE- 尾 -CD and IL were used as background electrolyte additives to isolate camptotheca alkaloids. It has been applied to the separation and determination of camptothecin in actual samples and bark of Camptotheca acuminata. The effects of the concentration of buffer solution, the concentration of pHHSBE- 尾 -CD and ionic liquid (ILL), the separation voltage, the stacking conditions, etc. The optimum conditions for bulk sample stacking capillary electrophoresis were obtained as follows: uncoated fused capillary tube 50 cm 脳 75 渭 m, effective length 40 cm ~ (-1), borax solution p H 9.0 ~ 20 m ~ (-1), SBE- 尾 -CD ~ (2 +) 20 mm IL, separation voltage 20 k. V, the detection wavelength is 254 nm, the optimum packing conditions are as follows: injection pressure 0.5 psil, injection time 4.0 s, reverse voltage -25 kV, electrode reversal time 0.17 min. Camptothecin can be separated within 10 min; intra day precision RSD 4.6, day precision RSD 7.6; linear correlation coefficient R = 0.9946 ~ 0.9985; average recovery rate of analysis is 98.8 / 103.6. under the optimum conditions of capillary electrophoresis, The five camptothecins can be separated in 20 min, the intra day precision is 1.23 and the daytime precision is 3.70. The linear correlation coefficient of each analyte is between 0.9991 and 0.9997. The average recovery of each analyte was 95.0 and 103.2.Conclusion: ionic liquid and cyclodextrin can be used as additives in background buffer of capillary electrophoresis for the separation and determination of camptothecin. The bulk sample stacking method can be used for the analysis and determination of camptothecin in practical samples. Capillary electrophoresis provides a more economical, rapid and environmentally friendly method for the separation and determination of camptothecin. Ionic liquids and cyclodextrin can be used as separation selective reagents in the background electrolyte system for separation of camptotheca alkaloids by capillary electrophoresis.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O658.9;R914

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