检测二氧化硫衍生物、次氯酸根的荧光探针的设计合成及生物成像应用

发布时间：2018-04-24 02:09

本文选题：荧光探针 + 半菁染料　；参考：《云南师范大学》2017年硕士论文

【摘要】：二氧化硫衍生物常被做为食物防腐剂而使用在日常生活当中,用来防止食物氧化和细菌生长。随着工业化的进展,促使煤、石油以及化石燃料的大量使用,导致了SO2被大量排放到空气中。低浓度的二氧化硫衍生物对人体影响不大,然而,当二氧化硫衍生物的浓度超过一定范围,便会对人体健康产生一定损害,如:呼吸系统疾病、癌症等。次氯酸是人体在自然氧化代谢过程中产生的一种副产物,次氯酸与生物体的各项生理和病理过程有着密切的关系。当生物体内次氯酸浓度过高就会引起癌症、关节炎等疾病。因此,利用荧光探针技术定量检测二氧化硫衍生物和次氯酸根具有重要的意义。本论文设计和合成了用于检测亚硫酸根(T1和T2)、亚硫酸氢根(T3)和次氯酸根(T4)的荧光探针,并对它们的光物理性质和生物学应用进行了分析研究。研究发现,这几种探针都具有优异的选择性能,以及良好的光谱信号响应变化,这些探针被成功的应用于检测活体细胞和秀丽线虫中的目标分子。本论文主要包括以下内容:1、探针T1和探针T2是以香豆素做为荧光团,用来检测亚硫酸根的荧光探针。探针T1和T2分别在CH3CN-HEPES(0.02 M,pH 7.4)(V/V=2:8)的缓冲溶液,CH3CN-PBS(0.02 M,pH 7.4)(V/V=2:8)的缓冲溶液中进行测试。探针T1对亚硫酸根表现出较好的选择性,探针T1还被成功的应用到Hela细胞中对亚硫酸根检测。2、探针T3通过半菁染料和N-乙基-3,6-二甲醛基咔唑缩合得到。在光物理性质的研究中,我们观察到探针T3在DMF-PBS(0.01 M,pH=7.4)(V:V=3:7)缓冲液中加入HSO3-后,溶液的颜色发生了从淡黄色变成了无色的变化,并伴随着荧光增强的响应。探针T3被成功的应用到Hela细胞和秀丽线虫当中,通过荧光强度的改变定量的测定外源性HSO3-的含量。3、探针T4以探针T3做为基础,通过T3中的醛与盐酸羟氨作用生成肟,用来检测次氯酸根。T4在DMF-PBS(0.01 M,pH 7.4)(V/V=4:6)的缓冲溶液中测试,加入次氯酸根后,出现荧光增强的响应,在各种干扰离子的存在下,探针T4表现出优异的选择性。探针T4被成功地应用到生物荧光成像检测Hela细胞和秀丽线虫中的次氯酸跟离子,通过荧光强度的变化可以测定外源性ClO-的含量。
[Abstract]:Sulfur dioxide derivatives are often used as food preservatives in daily life to prevent food oxidation and bacterial growth. As industrialization progressed, the use of coal, oil and fossil fuels increased, causing SO2 to be pumped into the air. The low concentration of sulfur dioxide derivatives has little effect on human body. However, when the concentration of sulfur dioxide derivatives exceeds a certain range, it will cause certain harm to human health, such as respiratory diseases, cancer and so on. Hypochloric acid is a by-product produced by human body during natural oxidation and metabolism. Hypochlorite is closely related to various physiological and pathological processes of organism. When the concentration of hypochloric acid in organisms is too high, it can cause cancer, arthritis and other diseases. Therefore, quantitative detection of sulfur dioxide derivatives and hypochlorite by fluorescence probe technique is of great significance. In this paper, fluorescent probes were designed and synthesized for the detection of T1 and T2H _ 2, T3) and T4), and their photophysical properties and biological applications were studied. It is found that these probes have excellent selectivity and good spectral signal response. These probes have been successfully used to detect target molecules in living cells and nematodes. The main contents of this thesis are as follows: probe T1 and probe T2 are fluorescent probes using coumarin as fluorescence group to detect sulfite radical. The probe T1 and T2 were tested in the buffer solution CH3CN-PBSN 0.02 MN pH 7.4 / V / V 2: 8, respectively, in the buffer solution Ch _ 3CN-PBS _ 2: 8 (pH 7.4 / V / V _ (2: 8)). Probe T1 showed good selectivity for sulfite, and probe T1 was also successfully applied to detect sulfite in Hela cells. Probe T3 was obtained by condensation of hemicyanine dye with N-ethyl-3-dimethylcarbazole. In the study of photophysical properties, we observed that the color of the solution changed from light yellow to colorless after the addition of HSO _ 3- in the buffer solution of DMF-PBS(0.01 M _ 3 pH = 7.4 / V: v _ (3: 7), which was accompanied by the response of fluorescence enhancement. Probe T3 was successfully applied to Hela cells and nematodes. The content of exogenous HSO3- was determined quantitatively by changing the fluorescence intensity. The probe T4 was based on probe T3 and formed oxime through the reaction of aldehyde in T3 with hydroxylamine hydrochloride. The method was used to detect hypochlorite in DMF-PBS(0.01 pH 7.4 V / V 4: 6 buffer solution. After adding hypochlorite, the fluorescence response was enhanced. In the presence of various interference ions, the probe T4 showed excellent selectivity. The probe T4 was successfully applied to the detection of hypochloric acid and ions in Hela cells and nematode, and the content of exogenous ClO- could be determined by the change of fluorescence intensity.
【学位授予单位】：云南师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3

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