罗丹明B类和萘酰亚胺类荧光探针的合成及性能研究
发布时间：2018-08-16 12:33
【摘要】：荧光分析方法具有灵敏度高、选择性好、分析速度快、操作简单和易于细胞原位成像等优点,该分析方法常见于生物化学、环境科学和医疗诊断等领域。同时,基于罗丹明B类和萘酰亚胺类荧光染料具有较高的荧光量子产率、良好的光热稳定性、结构容易修饰和价格低廉等性质,这两类荧光染料被普及应用。因此,本论文利用罗丹明B和萘酰亚胺为荧光母体设计合成了两种荧光探针分别应用于检测线粒体中的ATP和溶酶体中的H_2S。具体内容如下:1.设计并合成了荧光探针分子Mito-Rh和Lyso-Nh,并通过了飞行时间质谱,核磁共振氢谱和核磁共振碳谱等表征手段对其结构确认。2.我们利用罗丹明B,二乙烯三胺和三苯基膦分别作为荧光探针分子Mito-Rh的荧光团,识别基团和定位基团。荧光探针分子Mito-Rh自身无色无荧光,而当加入ATP后,ATP能与该探针分子Mito-Rh发生相互作用,使荧光团罗丹明B内酰胺环被打开,导致探针分子Mito-Rh在583 nm处发出强烈的红色荧光。该探针分子Mito-Rh对ATP具有较高的灵敏性,反应前后荧光增强了81倍,检测范围为0.1-10 mM,与线粒体中的ATP含量相匹配。同时,相比于其他生物性的阴离子,比如:ADP,AMP,GTP,CTP和UTP等,该探针对ATP具有特异的选择性,主要归功于该探针对ATP具有双识别位点—氢键相互作用和π-π相互堆积作用。此外,该探针能精准地定位于线粒体中并成功应用于实时监控线粒体中ATP的含量变化。3.我们利用萘酰亚胺,硝基苯并二唑和吗啉分别作为荧光探针分子Lyso-Nh的荧光团,识别基团和定位基团。该探针自身存在吗啉基团N原子的孤对电子和淬灭基团(识别基团)产生的PET过程影响,该探针无荧光发射,在pH=5的条件下加入H_2S时,识别基团离去且N原子的孤对电子质子化,在536 nm处产生绿色的萘酰亚胺特征荧光发射峰。反应前后荧光增强倍数为72倍,表明该探针的灵敏性较高。同时,该探针具有较好的选择性,不受其他生物性相关的物质干扰影响。此外,该探针可以检测溶酶体中的H_2S含量的变化。
[Abstract]:Fluorescence analysis method has the advantages of high sensitivity, good selectivity, fast analysis speed, simple operation and easy in situ imaging of cells. It is widely used in the fields of biochemistry, environmental science and medical diagnosis. At the same time, Rhodamine B and naphthalimide fluorescent dyes are widely used because of their high fluorescence quantum yield, good photothermal stability, easy structure modification and low price. Therefore, in this thesis, Rhodamine B and naphthalimide were used to design and synthesize two fluorescent probes for the detection of ATP in mitochondria and H _ 2s in lysosome, respectively. The details are as follows: 1. The fluorescent probe molecules Mito-Rh and Lyso-Nhwere designed and synthesized, and their structures were confirmed by time-of-flight mass spectrometry (TF-MS), nuclear magnetic resonance (NMR) and nuclear magnetic resonance (NMR) carbon spectroscopy. Rhodamine B, diethylenetriamine and triphenylphosphine were used as fluorescence groups of Mito-Rh, respectively, to recognize and locate the groups. The fluorescence probe molecule Mito-Rh itself is colorless and fluorescence free, and when ATP is added, the ATP can interact with the probe molecule Mito-Rh, which makes the fluorescence group Rhodamine B lactam ring open, resulting in a strong red fluorescence of the probe molecule Mito-Rh at 583nm. The probe molecule Mito-Rh has a high sensitivity to ATP. The fluorescence increases 81 times before and after the reaction, and the detection range is 0.1-10 mm, which matches the content of ATP in mitochondria. At the same time, compared with other biological anions, such as UTP and UTP, the probe has a specific selectivity for ATP, which is mainly attributed to the double recognition site hydrogen bond interaction and 蟺-蟺 stacking of the probe to ATP. In addition, the probe can be accurately located in mitochondria and successfully used to monitor the changes of ATP content in mitochondria in real time. Naphthalene imide, nitrobenzodiazole and morpholine were used as fluorescence groups of Lyso-Nh, respectively, to recognize and locate the groups. The probe itself has the effect of solitary pair electrons of morpholine group N atom and the PET process produced by quenching group (recognition group). The probe has no fluorescence emission, and when H _ S _ 2S is added under pH=5 condition, The characteristic fluorescence emission peak of naphthalimide is produced at 536 nm by the recognition of the absence of the group and the protonation of the lone pair electrons of the N atom. The fluorescence enhancement was 72 times before and after the reaction, which indicated that the probe was sensitive. At the same time, the probe has good selectivity and is not affected by other biologically related substances. In addition, the probe can detect changes in H 2S content in lysosomes.
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3
本文编号：2185995
[Abstract]:Fluorescence analysis method has the advantages of high sensitivity, good selectivity, fast analysis speed, simple operation and easy in situ imaging of cells. It is widely used in the fields of biochemistry, environmental science and medical diagnosis. At the same time, Rhodamine B and naphthalimide fluorescent dyes are widely used because of their high fluorescence quantum yield, good photothermal stability, easy structure modification and low price. Therefore, in this thesis, Rhodamine B and naphthalimide were used to design and synthesize two fluorescent probes for the detection of ATP in mitochondria and H _ 2s in lysosome, respectively. The details are as follows: 1. The fluorescent probe molecules Mito-Rh and Lyso-Nhwere designed and synthesized, and their structures were confirmed by time-of-flight mass spectrometry (TF-MS), nuclear magnetic resonance (NMR) and nuclear magnetic resonance (NMR) carbon spectroscopy. Rhodamine B, diethylenetriamine and triphenylphosphine were used as fluorescence groups of Mito-Rh, respectively, to recognize and locate the groups. The fluorescence probe molecule Mito-Rh itself is colorless and fluorescence free, and when ATP is added, the ATP can interact with the probe molecule Mito-Rh, which makes the fluorescence group Rhodamine B lactam ring open, resulting in a strong red fluorescence of the probe molecule Mito-Rh at 583nm. The probe molecule Mito-Rh has a high sensitivity to ATP. The fluorescence increases 81 times before and after the reaction, and the detection range is 0.1-10 mm, which matches the content of ATP in mitochondria. At the same time, compared with other biological anions, such as UTP and UTP, the probe has a specific selectivity for ATP, which is mainly attributed to the double recognition site hydrogen bond interaction and 蟺-蟺 stacking of the probe to ATP. In addition, the probe can be accurately located in mitochondria and successfully used to monitor the changes of ATP content in mitochondria in real time. Naphthalene imide, nitrobenzodiazole and morpholine were used as fluorescence groups of Lyso-Nh, respectively, to recognize and locate the groups. The probe itself has the effect of solitary pair electrons of morpholine group N atom and the PET process produced by quenching group (recognition group). The probe has no fluorescence emission, and when H _ S _ 2S is added under pH=5 condition, The characteristic fluorescence emission peak of naphthalimide is produced at 536 nm by the recognition of the absence of the group and the protonation of the lone pair electrons of the N atom. The fluorescence enhancement was 72 times before and after the reaction, which indicated that the probe was sensitive. At the same time, the probe has good selectivity and is not affected by other biologically related substances. In addition, the probe can detect changes in H 2S content in lysosomes.
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3
【参考文献】
相关期刊论文 前2条
1 曹成波;朱艳丽;于学丽;张长桥;;萘酰亚胺类功能材料应用研究进展[J];精细与专用化学品;2007年Z1期
2 赵同丰,赵德丰,于华云,程侣柏;1,8-萘酰亚胺类荧光材料的进展[J];染料工业;1997年01期
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