分子印迹—化学发光法在毒物分析中的研究与应用

【作者】 刘梅；

【导师】 吕九如；

【作者基本信息】 陕西师范大学 ， 分析化学， 2004， 硕士

【摘要】 本论文由综述、研究报告两部分组成。第一部分为综述，第二部分为研究报告。研究报告由三部分组成。 第一部分简单概述了分子印迹技术的基本原理及印迹聚合物的制备方法等，着重介绍了该技术在分析化学领域中的应用。 第二部分报告了分子印迹-化学发光法在毒物分析中的具体研究工作。分子印迹技术与以强氧化剂高锰酸钾为发光试剂的化学发光体系相结合，将分子印迹-化学发光法应用于复杂样品中分析对象的研究。 化学发光分析法，以其灵敏度高、线性范围宽、仪器设备简单、分析速度快等优点被广泛应用于分析领域。但由于化学发光法的选择性差，限制了其在复杂样品分析中的应用。目前解决问题的途径多是将此法与高效液相色谱、气相色谱、毛细管电泳等分离技术相结合，但终因这些方法所需仪器昂贵，操作复杂而未能很好的推广应用。 分子印迹技术是近年来迅速发展起来的一种制备对模板分子具有特效识别能力的聚合物的技术。分子印迹聚合物(MIP)对目标分子特殊的识别能力可以和抗体-抗原、酶-底物、受体-激素等天然分子识别系统相媲美。本论文将分子印迹技术与化学发光法相结合，建立了分子印迹-化学发光分析法。高锰酸钾化学发光体系由于其成本低，适用分析对象广等特点，常应用于毒物及药物分析中。本研究工作中选择还原性物质作为MIP的保护剂，避免了强氧化剂酸性高锰酸钾对含有不饱和键的MIP的破坏作用，保持了MIP的稳定性。使得分子印迹-化学发光分析法能够应用于以强氧化剂为发光试剂的化学发光体系中，利用MIP对目标分子特异的识别和捕获能力，达到提高化学发光分析选择性的目的。分子印迹-化学发光法在毒物分析中的具体研究工作包括以下三部分： 1．分子印迹-化学发光法测定吗啡 该项研究工作的目的在于将分子印迹技术和化学发光分析法相结合，建立可以对复杂样品中吗啡含量进行直接测定的分子印迹-化学发光分析法，使得化学发光分析法既可以发挥其灵敏度高、线性范围宽、仪器设备简单等优点，又解决了其选择性差的问题。本文以甲基丙烯酸为功能单体，乙二醇二甲基丙烯酸酯为交联剂合成了对吗啡有高度选择性的MIP，将此MIP填充到化学发光分析系统的流通池中，利用MIP对吗啡特有的识别和捕获能力，使其吸附在MIP_匕而与样品中的共存组分分离，然后进行化学发光检测，提高了化学发光分析方法的选择性。在选择的最佳条件下，化学发光强度与吗啡浓度在5.ox10一9一1.0又10一6g/mL范围内具有良好的线性关系。相对标准偏差为2.8o/o(cs一1 .0 xlo一诊mL，n一9)，方法的检出限为2 x 10一gg/mL。本研究工作较为成功地对吸毒人员尿液中吗啡的含量进行了直接测定。2.马钱子碱分子印迹一化学发光传感器的研究 该项研究工作以甲基丙烯酸为功能单体，乙二醇二甲基丙烯酸酷为交联剂，利用分子印迹技术首次合成了对马钱子碱具有高度选择性的MIP。将此MIP作为分子识别和传感物质，利用高锰酸钾一马钱子碱化学发光体系，研制出了一种新型测定马钱子碱的分子印迹一化学发光传感器，并将其应用于复杂样品中马钱子碱的直接测定。此传感器对马钱子碱具有良好的选择性，评价测定马钱子碱的线性范围为5.0 xlo一9一1.0 x10一69/mL，检出限为3 xlo一gg/mL，对1.0、10一诊mL马钱子碱标准溶液进行9次平行测定的RsD为2.6%。3.甲氧节陡分子印迹一化学发光法的研究 该项研究工作以甲氧节咤为模板分子，甲基丙烯酸为功能单体，乙二醇二甲基丙烯酸酷为交联剂合成了对甲氧节陡具有高选择性的MIP。以NaZSZO3为分子印迹聚合物的保护剂，利用高锰酸钾一甲氧节睫化学发光体系建立了可以直接应用于复杂样品中甲氧节咙含量测定的分子印迹一化学发光分析法。化学发光强度与甲氧节咤浓度在l.oxlo一8一1.0xlo一6g/mL范围内具有良好的线性关系。相对标准偏差为4.1%(Cs一1.ox10一79/mL，n一9)，方法的检出限为3x10一”g/mL。本研究工作分别对复方制剂及服药后人体代谢液中甲氧节咤的含量进行了直接测定，结果较为满意。更多还原

【Abstract】 This thesis includes a review and a research section. The principle of molecular imprinting technique, the method of molecularly imprinted polymer (MIP) synthesis and the applications of this technique in analytical chemistry were described in the review. The research section is focused on the study of molecular imprinting-chemiluminescence analysis in toxin.Chemiluminescence (CL) method has been widely used in the many fields due to its inherent high sensitivity and simple instrumentation. However, the relatively poor selectivity of the CL method itself limits its direct application to the analysis of analyte in complicated sample. The common way to solve the problem is that the CL method was applied in combination with some separation means such as high-performance liquid chromatography and capillary electrophoresis, whereas, these methods required relatively complicated and expensive instruments.Molecular imprinting is a technique used for preparing polymers with synthetic recognition sites having a predetermined selectivity for analyte The excellent recognition ability to the target molecule of MIP can compare with antibody-antigen, enzyme-substrate natural molecular recognition system. The imprinting-chemiluminescence method was established with the molecular imprinting technique combined with chemiluminescence method, and it is possible to be a selective and sensitive method for the determination of analyte in complicated samples directly. 1. Molecular imprinting-chemiluminescence determination of morphine using morphine -imprinted polymer as recognition materialThe aim of this work is to establish the molecular imprinting-chemiluminescence determination of morphine by the combing of molecular imprinting technique and chemiluminescence method, it is a new method that not only can employ the advantage of chemiluminescence method but also can improve the selectivity of that method. The MIP for morphine was prepared using methacrylic acid as functional monomer and ethylene glycol dimethacrylate as cross-linker in the presence of template molecule of morphine. The MIP with special recognition ability to morphine was packed into a flow cell in the flow analysis and was applied to recognize the target molecule, afterward, the chemiluminescence reagent flowedthrough the flow cell and CL. Interference experiments demonstrated that the application of MIP to CL analysis could improve the selectivity of CL method greatly. Under the selected experimental conditions, the CL intensity responded linearly with the concentration of morphine in the range of 5.0 10-9~1.0xlO~6g/mL with the detection limit of morphine was 2 10-9g/mL. The standard deviations for 1.0 10-7g/mL of morphine solution was 2.8% (n=9). This method has been applied to the determination of morphine in the urine of the drugger.2. Molecular imprinting-chemiluminescence sensor for brucine using brucine-imprinted polymer as the recognition elementThis work developed a new molecular imprinting-chemiluminescence sensor for determination of brucine. The MIP for brucine was prepared by copolymerization of methacrylic acid (MAA) and ethylene glycol dimethacrylate (EGDMA) in the presence of brucine. The polymer was employed as the recognition element of the sensor for brucine. The CL intensity responded linearly with the concentration of brucine in the range of 5.0xlO~9~1.0xlO~6g/mL, The standard deviations for 1.0xlO~7g/mL of brucine solution was 2.6% (n=9). The detection limit was 3xlO"9g/mL. The sensor has been used in the chemiluminescent system of potassium permanganate to determination of brucine in the diluted urine samples. 3.Molecular imprinting-chemiluminescence determination of TrimethoprimThis work established a new method to determination trimethoprim of using trimethoprim -imprinted polymer as recognition material. The MIP for trimethoprim was prepared using methacrylic acid as functional monomer and ethylene glycol dimethacrylate as cross-linker in the presence of template molecule of trimethoprim. The selectivity of chemiluminescence met更多还原