【作者】 万芳利；

【导师】 金利通；

【作者基本信息】 华东师范大学 ， 分析化学， 2004， 硕士

【摘要】 高效液相色谱电化学检测技术自70年代诞生以来，由于其具有分离性能好、进样量少、灵敏度高等优点，已经成为化学、医学、生物、环境等领域的重要分析手段。二十一世纪是生命科学快速发展的时期，对生物样品信息的准确分析至关重要，但生物样品成分特别复杂，含量又往往很低，这就对如何实现在体、实时、连续的监测微量和超微量的生物样品提出了更高的要求。不断发展的液相色谱．电化学检测技术正好适应了这一需求，成为了检测生物样品的关键手段之一。但是，现有的技术还存在着选择性和灵敏度等不尽人意的地方，本论文以新型液相色谱电化学检测器的研究开发为重点，采用合适的生物样品处理技术与液相色谱电化学检测技术，建立了在生命体系中进行采样、分离和分析的新方法，通过研制多种修饰电极用于液相色谱电化学检测器，提高了活体分析的灵敏度和选择性。本论文以纳米材料电化学检测器研制为重点，从以下几方面对高效液相色谱电化学检测技术在生物样品分析中的应用进行了探讨： 1．TA／IrO₂-Pd修饰电极用于液相色谱电化学检测大鼠脑微渗析液中单胺类神经递质的研究与应用 首次研制了一种新型TA／IrO₂-Pd化学修饰电极用于液相色谱电化学检测。实验表明，该电极对多种单胺类神经递质及其代谢产物具有很好的催化作用，能够选择性地通过多巴胺等阳离子而排除一些脑中高浓度阴离子的干扰。将该修饰电极作为电化学检测器与高效液相色谱联用，有效地分离和检测了大鼠脑微渗析液中的去甲肾上腺素、肾上腺素、多巴胺、3，4-二羟苯乙酸和5-羟色胺等神经递质及其代谢产物。单胺类神经递质是调节机体生理活动的重要物质基础，是中枢神经系统重要的信息传递物质，由于神经递质之间的生理和药理作用密切，因此脑组织中不同神经递质的活体检测，对于脑生理和神经科学的研究具有重要的意义。 2．掺锰纳米二氧化铅修饰电极色谱电化学用于大鼠脑内的单胺类神经递质和辅酶四氢生物喋呤的应用研究中文摘要 研制了一种新型的微渗析取样一电化学检测装置，首次采用掺锰纳米二氧化铅修饰电极在微渗析取样系统中对大鼠脑内纹状体中的单胺类神经递质及代谢产物和四氢生物喋吟进行了同时检测。对大鼠腹腔注射四氢生物喋吟后，研究了大鼠脑中四氢生物喋吟浓度对单胺类神经递质及代谢产物浓度变化的影响。由于四氢生物喋吟也是一些神经疾病的临床药物，通过对实验动物的实际检测，对四氢生物喋吟产生作用的原理进行了初步的探讨，为进一步研究这一药物的疗效提供了极为有用的分析方法。3.碳纳米管修饰电极一液相色谱电化学检测用于肝细胞中还原型及氧化型谷肤甘肤的研究 还原型谷胧甘肤是体内调节氧化还原状态的重要物质，其与氧化型谷肤甘肤的比值则是反映细胞过氧化伤害程度的灵敏指标。但是，对还原型谷胧甘肤和氧化型谷肤甘肤的同时检测还缺乏一种准确、快速、灵敏的检测方法。本论文研制了新型的功能化碳纳米管修饰电极，并首次应用于还原型谷肤甘肤和氧化型谷肤甘肤的同时检测。当该修饰电极作为电化学检测器与高效液相色谱联用时，能有效地分离和检测肝细胞中的还原型和氧化型谷胧甘肤，这对医学和生物学中细胞过氧化伤害情况的分析提供了一种简便、灵敏的方法。更多还原

【Abstract】 As an important approach, high performance liquid chromatography (HPLC) -electrochemical detection (ED) has been used in the fields of chemistry, medicine, biology, environment and other sciences because of its effective separation, small injection volume and high sensitivity. In the 21st century, the research on life science is the most active. To realize the in vivo and on-line determination of biological micro-samples plays a critical role to the development of biology, medicine and to the life science, and the analysis of biological samples is the key point. HPLC-ED accommodates itself to this situation properly. However, the conventional HPLC-ED has its disadvantages, such as selectivity and sensitivity, etc. So, the development of high-quality HPLC-ED plays are of great value not only in analytical chemistry but also in biology, medicine and even in life science.In this dissertation, we laid our emphasis on the research of novel electrochemical detectors (ED), and established some new methods of sample, separation and analysis in life science by HPLC-ED coupled with samples treating measures, such as microdialysis sampling technique. Several kinds of chemically modified electrodes (CME) which could enhance the sensitivity and selectivity in the analysis of biological samples were developed and used as HPLC-ED. Nano material was applied in the HPLC-ED to enhance the sensitivity. The dissertation includes three parts, 1. In vivo determination of monoamine neurotransmitters in rat brain microdialysate by liquid chromatography with a thioctic/palladium-iridium oxide modified electrodeThe fabrication and application of a novel electrochemical detector (ED) with the thioctic/ palladium-iridium oxide modified electrode for liquid chromatography (LC) were described. The chemically modified electrode (CME) had a good catalytic effect on monoamine neurotransmitters and their metabolites. It was also found that the CME could be selective to dopamine (DA) cations and exile the ascorbic acid (AA) anions inrat brain. NE, E, DA, DOPAC, 5-HT in rat brain were also separated and determined effectively at the CME. Because the monoamine neurotransmitters are important substantial basis for physiologic activity and central nervous system, it is important to determine the monoamine neurotransmitters in vivo for cerebral physiology and neuroscience.2. Liquid chromatography with amperometric detection at a nanocrystalline Me-doped lead dioxide film modified electrode for determination of neurotransmitters and tetrahydrobioptrinSince the tetrahydrobioptrin was an important cofactor for neurosystem, a method of its sensitively detection are therefore important. In this paper, the fabrication and application of a novel electrochemical detector (ED) with nano crystalline Me-doped lead dioxide film chemically modified electrode (CME) for liquid chromatography (LC) were described. The Mn-doped PbOi film was characterized by scanning tunnel microscope (STM). It was found that the CME exhibited efficiently electrocatalytic effect on tetrahydrobioptrin and monoamine neurotransmitters. The application of this method coupled with microdialysis sampling for the determination of tetrahydrobioptrin and monoamine neurotransmitters hi rat brain was satisfactory.3. Liquid chromatography with amperometric detection using functionalized multi-wall carbon nanotubes modified electrode for simultaneous determination glutathione and glutathione disulfideGlutathione (GSH) and glutathione disulfide (GSSG) are important thiols to provide defense against oxidative stress by scavenging free radicals or causing the reduction of hydrogen peroxide. The ratio GSH/GSSG is often used as a sensitive index of oxidative stress in vivo. However, the major method of determination of them, fluorescence assay, was indirect and time consuming. In this paper, a direct electrochemical method was described by liquid chromatography (LC) coupling with electrochemical detector (ED) using an electrode modified with founctionalized carbon nan更多还原