荧光探针的合成及自由基检测研究
荧光探针的合成及自由基检测研究
摘要
荧光分析法在生物化学、医学、工业和化学研究中的应用与日俱增，其原因在于荧光分析法具有高灵敏度的优点，且荧光现象具有有利的时间表度。由于物质分子结构不同，其所吸收光的波长和发射的荧光波长也不同，利用这一特性可以定性鉴别物质。荧光探针技术是一种利用探针化合物的光物理和光化学特性，在分子水平上研究某些体系的物理、化学过程和检测某种特殊环境材料的结构及物理性质的方法。该技术不仅可用于对某些体系的稳态性质进行研究，而且还可对某些体系的快速动态过程如对某种新物种的产生和衰变等进行监测。这种技术具备极高的灵敏性和极宽的动态时间响应范围的基本特点。羟基自由基(HO·)和超氧阴离子自由基(O2-·)是生物体内活性氧代谢产生的物质，当体内蓄积过量自由基时，它能损伤细胞，进而引起慢性疾病及衰老效应。因此，近些年来人们为了预防这类疾病的发生，自由基的研究已逐渐成为热点。而快速、灵敏和实用的自由基检测方法就显得十分重要。荧光探针检测自由基具有操作简便、响应迅速、选择性高等多种优点，我们将着重研究一类苯并噻唑结构荧光探针的合成及其对超氧阴离子自由基(O2-·)的检测。
关键词：荧光探针，苯并噻唑，超氧阴离子自由基，自由基检测
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荧光探针的合成及自由基检测研究
SYNTHESIS OF FLUORESCENT PROBES AND
DETECTION OF FREE RADICALS
ABSTRACT
Applications of fluorescence analysis method in biochemistry, medicine, industry and chemical research grow with each passing day, the reason is that fluorescence analysis method has the advantages of high sensitivity, and the flurescence phenomenon has a favorable time characterization. Since the molecular structure of different materials, the absorption wavelength and fluorescence wavelength of the emitted light is different, this feature can be characterized using differential substances. Fluorescent probe technology is a method using photophysical and photochemical properties for researching some systems’ physical and chemical process at the molecular level and detecting a particular structure and physical property of the special environment material. This technology not only can be used for steady-state nature of certain system, but also can monitore fast dynamic processes of a certain system such as the production and decay of a new species. This technology has the basic characteristics of a high degree of sensitivity and very wide dynamic range response time. Hydroxyl radical(HO-·)and superoxide anion radical(O2-·) is a substance produced in vivo metabolism of reactive oxygen species. When the body accumulates excess free radicals that will damage cells thereby causing chronic diseases and aging effects. Thus, in recent years people in order to prevent the occurrence of such diseases, the study of free radicals has become a hot spot. And fast, sensitive and practical method for the detection is very important. Using the fluorescent probes for the detection of free radicals is a simple, quick response, high selectivity variety of advantages. We will focus on the study of a class
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荧光探针的合成及自由基检测研究
of synthetic fluorescent probes of benzothiazole structure and detection of superoxide anion radical.
Key words：Fluorescent probes, Benzothiazole, Superoxide anion radical, Detection of free radicals
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荧光探针的合成及自由基检测研究
目 录
1 绪论................................................................................................................... 1
1.1 引言 .............................................................................................................................. 1 1.2 荧光 .............................................................................................................................. 1
1.2.1 荧光的产生 ....................................................................................................... 1 1.2.2 荧光探针结构特点 ........................................................................................... 2 1.2.3 荧光探针传感机理 ........................................................................................... 3 1.2.4 常见荧光团 ....................................................................................................... 3 1.2.5 荧光探针的性能 ............................................................................................... 5 1.2.6 影响荧光探针性能的因素 ............................................................................... 5 1.2.7 荧光淬灭 ........................................................................................................... 5 1.3 自由基 .......................................................................................................................... 6
1.3.1 自由基的间接检测技术 ................................................................................... 6 1.3.2 自由基的直接检测技术 ................................................................................... 7 1.4 研究现状 ...................................................................................................................... 8
1.4.1 超氧化物歧化酶(SOD)的检测 ........................................................................ 8 1.4.2 2-(2-吡啶)-苯并噻唑啉荧光探针 ..................................................................... 8 1.4.3 PF-1和PNF-1 ................................................................................................... 8 1.4.4 香草醛缩苯胺 ................................................................................................... 8 1.4.5 Hydroethidine类荧光探针 ................................................................................ 9 1.4.6 二(2,4-二硝基苯磺酰基)二氟荧光素 .............................................................. 9 1.5 选题背景和意义 ........................................................................................................ 10 1.6 课题研究内容 ............................................................................................................ 10
2 荧光探针的合成............................................................................................. 11
2.1 引言 ............................................................................................................................ 11 2.2 还原文献 .................................................................................................................... 11
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荧光探针的合成及自由基检测研究
2.3 新探针合成 ................................................................................................................ 11
2.3.1 2-(4-二甲氨基苯)-苯并噻唑 ........................................................................... 11 2.3.2 2-(4-氰基苯)-苯并噻唑 ................................................................................... 12 2.3.3 2-(苯)-苯并噻唑 .............................................................................................. 12 2.3.4 2-(4-甲基苯)-苯并噻唑 ................................................................................... 12 2.3.5 2-(4-硝基苯)-苯并噻唑 ................................................................................... 13 2.3.6 2-(水杨醛)-苯并噻唑 ...................................................................................... 13 2.4 合成小结 .................................................................................................................... 14 2.5 实验药品及规格 ........................................................................................................ 14 2.6 实验仪器及型号 ........................................................................................................ 15
3 实验结果与讨论............................................................................................. 16
3.1 引言 ............................................................................................................................ 16 3.2 荧光性能测试 ............................................................................................................ 16
3.2.1 荧光性能待测溶液配制 ................................................................................. 16 3.2.2 荧光性能测试结果 ......................................................................................... 16 3.2.3 测试谱图 ......................................................................................................... 17 3.3 1H NMR数据 ............................................................................................................. 21
3.3.1 2-(2-吡啶)-苯并噻唑 ....................................................................................... 21 3.3.2 2-(4-二甲氨基苯)-苯并噻唑 ........................................................................... 22 3.3.3 2-(4-氰基苯)-苯并噻唑 ................................................................................... 23 3.3.4 2-(苯)-苯并噻唑 .............................................................................................. 24 3.3.5 2-(4-甲基苯)-苯并噻唑 ................................................................................... 25 3.3.6 2-(水杨醛)-苯并噻唑 ...................................................................................... 25 3.3.7 2-(2-噻吩)-苯并噻唑 ....................................................................................... 26 3.4 反应条件控制及处理 ................................................................................................ 27 3.5 结论与展望 ................................................................................................................ 27
参考文献............................................................................................................... 28 致谢....................................................................................................................... 30 译文及原文........................................................................................................... 31
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