基于吖啶酯化学发光体系的毛细管电泳—化学发光在线联用技术在药物分析中的应用
发布时间：2018-07-28 11:35
【摘要】：随着医疗事业的不断发展,卫生医疗条件的不断改善,人们在日常用药中也更加注重安全有效。因此,治疗药物浓度的监测工作在保障人们用药安全中也显得越来越重要。由于生物样品种类繁多,成分复杂等特点,对治疗药物监测手段的发展、更新不断提出新的挑战。因此,建立使用方便快捷、操作简单、灵敏度高、选择性强的分析方法是目前体内药物分析研究的重要内容。毛细管电泳(capillary elelctrophoresis, CE)分离技术具有分离时间短、分离效率高、样品消耗少等优点,已成为近年来药物分析最活跃的研究方向之一。为适应分析技术的发展要求,CE已经开拓了不同的分离模式,从而能满足对不同性质、种类样品的分析检测要求,这也使得其在检测生物体液中药物及药物代谢物等方面得到了广泛应用。但由于CE分析量都在极微量水平,这就决定了其搭载的检测技术必须具备足够的灵敏度才能达到生物样品的检测要求。虽然激光诱导荧光检测器(laser induced fluorescence detector, LIF)、质谱检测器(mass spectrometry detector, MS)等检测手段具备极高的检测灵敏度,但其昂贵的仪器成本、复杂的检测程序极大的限制了它们的应用。化学发光(chemiluminescence, CL)检测技术是在暗背景条件下对光信号搜集检测,是一种在不需要外加光源的情况下即可实现检测的分析手段,不仅仪器简单廉价、易操作,而且能有效避免了光源的不稳定以及瑞利散射和拉曼散射引起的背景噪音,使得其具有极高的检测灵敏度,在一定程度上甚至可以与MS、LIF等检测技术相媲美。因此,CE与CL技术的结合是一种极为理想的分离分析手段,在医药分析领域,特别是在药代动力学及相关领域具有极高的应用潜力。本论文主要基于CE和CL各自的优点,以吖啶酯作为柱前CL衍生试剂,建立了CE-CL的在线联用技术用于同时检测多巴胺(dopamine, DA)和西布特罗(cimbuterol, CM)。具体研究工作简述如下： 以吖啶酯衍生物2’,6'-二甲基-4’-(N-琥珀酰亚胺基)苯基-10-甲基吖啶-9-羧酸-1-丙磺酸内(2',6'-dimethyl-4'-(N-succinimidyloxycarbonyl)phenyl-10-methyl-acridinium-9-carboxylate-1-prop anesulfonate inner salt, AE)作为柱前CL衍生试剂,建立了一种简单快速的CE-CL方法,实现了DA及CM的同时高灵敏度检测。实验中详细考察了影响毛细管分离效率、CL信号强度的因素以及氧化剂、共反应剂混合模式对系统分析性能的影响。在最优条件下,DA与CM的浓度分别在50-1500ng/mL及2.0-1000ng/mL范围内与增强的CL强度呈良好的线性关系,检测限(S/N=3)分别为2.0ng/mL和0.50ng/mL o用本方法对人尿液中的DA进行了检测,并以酶联免疫分析(enzyme linked immunosorbent assay, ELISA)试剂盒法作为参照实验,两种方法所得结果一致。尿液中DA的加标回收率在83.7%至90.5%之间,展现了良好的应用潜力。
[Abstract]:With the development of medical service and the improvement of medical condition, people pay more attention to safety and efficiency in daily medication. Therefore, the monitoring of therapeutic drug concentration is becoming more and more important in ensuring the safety of drug use. Because of the variety of biological samples and the complexity of the components, the development of therapeutic drug monitoring methods and the renewal of new challenges are constantly put forward. Therefore, the establishment of a convenient, simple, sensitive and selective analytical method is an important content of drug analysis in vivo. Capillary electrophoresis (CE) (capillary elelctrophoresis, CE) separation technology has been one of the most active research directions for drug analysis in recent years because of its advantages of short separation time, high separation efficiency and low sample consumption. In order to meet the requirements of the development of analytical technology, CE has developed different separation modes, so that it can meet the requirements for the analysis and detection of samples of different properties and types. It has also been widely used in the detection of drugs and drug metabolites in biological fluids. However, due to the very small amount of CE analysis, it is decided that the detection technology must have sufficient sensitivity to meet the detection requirements of biological samples. Although the laser induced fluorescence detector (LIF) (laser induced fluorescence detector, LIF), mass spectrometer (mass spectrometry detector, MS) has very high detection sensitivity, its expensive instrument cost and complex detection program greatly limit their application. Chemiluminescence (chemiluminescence, CL) detection technology is a kind of analytical means to detect light signal under dark background, which can be detected without additional light source. It is not only simple and cheap, but also easy to operate. Moreover, it can effectively avoid the instability of light source and background noise caused by Rayleigh scattering and Raman scattering, which makes it have very high detection sensitivity, and to some extent it can be compared with other detection techniques such as MSLIF. Therefore, the combination of CE and CL technology is an ideal method of separation and analysis, and has a high application potential in the field of pharmaceutical analysis, especially in pharmacokinetics and related fields. In this paper, based on the advantages of CE and CL, an on-line combination technique of CE-CL was established for the simultaneous detection of dopamine (dopamine, DA) and sibuterol (cimbuterol, CM). With acridine ester as precolumn CL derivative reagent. The specific research work is summarized as follows: the acridine ester derivative 2Acridine 6-dimethyl-4- (N-succinylimido) phenyl -10-methylacridine-9-carboxylic acid -1-propanesulfonic acid (N-succinimidyloxycarbonyl) phenyl-10-methyl-acridinium-9-carboxylate-1-prop anesulfonate inner salt, AE) was used as pre-column CL derivatization reagent. A simple and fast CE-CL method is established to detect DA and CM with high sensitivity. The factors which affect the capillary separation efficiency and the influence of the mixture of oxidant and co-reaction agent on the performance of the system were investigated in detail. Under the optimal conditions, the concentration of DA and CM in the range of 50-1500ng/mL and 2.0-1000ng/mL showed a good linear relationship with the enhanced CL strength. The detection limit (S/N=3) was 2.0ng/mL and 0.50ng/mL o respectively. The method was used to detect DA in human urine. The Elisa (enzyme linked immunosorbent assay, ELISA) kit was used as the reference test. The results of the two methods were in agreement with each other. The recoveries of DA in urine ranged from 83.7% to 90.5%, showing good application potential.
【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R914;R917
本文编号：2150006
[Abstract]:With the development of medical service and the improvement of medical condition, people pay more attention to safety and efficiency in daily medication. Therefore, the monitoring of therapeutic drug concentration is becoming more and more important in ensuring the safety of drug use. Because of the variety of biological samples and the complexity of the components, the development of therapeutic drug monitoring methods and the renewal of new challenges are constantly put forward. Therefore, the establishment of a convenient, simple, sensitive and selective analytical method is an important content of drug analysis in vivo. Capillary electrophoresis (CE) (capillary elelctrophoresis, CE) separation technology has been one of the most active research directions for drug analysis in recent years because of its advantages of short separation time, high separation efficiency and low sample consumption. In order to meet the requirements of the development of analytical technology, CE has developed different separation modes, so that it can meet the requirements for the analysis and detection of samples of different properties and types. It has also been widely used in the detection of drugs and drug metabolites in biological fluids. However, due to the very small amount of CE analysis, it is decided that the detection technology must have sufficient sensitivity to meet the detection requirements of biological samples. Although the laser induced fluorescence detector (LIF) (laser induced fluorescence detector, LIF), mass spectrometer (mass spectrometry detector, MS) has very high detection sensitivity, its expensive instrument cost and complex detection program greatly limit their application. Chemiluminescence (chemiluminescence, CL) detection technology is a kind of analytical means to detect light signal under dark background, which can be detected without additional light source. It is not only simple and cheap, but also easy to operate. Moreover, it can effectively avoid the instability of light source and background noise caused by Rayleigh scattering and Raman scattering, which makes it have very high detection sensitivity, and to some extent it can be compared with other detection techniques such as MSLIF. Therefore, the combination of CE and CL technology is an ideal method of separation and analysis, and has a high application potential in the field of pharmaceutical analysis, especially in pharmacokinetics and related fields. In this paper, based on the advantages of CE and CL, an on-line combination technique of CE-CL was established for the simultaneous detection of dopamine (dopamine, DA) and sibuterol (cimbuterol, CM). With acridine ester as precolumn CL derivative reagent. The specific research work is summarized as follows: the acridine ester derivative 2Acridine 6-dimethyl-4- (N-succinylimido) phenyl -10-methylacridine-9-carboxylic acid -1-propanesulfonic acid (N-succinimidyloxycarbonyl) phenyl-10-methyl-acridinium-9-carboxylate-1-prop anesulfonate inner salt, AE) was used as pre-column CL derivatization reagent. A simple and fast CE-CL method is established to detect DA and CM with high sensitivity. The factors which affect the capillary separation efficiency and the influence of the mixture of oxidant and co-reaction agent on the performance of the system were investigated in detail. Under the optimal conditions, the concentration of DA and CM in the range of 50-1500ng/mL and 2.0-1000ng/mL showed a good linear relationship with the enhanced CL strength. The detection limit (S/N=3) was 2.0ng/mL and 0.50ng/mL o respectively. The method was used to detect DA in human urine. The Elisa (enzyme linked immunosorbent assay, ELISA) kit was used as the reference test. The results of the two methods were in agreement with each other. The recoveries of DA in urine ranged from 83.7% to 90.5%, showing good application potential.
【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R914;R917
【参考文献】
相关期刊论文 前2条
1 赵鹏程;王彦吉;;光二极管阵列检测毛细管电泳法分析滚珠水性笔墨水[J];光谱学与光谱分析;2009年04期
2 赵利霞,林金明,屈锋;微板式磁化学发光酶免疫分析法对人绒毛膜促性腺激素(HCG)的灵敏快速测定[J];化学学报;2004年01期
，本文编号：2150006
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