【作者】 刘波；

【导师】 王金中；

【作者基本信息】 河南大学 ， 分析化学， 2004， 硕士

【摘要】 发光分析是分子发光光谱分析的简称，它主要包括化学发光分析（包括生物发光分析），分子荧光和磷光三大部分。 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，洛粉碱与氧发生化学反应产生一种绿色的光，这是首次利用合成化合物观察到的化学发光现象。它包括气相、火焰和液相化学发光三种类型。化学发光分析具有灵敏度高(检测限可达10^-12-10^-18mol／L)、线性范围宽（3-6个数量级）及仪器设备简单便宜等优点。由于化学发光分析具有上述的优点，它在痕量分析、环境科学、生命科学及临床医学上得到广泛应用。 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、荧光淬灭以及能量转移外，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。 本研究工作中，主要对荧光分析和化学发光分析在药物中的应用进行了研究。 各项研究工作简述如下： 一、头孢曲松钠的流动注射化学发光抑制法测定 基于吖啶橙在氢氧化钠介质中，能被高锰酸钾氧化产生较强的化学发光，头中文摘要抱曲松钠能强烈抑制其化学发光，建立了高锰酸钾一叮陡橙一头抱曲松钠化学发光抑制测定头抱曲松钠的方法;头抱曲松钠的质量浓度与化学发光强度在2一20mg/L范围内呈良好的线性关系，方法检出限为8 p g/L，对10 mg/L头抱曲松钠试液连续进行n次测定的相对标准偏差为3.1%。方法用于头抱曲松钠针剂测定，取得了满意的结果。二、流动注射化学发光法测定甲氧节氨嗜陡 本文基于荧光染料毗罗红G（Pyronine）在硫酸介质中，能被高锰酸钾氧化产生较强化学发光，甲氧节氨嚓睫加入能增强化学发光，建立了高锰酸钾一毗罗红G一甲氧节氨啼睫化学发光体系测定甲氧节氨嗜咤的新方法。甲氧节氨啼陡的浓度与化学发光强度在1.ox10一49/mL一3.ox10一，g/mL范围内呈良好的线性关系，方法的检出限为3.4x10一99/mL，对5.0x10一59/mL甲氧节氨啼陡试液进行11次测定的相对标准偏差为2.6%。该法已用于甲氧节氨喀咤片的测定，结果与药典法基本一致。三、流动注射化学发光抑制法测定毗罗昔康 本文基于叮咤橙在氢氧化钠介质中，能被高锰酸钾氧化产生较强的化学发光，毗罗昔康能强烈抑制其化学发光，建立了高锰酸钾一叮咤橙一毗罗昔康化学发光抑制测定毗罗昔康的新方法。毗罗昔康的浓度与化学发光强度在1.Ox 10一，一7.0火10一4g/mL浓度范围内呈良好的线性关系，方法检出限为4.5 x 10一69/mL，对1，ox10一5留m毛毗罗昔康试液连续进行6次测定的相对标准偏差为3.6%。四、巴比妥的流动注射化学发光法测定 本文基于过氧化氢和次氯酸钠在碱性介质中协同氧化巴比妥产生化学发光，建立了化学发光测定巴比妥的新方法。巴比妥的浓度在7.0 X10一5一5.0 X10一，孙mol/L范围内与化学发光强度呈良好的线性关系，检出限为4.6 x 10一7 mol/L。对1 .0xlo一4mol/L巴比妥试液进行6次平行测定的相对标准偏差为1.23%。五、荧光衍生法测定构椽酸喷托维林的含量 采用衍生化反应，建立了荧光分光光度法测定构椽酸喷托维林的新方法，并 河南大学分析化学专业20川级硕士学位论文 刘 波 对影响荧光强度的衍生体系、反应温度、反应时间、产物稳定性等进行了研究。 实验表明在激发波长和发射波长分别为 367 urn和 457nLm下测得荧光强度与其浓度 在 8刀 XIO”’～1．0 XIO“g／mL范围内呈良好的线性关系，并对 1刀 XIO”旮mL拘激 酸喷托维林溶液进行6次平行测定的相对标准偏差为0．31％，检出限达15X 10”为mL。本法己用于拘檬酸喷托维林片剂含量的测定，结果与药典法测得值相符。 六、流动注射荧光法测定替硝哩 采用铁一冰醋酸还原体系，将替硝哩分子中的硝基还原为氨基后，测其含量， 据此建立了替硝哇的荧光分析法，在激发波长360urn，发射波长420urn处测定其 荧光强度，替硝哇浓度在 4．0 XIO”‘mol几～4．0 XIO”‘mol几范围内与其荧光强度呈 良好的线性关系，本法己用于替硝硅片剂和替硝峻葡萄糖注射液测定，结果与药 典法基本一致。 七、流动注射荧光法测定奥硝哩 采用铁一冰醋酸还原体系，将奥硝哩分子中的硝基还原更多还原

【Abstract】 Luminescence analysis is the abbreviation of molecular spectrum, including molecuar fluorecence phosphorescence and chemliluminescence analysis (including bioluminescence) .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 emmitted green light when it reacted with oxygen in the prescence of a base.This is the first example of chemiluminescence using a synthetic organic compound. There are gas phase, flame and liquid chemiluminescence reaction systems, chemiluminescence analysis can has high sensitivity (Detect limit can be 10-12-10-18mol/L), wide linear range (three to six magnitude order) and simpler, lower instruments. Chemiluminescence method with coupling reaction, chemiluminescence immunoassay which has developed for recent ten years is the popularization of chemiluminescence technology. For the above advantages, Chemiluminescence has been widely applied in environment science, biological science and clinical science.Fluorescence emits when the electrons transfer from the fisrt 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, synchromous fluorimetry , deivative fluorimetry , time-resolved fluorescence , fluorescence polaruzation, 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 inbiological 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. The thesis consists of six parts:Part one Determination of Ceftriaxone Sodium by Quenchingof Flow Injection ChemiluminescenceIntensity chemiluminescence can be emitted by oxidation reaction of potassium permanganate with acridine orange in sodium hydrogen medium, chemiluminescence is greatly quenched by the addition of Ceftriaxone Sodium .The calibration curve is linear over the range of 2~20mg/L.The detection limit is 8ng/mL. A 3.1% relative standard deviation is obtained in a repeatability study (n=11) at the 10mg/L level of ceftriaxone sodium. The method has been applied to determine ceftriaxone sodium in pharmaceutical with good result.Part two Study on Chemiluminesecence Reaction of Trimethoprim- KMnO4- Pyronine -H2SO4 by Flow Injection MethodA new CL reaction of Trimethoprim - KMn(V Pyronine -H2SO4 was studied by flow injection method.The factors of influencing CL intensity were investigated .A new method based on the CL reaction for the determination of TMP has been established by optimizing the experimental conditions.The linear range for the determination of TMP was 1.0 X10-4 mol/L~3.0 X 10-7mol/L and the determination limit was 3.4 X 10-9mol/L .The relative standard for the determination of 5.0 X 10-5mol/L TMP (n = 11) was 2.6% .The method has been used to determination in tablets and the results were in good agreement with those obtained by classical method .Part three Determination of Piroxicam by Quenchingof The Chemiluminescence A method is described for the determination of Piroxicam by quenching of thechemiluminescence signal of the acridine orange-KMnO4-OH-. The method is based on the fact that the concentration of potassium permanganate solution is lower by the reaction of permanganate with Piroxicam.The method is sensitive, the limit of detection is 4.5 X 10-6g/mL. The linear range of the working curves is 1.0X10-5~7.0X 10-4 g/mL. The relative standard devia更多还原