【作者】 王畅；

【导师】 章竹君；

【作者基本信息】 陕西师范大学 ， 分析化学， 2002， 硕士

【摘要】 发光分析是分子发光光谱分析的简称，它主要包括分子荧光，磷光和化学发光分析（包括生物发光分析）三大部分。近几十年来，随着高灵敏度、高精度，高分辨和多功能现代发光分析仪器的引进和我国分析仪器制造业的发展，许多现代发光分析方法和技术在我国得到了发展。 由第一电子激发单重态所产生的辐射跃迁而伴随的发光称为荧光，它是一种光致发光现象。基于对化合物的荧光性质测量而建立起来的分析方法称为分子荧光光谱法。自从1867年Goppelsroder进行历史上首次的荧光分析工作后，经过一百多年的发展，除常规的方法，如通过化学反应将非荧光物质转换为适合于测定的荧光物质、荧光猝灭以及能量转移外，随着激光、计算机和电子学的新成就等一些新的科学技术的引入，同步荧光、导数荧光、时间分辨荧光、荧光偏振、荧光免疫、低温荧光、固体表面荧光等诸多新方法以及荧光反应速率法，三维荧光光谱技术和荧光光纤化学传感器等得到了很大的发展。如今，荧光分析法在生物反应器控制、生物传感器、药物监测、生化分析以及复杂体系多组分同时分析中受到人们的广泛关注并成为一种重要而有效的发光分析技术。 化学发光分析是根据化学反应产生的光辐射（化学发光）确定物质含量的一种痕量分析方法。1877年，Radziszewski发现在碱性介质中，洛酚与氧发生化学反应产生一种绿色的光，这就是首次利用合成化合物观察到的化学发光现象。此后，人们合成了一系列化学发光化合物并研究了它们的化学发光特性。在液相化学发光中常用的体系有：①咪唑类化合物（如洛酚类）；②吖啶类化合物（包括光泽精，吖啶酯）③酰肼类化合物（例如luminol体系）；④N-溴化琥珀酰亚胺作为氧化剂的发光体系；⑤强氧化剂体系，例如铈（Ⅳ）-亚硫酸盐、KmnO₄作氧化剂的发光体系；⑥电致化学发光体系；⑦过氧草酸酯体系。其中luminol体系是应用最多的化学发光体系，可用于无机物、有机物、生物体液、组织，新陈代谢物的测定。化学发光分析具有高的灵敏度(检出限可达10^-12～10^-18mol)、宽的线性范围（3～6个数量级）及相对来说比较简单便宜的仪器，设备等三个主要优点。它包括气相、火焰和液相化学发光三种类型，液相化学发光反应体系最多，应用也最广泛，气相及火焰化学发光反应体系一般用于环境污染监测。近十多年来发展起来的偶合反应的化学发光、化学发光免疫分析是对化学发光新技术的推广。由于化学发光分析所具有的优点，它在痕量分析、环境科学、生命科学及临床医学上得到愈来愈广泛的应用。 生物系统的发光现象称为生物发光。它是生物物质进行生物化学反应时伴随的发 象，例如荧光虫、茫些细菌或真匐、原生动物、，蠕虫以及甲壳动物等所发射的 光．基于生物发椰象建立起来的腑方涤脓生物发光蛐．在生物发光分析屯 萤大虫荧光素一么荧船－体第的研党和应用囊为仪．虫荧掀瞅 独特的生椰化剂，它能粹卧反应的能扮变为光．这一朋 的绷定，灵杜度可达10勺曲！，在生物医学科学、生脓学、学和农业生物学方面椭成功的应用．生掀光分析另一个应用校广的赃体系为一 一 化的发光函应－可周来删定 的朋相偶合，用于测定许多代 酋狱环撇测册肋．目前应用生物发 还不多，其浴力有待进一步挖掘．则无拼问，生物发 及 生物医学等领域尔有着理论和应用价值．＿－。。。～＿一”－ 作中 我们 究了荧光蛐、生物发踉析以硼微敝芯片相结合的化学发光分析．各项工作橱迷如下： 第一部分一潞＿川（皿）和盐酸多西时在一定铡下反应，生成的配合物可发出荧光，建立了髓水样中痕量铝的荣先分光掀法。本方法用于自来水、矿泉水中的铝的测定，结果与较为满意．－ 第二部分：将微透析采样技术与荧光分析相结合测定了6一硫鸟嗓吟 ＊h叫y闲mn民丘＊G)与牛血清白蛋白的相互作用．在NisOlixn丹丑0“侧厂吱介 中，KMnh可M6TG，其产物县有比 6－xu M轶强的荚光．利用这一现象 缘合微谩析采样投术郴研究了特44O与牛血清白蛋白的交互作用． 第三部分；利月彦环菌（一种真芮）在M在下能发光特性设计了一种狲感 ．其发光特性与外界条件有关；实验中，我们考紊了发光腑、环坟瞰，湿度、泵速和培养龈对其发光强度的影响．狮基于生物发光的M感器适合子测定高浓度氧(Q一，二叭v／v八并且无化学试剂污染，是一种绿色化学分析法． 第四邢分：－碉眩①玫芯片是当前蛐化学颅域中的一个琐热点．我们在自制的微姚孕反应器（即微瞅芯片）上首次实现了流动的偷tilllll no卜CUZ＋＄学发贩应的测定，检出限为z“X豆ov’’．rt’ L’．并且哦了血液令的皿．更多还原

【Abstract】 Luminescence analysis is the abbreviation of molecular spectrum,including molecular fluorescence, phosphorescence and chemiluminescence analysis(including bioluminescence). For the last several years, great progresses have been made in modern luminescence analysis method and technology with introduction of high sensitivity, high precision, high resolution and multifunctional modem luminescence analysis instruments and development of analytical device manufacturing industry.Fluorescence emits when the electrons transfer from the first excited single state to the base state. Since 1867,Goppelsroder has carried out the fluorescence analysis for the first time in history, in addition to the general fluorescence analysis, such as the nonfluorescence substance being transferred into fluorescence ones which can be determined, fluorescence quenching method, energy transferring method and so on, with the introduction of laser, computers and advancement of electronics , synchronous fluorimetry, derivative fluorimetry, time-resolved fluorescence, fluorescence polarization, fluorescence immunoassay, low temperature fluorescence and other methods have greatly developed, accelerating the advent of all kinds of new fluorescence instruments. Now, the fluorescence analysis has being concerned in biological reaction control, biosensor, Pharmaceuticals minitor, biochemical analysis and multi - content in complicated system. And it has been developed into an important effective spectrum analysis technology.Chemiluminescence analysis is a trace analysis method which determines the content of substance according to the light emission of the chemical reaction. In 1877, Radziszewski found that lophine emmtted green light when it reacted with oxygen in the prescence of a base. This is the first example of chemiluminescentce using a synthetic organic compound. Since then, a number of chemiluminescent compounds have been synthesized and studied for their chemiluminescent properties. Liquid chemiluminescence includes imidazole compounds( for example lophine), acridine compounds ( such as lucigengen and acridinium ester) , acidamide compounds, cerium-sulfite luminescence system, potassium permanganate, peroxyoxalate systems. In above systems, luminol system is applied widely. Luminol system can be used in inorganic substance, organic substance, Pharmaceuticals, biological body fluid, tissue and the determination of metabolism substance. Chemiluminescence analysis has high sensitivity (Detect limit canbe 10~12~ 10"18mol), wide line range( three to six magnitude order ) and simpler, lower instruments. There are gas phase, flame and liquid chemiluminescence reaction systems. Chemiluminescence method with coupling reaction, chemiluminescence immunoassay which has developed for recent ten years is the popularization of chemiluminescence technology. For the above advantages of its, Chemiluminescence has been widely applied in environment science, biological science and clinical science.The luminescence happening in biological system is called as bioluminescence which comes from the biochemical reaction of organism, such as the light of firefly, some germs or fungi, protests, worms and crustaceans. And the respound analytical method is called bioluminescence analysis. In bioluminescence analysis, luciferin - luciferase - ATP system is most widely studied and applied. Luciferase is a special biocatalyst, which can transfer the energy of enzymic reaction into light. This reaction system has been widely applied in the dermination of ATP, and the sensitivity may achieve 10~19 mol. It has been successfully applied in biomedical science, biological science, cosmic science, material medica and biology in agriculture. The other reaction system applied in bioluminescence is the luminescent reaction which is catalyzed by NAD(P)H:FMN oxydo - reductase and bacteria luciferase. It can determine NAD(P)H. Coupled with reaction of many dehydrogenase, biluminesce reaction may determine many metabolite. In addition, the papers which monitor the envir更多还原