基于吖啶酯化學(xué)發(fā)光體系的毛細(xì)管電泳—化學(xué)發(fā)光在線聯(lián)用技術(shù)在藥物分析中的應(yīng)用
發(fā)布時(shí)間：2018-07-28 11:35
【摘要】：隨著醫(yī)療事業(yè)的不斷發(fā)展,衛(wèi)生醫(yī)療條件的不斷改善,人們在日常用藥中也更加注重安全有效。因此,治療藥物濃度的監(jiān)測工作在保障人們用藥安全中也顯得越來越重要。由于生物樣品種類繁多,成分復(fù)雜等特點(diǎn),對治療藥物監(jiān)測手段的發(fā)展、更新不斷提出新的挑戰(zhàn)。因此,建立使用方便快捷、操作簡單、靈敏度高、選擇性強(qiáng)的分析方法是目前體內(nèi)藥物分析研究的重要內(nèi)容。毛細(xì)管電泳(capillary elelctrophoresis, CE)分離技術(shù)具有分離時(shí)間短、分離效率高、樣品消耗少等優(yōu)點(diǎn),已成為近年來藥物分析最活躍的研究方向之一。為適應(yīng)分析技術(shù)的發(fā)展要求,CE已經(jīng)開拓了不同的分離模式,從而能滿足對不同性質(zhì)、種類樣品的分析檢測要求,這也使得其在檢測生物體液中藥物及藥物代謝物等方面得到了廣泛應(yīng)用。但由于CE分析量都在極微量水平,這就決定了其搭載的檢測技術(shù)必須具備足夠的靈敏度才能達(dá)到生物樣品的檢測要求。雖然激光誘導(dǎo)熒光檢測器(laser induced fluorescence detector, LIF)、質(zhì)譜檢測器(mass spectrometry detector, MS)等檢測手段具備極高的檢測靈敏度,但其昂貴的儀器成本、復(fù)雜的檢測程序極大的限制了它們的應(yīng)用�；瘜W(xué)發(fā)光(chemiluminescence, CL)檢測技術(shù)是在暗背景條件下對光信號搜集檢測,是一種在不需要外加光源的情況下即可實(shí)現(xiàn)檢測的分析手段,不僅儀器簡單廉價(jià)、易操作,而且能有效避免了光源的不穩(wěn)定以及瑞利散射和拉曼散射引起的背景噪音,使得其具有極高的檢測靈敏度,在一定程度上甚至可以與MS、LIF等檢測技術(shù)相媲美。因此,CE與CL技術(shù)的結(jié)合是一種極為理想的分離分析手段,在醫(yī)藥分析領(lǐng)域,特別是在藥代動(dòng)力學(xué)及相關(guān)領(lǐng)域具有極高的應(yīng)用潛力。本論文主要基于CE和CL各自的優(yōu)點(diǎn),以吖啶酯作為柱前CL衍生試劑,建立了CE-CL的在線聯(lián)用技術(shù)用于同時(shí)檢測多巴胺(dopamine, DA)和西布特羅(cimbuterol, CM)。具體研究工作簡述如下： 以吖啶酯衍生物2’,6'-二甲基-4’-(N-琥珀酰亞胺基)苯基-10-甲基吖啶-9-羧酸-1-丙磺酸內(nèi)(2',6'-dimethyl-4'-(N-succinimidyloxycarbonyl)phenyl-10-methyl-acridinium-9-carboxylate-1-prop anesulfonate inner salt, AE)作為柱前CL衍生試劑,建立了一種簡單快速的CE-CL方法,實(shí)現(xiàn)了DA及CM的同時(shí)高靈敏度檢測。實(shí)驗(yàn)中詳細(xì)考察了影響毛細(xì)管分離效率、CL信號強(qiáng)度的因素以及氧化劑、共反應(yīng)劑混合模式對系統(tǒng)分析性能的影響。在最優(yōu)條件下,DA與CM的濃度分別在50-1500ng/mL及2.0-1000ng/mL范圍內(nèi)與增強(qiáng)的CL強(qiáng)度呈良好的線性關(guān)系,檢測限(S/N=3)分別為2.0ng/mL和0.50ng/mL o用本方法對人尿液中的DA進(jìn)行了檢測,并以酶聯(lián)免疫分析(enzyme linked immunosorbent assay, ELISA)試劑盒法作為參照實(shí)驗(yàn),兩種方法所得結(jié)果一致。尿液中DA的加標(biāo)回收率在83.7%至90.5%之間,展現(xiàn)了良好的應(yīng)用潛力。
[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.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R914;R917
本文編號：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.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R914;R917
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 趙鵬程;王彥吉;;光二極管陣列檢測毛細(xì)管電泳法分析滾珠水性筆墨水[J];光譜學(xué)與光譜分析;2009年04期
2 趙利霞,林金明,屈鋒;微板式磁化學(xué)發(fā)光酶免疫分析法對人絨毛膜促性腺激素(HCG)的靈敏快速測定[J];化學(xué)學(xué)報(bào);2004年01期
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