毛细管电泳—电致化学发光法在环境及药物分析中的应用研究
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摘要
毛细管电泳( Capillary electrophoresis, CE )电致化学发光(Electrochemiluminescence, ECL)联用技术兼备分离效率高和灵敏度高等特点,在药物、生物、临床、食品分析等领域得到广泛的应用。但将CE-ECL技术用于环境分析领域的研究报道很少。此外,生命体液中的干扰杂质对裸铂电极的吸附毒化会严重降低检测的灵敏度和稳定性,因此本工作的主要内容是利用实验小组制备的稀土铕离子(III)掺杂的类普鲁士蓝膜(Eu-PB)修饰的铂电极作为工作电极,建立了一系列基于Ru(bpy)32+电致化学发光反应的CE-ECL分析方法,实现了土壤中吸附的多抗霉素B、药物制剂及尿样中的富马酸酮替芬以及兔血浆中的氢溴酸高乌甲素和富马酸异丙吡仑的分离检测,有助于进一步拓展环境物质的分离分析方法以及扩大CE-ECL分析方法的应用范围。
论文包括以下四章:
第一章文献综述
介绍了CE技术和ECL分析的基本原理和基础理论,对其联用技术的特点、常用体系及在环境和药物中的应用进行了归纳总结,展望了CE-ECL分析的发展趋势及应用前景,共引用文献116篇。
第二章毛细管电泳-电致化学发光法测定土壤中的多抗霉素B
以稀土铕离子(III)掺杂的类普鲁士蓝膜(Eu-PB)修饰铂电极为工作电极,采用毛细管电泳-电致化学发光法(CE-ECL)对土壤中的多抗霉素B进行检测。分别对毛细管电泳分离条件和电致化学发光检测条件进行了优化,并探讨了体系产生电致化学发光的机理。在优化实验条件下,多抗霉素B可在4 min内得到分离,其ECL强度值与多抗霉素B浓度的对数值在1.0×10~(-7)~5.0×10~(-7) g/mL和5.0×10~(-7)~5.0×10~(-5) g/mL范围内呈良好的线性关系,检出限(3σ)为9.2×10~(-8) g/mL。对5.0×10-6 g/mL多抗霉素B溶液进行平行测定,ECL强度和迁移时间的日内及日间RSD值均小于5 %。将该法用于膨润土、高岭土、黄土3种土壤中多抗霉素B含量的测定,加标回收率在92.2 %~104.3 %之间;多抗霉素B在3种土壤中吸附2 h后的吸附率分别为89.0 %、45.5 %和40.4 %,表明3种土壤对多抗霉素B的吸附能力依次为:膨润土>高岭土>黄土,其原因可能与土壤自身的特性、比表面积及阳离子交换容量(CEC)有关。
第三章毛细管电泳-电致化学发光法测定药物制剂及尿样中的富马酸酮替芬
基于稀土铕离子(III)掺杂的类普鲁士蓝膜修饰的铂电极为工作电极,建立了测定富马酸酮替芬的毛细管电泳-电致化学发光分析的新方法。考察了检测电位、运行缓冲溶液的酸度及浓度、分离电压、进样条件等实验参数对富马酸酮替芬测定的影响。在最佳条件下,ECL强度与富马酸酮替芬的浓度在3.0×10~(-8)~2.0×10~(-6) g/mL和2.0×10~(-6)~5.0×10~(-6) g/mL范围内呈良好的线性关系,检出限(3σ)为2.1×10~(-8) g/mL。对4.0×10~(-7) g/mL的富马酸酮替芬平行测定5次,峰高和迁移时间的相对标准偏差分别为0.95 %和0.26 %。该法简便、快速、灵敏,用于药物制剂及尿样中富马酸酮替芬的测定,加标回收率在99.5 %~107.0 %之间。
第四章毛细管电泳-电致化学发光法同时测定兔血浆中的氢溴酸高乌甲素和富马酸异丙吡仑
采用毛细管电泳-电致化学发光检测法对镇痛药氢溴酸高乌甲素和富马酸异丙吡仑进行快速、灵敏的分离检测。考察了检测电位、运行缓冲液浓度与pH值、分离电压、进样电压、进样时间等实验条件对分离检测的影响。在最佳条件下,氢溴酸高乌甲素和富马酸异丙吡仑在6 min内可实现分离检测,ECL强度与氢溴酸高乌甲素和富马酸异丙吡仑的浓度分别在1.0×10~(-7)~5.0×10~(-6) g/mL和5.0×10~(-6)~5.0×10~(-5) g/mL、4.0×10~(-8)~5.0×10~(-7) g/mL和5.0×10~(-7)~1.0×10~(-5) g/mL范围内呈良好的线性关系,检出限(3σ)分别为6.6×10~(-8) g/mL和3.7×10~(-8) g/mL。该方法已成功用于兔血浆中氢溴酸高乌甲素和富马酸异丙吡仑的同时检测,加标回收率在95.6 %~103.0 %之间。
Capillary electrophoresis(CE) with electrochemiluminescence (ECL) detection, as an high-efficient and high-sensitive analytical technique, has been extensively applied in the fields of pharmaceutical, biochemical, clinical and food analysis. But it is a pity that only a few studies on assay of environmental analytes have been reported by CE-ECL detection. On the other hand, the sensitivity and stability of a CE-ECL method will be greatly decreased since the surface of bare platinum working electrode is often contaminated by adsorbing sample matrices in biological fluids. Therefore, in this paper, a microdisk platinum electrode modified with europium (III)-doped prussian blue analogue (Eu-PB) film was used as a working electrode. Given this, a series of Ru(bpy)32+-based ECL detection coupled with CE have been established for the determination of polyoxin B absorbed in soils, ketotifen fumarate in pharmaceuticals and human urine, and lappaconitine hydrobromide, isopropiram fumarate in rabbit plasma. The present work would be helpful to the further development of detection methods for some environmental analytes and extend applications of CE-ECL technique.
This paper consists of four chapters:
Chapter One Review
The basic principles of CE and ECL are introduced firstly, and then the characteristics, the common systems and the application of CE-ECL technique in the fields of environmental and pharmaceutical analysis are summarized. Finally, the prospect of the newly developing techniques and further trends of CE-ECL methods are reviewed, and 116 references are cited.
Chapter Two Determination of polyoxin B in soils by capillary electrophoresis - electrochemiluminescence detection
On the basis of a chemically modified platinum electrode by europium (III)-doped prussian blue analogue film (Eu-PB) as a working electrode, a rapid and sensitive method for determination of polyoxin B was established using CE-ECL detection. Parameters related to the separation and detection were discussed and optimized. Under the optimized conditions, polyoxin B was separated within 4 min, and the ECL intensity was in proportion to polyoxin B concentration over the range of 1.0×10~(-7)~5.0×10~(-7) g/mL and 5.0×10~(-7)~5.0×10~(-5) g/mL with a detection limit (3σ) of 9.2×10-8 g/mL. The relative standard deviations (RSD) of intra-day and inter-day ECL intensity and migration time were less than 5.0 %. The proposed method was successfully applied in the determination of polyoxin B absorbed in three kinds of soils (e.g. bentonite, kaolinite and loess) with the recoveries of 92.2 % ~104.3 %. The results also showed that the adsorption rate of polyoxin B in soils for 2 h was 89.0 % for bentonite, 45.5 % for kaolinite and 40.4 % for loess, respectively, indicating that the adsorption capacity of polyoxin B in different soils was bentonite> kaolinite>loess. The possible reason was related to the characteristics, specific surface area and cation exchange capacity (CEC) of soils.
Chapter Three Determination of ketotifen fumarate in pharmaceuticals and human urine by capillary electrophoresis - electrochemiluminescence detection
On the basis of a chemically modified platinum electrode by europium (III)-doped prussian blue analogue film (Eu-PB) as a working electrode, a simple, rapid and sensitive CE-ECL method was developed for the determination of ketotifen fumarate (KTF). The effects of several factors such as the applied potential, the acidity and the concentration of running buffer, separation voltage, and injection conditions on the detection were investigated. Under the optimized conditions, the ECL intensity was in proportion to KTF concentration over the range of 3.0×10-8~2.0×10~(-6) g/mL and 2.0×10~(-6)~5.0×10~(-6) g/mL with a detection limit (3σ) of 2.1×10-8 g/mL. The relative standard deviations of the ECL intensity and the migration time were 0.95 and 0.26 %, respectively. The developed method was successfully applied to the determination of KTF contents in pharmaceuticals and human urine with the recoveries between 99.5 and 107.0 %.
Chapter Four Simultaneous determination of lappaconitine hydrobromide and isopropiram fumarate in rabbit plasma by capillary electrophoresis - electrochemiluminescence detection
A rapid and sensitive method for simultaneous determination of lappaconitine hydrobromide (LH) and isopropiram fumarate (IF) has been firstly established by CE - ECL detection. The effect of analytical conditions, such as applied potential, the pH and concentration of running buffer,separation voltage, injection voltage and injection time were investigated in detail. Under the optimized conditions, a baseline separation for the two analytes was achieved within 6 min, and the ECL intensity was in proportion to LH and IF concentration over the range of 1.0×10~(-7)~5.0×10~(-6) g/mL and 5.0×10~(-6)~5.0×10~(-5) g/mL for LH and 4.0×10~(-8)~5.0×10~(-7) g/mL and 5.0×10~(-7)~1.0×10~(-5) g/mL for IF with a detection limit (3σ) of 6.6×10~(-8) g/mL for LH and 3.7×10~(-8) g/mL for IF, respectively. The applicability of the proposed method was illustrated for the determination of LH and IF in rabbit plasma with recoveries between 95.6 and 103.0 %.
论文包括以下四章:
第一章文献综述
介绍了CE技术和ECL分析的基本原理和基础理论,对其联用技术的特点、常用体系及在环境和药物中的应用进行了归纳总结,展望了CE-ECL分析的发展趋势及应用前景,共引用文献116篇。
第二章毛细管电泳-电致化学发光法测定土壤中的多抗霉素B
以稀土铕离子(III)掺杂的类普鲁士蓝膜(Eu-PB)修饰铂电极为工作电极,采用毛细管电泳-电致化学发光法(CE-ECL)对土壤中的多抗霉素B进行检测。分别对毛细管电泳分离条件和电致化学发光检测条件进行了优化,并探讨了体系产生电致化学发光的机理。在优化实验条件下,多抗霉素B可在4 min内得到分离,其ECL强度值与多抗霉素B浓度的对数值在1.0×10~(-7)~5.0×10~(-7) g/mL和5.0×10~(-7)~5.0×10~(-5) g/mL范围内呈良好的线性关系,检出限(3σ)为9.2×10~(-8) g/mL。对5.0×10-6 g/mL多抗霉素B溶液进行平行测定,ECL强度和迁移时间的日内及日间RSD值均小于5 %。将该法用于膨润土、高岭土、黄土3种土壤中多抗霉素B含量的测定,加标回收率在92.2 %~104.3 %之间;多抗霉素B在3种土壤中吸附2 h后的吸附率分别为89.0 %、45.5 %和40.4 %,表明3种土壤对多抗霉素B的吸附能力依次为:膨润土>高岭土>黄土,其原因可能与土壤自身的特性、比表面积及阳离子交换容量(CEC)有关。
第三章毛细管电泳-电致化学发光法测定药物制剂及尿样中的富马酸酮替芬
基于稀土铕离子(III)掺杂的类普鲁士蓝膜修饰的铂电极为工作电极,建立了测定富马酸酮替芬的毛细管电泳-电致化学发光分析的新方法。考察了检测电位、运行缓冲溶液的酸度及浓度、分离电压、进样条件等实验参数对富马酸酮替芬测定的影响。在最佳条件下,ECL强度与富马酸酮替芬的浓度在3.0×10~(-8)~2.0×10~(-6) g/mL和2.0×10~(-6)~5.0×10~(-6) g/mL范围内呈良好的线性关系,检出限(3σ)为2.1×10~(-8) g/mL。对4.0×10~(-7) g/mL的富马酸酮替芬平行测定5次,峰高和迁移时间的相对标准偏差分别为0.95 %和0.26 %。该法简便、快速、灵敏,用于药物制剂及尿样中富马酸酮替芬的测定,加标回收率在99.5 %~107.0 %之间。
第四章毛细管电泳-电致化学发光法同时测定兔血浆中的氢溴酸高乌甲素和富马酸异丙吡仑
采用毛细管电泳-电致化学发光检测法对镇痛药氢溴酸高乌甲素和富马酸异丙吡仑进行快速、灵敏的分离检测。考察了检测电位、运行缓冲液浓度与pH值、分离电压、进样电压、进样时间等实验条件对分离检测的影响。在最佳条件下,氢溴酸高乌甲素和富马酸异丙吡仑在6 min内可实现分离检测,ECL强度与氢溴酸高乌甲素和富马酸异丙吡仑的浓度分别在1.0×10~(-7)~5.0×10~(-6) g/mL和5.0×10~(-6)~5.0×10~(-5) g/mL、4.0×10~(-8)~5.0×10~(-7) g/mL和5.0×10~(-7)~1.0×10~(-5) g/mL范围内呈良好的线性关系,检出限(3σ)分别为6.6×10~(-8) g/mL和3.7×10~(-8) g/mL。该方法已成功用于兔血浆中氢溴酸高乌甲素和富马酸异丙吡仑的同时检测,加标回收率在95.6 %~103.0 %之间。
Capillary electrophoresis(CE) with electrochemiluminescence (ECL) detection, as an high-efficient and high-sensitive analytical technique, has been extensively applied in the fields of pharmaceutical, biochemical, clinical and food analysis. But it is a pity that only a few studies on assay of environmental analytes have been reported by CE-ECL detection. On the other hand, the sensitivity and stability of a CE-ECL method will be greatly decreased since the surface of bare platinum working electrode is often contaminated by adsorbing sample matrices in biological fluids. Therefore, in this paper, a microdisk platinum electrode modified with europium (III)-doped prussian blue analogue (Eu-PB) film was used as a working electrode. Given this, a series of Ru(bpy)32+-based ECL detection coupled with CE have been established for the determination of polyoxin B absorbed in soils, ketotifen fumarate in pharmaceuticals and human urine, and lappaconitine hydrobromide, isopropiram fumarate in rabbit plasma. The present work would be helpful to the further development of detection methods for some environmental analytes and extend applications of CE-ECL technique.
This paper consists of four chapters:
Chapter One Review
The basic principles of CE and ECL are introduced firstly, and then the characteristics, the common systems and the application of CE-ECL technique in the fields of environmental and pharmaceutical analysis are summarized. Finally, the prospect of the newly developing techniques and further trends of CE-ECL methods are reviewed, and 116 references are cited.
Chapter Two Determination of polyoxin B in soils by capillary electrophoresis - electrochemiluminescence detection
On the basis of a chemically modified platinum electrode by europium (III)-doped prussian blue analogue film (Eu-PB) as a working electrode, a rapid and sensitive method for determination of polyoxin B was established using CE-ECL detection. Parameters related to the separation and detection were discussed and optimized. Under the optimized conditions, polyoxin B was separated within 4 min, and the ECL intensity was in proportion to polyoxin B concentration over the range of 1.0×10~(-7)~5.0×10~(-7) g/mL and 5.0×10~(-7)~5.0×10~(-5) g/mL with a detection limit (3σ) of 9.2×10-8 g/mL. The relative standard deviations (RSD) of intra-day and inter-day ECL intensity and migration time were less than 5.0 %. The proposed method was successfully applied in the determination of polyoxin B absorbed in three kinds of soils (e.g. bentonite, kaolinite and loess) with the recoveries of 92.2 % ~104.3 %. The results also showed that the adsorption rate of polyoxin B in soils for 2 h was 89.0 % for bentonite, 45.5 % for kaolinite and 40.4 % for loess, respectively, indicating that the adsorption capacity of polyoxin B in different soils was bentonite> kaolinite>loess. The possible reason was related to the characteristics, specific surface area and cation exchange capacity (CEC) of soils.
Chapter Three Determination of ketotifen fumarate in pharmaceuticals and human urine by capillary electrophoresis - electrochemiluminescence detection
On the basis of a chemically modified platinum electrode by europium (III)-doped prussian blue analogue film (Eu-PB) as a working electrode, a simple, rapid and sensitive CE-ECL method was developed for the determination of ketotifen fumarate (KTF). The effects of several factors such as the applied potential, the acidity and the concentration of running buffer, separation voltage, and injection conditions on the detection were investigated. Under the optimized conditions, the ECL intensity was in proportion to KTF concentration over the range of 3.0×10-8~2.0×10~(-6) g/mL and 2.0×10~(-6)~5.0×10~(-6) g/mL with a detection limit (3σ) of 2.1×10-8 g/mL. The relative standard deviations of the ECL intensity and the migration time were 0.95 and 0.26 %, respectively. The developed method was successfully applied to the determination of KTF contents in pharmaceuticals and human urine with the recoveries between 99.5 and 107.0 %.
Chapter Four Simultaneous determination of lappaconitine hydrobromide and isopropiram fumarate in rabbit plasma by capillary electrophoresis - electrochemiluminescence detection
A rapid and sensitive method for simultaneous determination of lappaconitine hydrobromide (LH) and isopropiram fumarate (IF) has been firstly established by CE - ECL detection. The effect of analytical conditions, such as applied potential, the pH and concentration of running buffer,separation voltage, injection voltage and injection time were investigated in detail. Under the optimized conditions, a baseline separation for the two analytes was achieved within 6 min, and the ECL intensity was in proportion to LH and IF concentration over the range of 1.0×10~(-7)~5.0×10~(-6) g/mL and 5.0×10~(-6)~5.0×10~(-5) g/mL for LH and 4.0×10~(-8)~5.0×10~(-7) g/mL and 5.0×10~(-7)~1.0×10~(-5) g/mL for IF with a detection limit (3σ) of 6.6×10~(-8) g/mL for LH and 3.7×10~(-8) g/mL for IF, respectively. The applicability of the proposed method was illustrated for the determination of LH and IF in rabbit plasma with recoveries between 95.6 and 103.0 %.
引文
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