水溶性不对称吲哚菁染料的合成、光学性能及生物标示

发布时间：2018-04-10 20:11

本文选题：吲哚菁染料 + 非离子水溶性树权PEG醚链　；参考：《河南大学》2014年硕士论文

【摘要】：目的：吲哚菁染料具有光谱范围广(550nm-1200nm)、荧光量子产率高和检测灵敏度高等特点,现已被广泛应用于荧光探针、蛋白质标示、DNA测序和细胞染色等方面。我们设计在吲哚“N”位引入非离子型水溶性树杈PEG醚链,得到拥有产率更高、水溶性更好、光学性质更加优良等特点的新型水溶性不对称吲哚菁染料。方法：吲哚菁染料一般通过三个步骤合成得到,首先：通过Fisher反应得到碱性杂环；其次：碱性杂环与烷基化试剂反应得到季铵盐；最后：缩合剂与带有活性甲基的季铵盐反应得到目标产物。磺酸基苯肼、对甲苯磺酰氯和聚三乙二醇等为原料合成得到一种新型水溶性不对称吲哚五甲川菁染料Cy5-671和一种新型水溶性不对称吲哚三甲川菁染料Cy3-567。由磺酸基苯肼和对溴甲基苯甲酸等为原料合成得到一种吲哚五甲川菁染料Cy5-668。利用紫外可见分光光度计和荧光分光光度计测定了产物的光学性质；吲哚菁染料经过琥珀酰亚胺酯活化后与牛血清白蛋白或羊抗小鼠IGg进行了标示实验；同时把Cy5-671和Cy5-668的性质进行了对比。结果：我们总共合成十余种中间产物,最终得到了三种水溶性不对称吲哚菁染料,其结构都经'HNMR和MS确认。Cy5-671的最大吸收波长为657nm,消光系数为1.6×105／M-1cm-1,最大荧光发射波长671nm,荧光量子产率为0.24；Cy5-668的最大吸收波长为650nm,消光系数为1.4×105/M-1cm-1,最大荧光发射波长668nm,荧光量子产率为0.10；Cy3-567的最大吸收波长为559nm,消光系数为0.8×105/M-lcm-1,最大荧光发射波长567nm,荧光量子产率为0.06；Cy5-671在水中的光稳定明显高于Cy5-668；Cy5-671标记牛血清白蛋白的D/P值1.96,Cy5-668标记牛血清白蛋白的D/P值1.57,Cy3-567标记牛血清白蛋白的D/P值1.59。结论：新型水溶性不对称吲哚五甲川吲哚菁染料Cy5-671在吲哚“N”位引入非离子型水溶性树权PEG醚链,与Cy5-668相比较,Cy5-671拥有较好的水溶性,并且容易分离提纯,产率也较高；拥有较好的光谱性质和较高的荧光量子产率并且标示蛋白质能获得比较好的有利于实际应用的D/P值。
[Abstract]:Objective: indocyanine dyes have been widely used in fluorescent probes, protein-labeled DNA sequencing and cell staining for their high fluorescence quantum yield and high detection sensitivity, with a wide spectrum range of 550 nm ~ 1 200 nm.We designed a new water-soluble asymmetric indocyanine dye with higher yield, better water solubility and better optical properties by introducing non-ionic water soluble branch PEG ether chain into indole "N" site.Methods: indocyanine dyes were synthesized in three steps: first, basic heterocycles were obtained by Fisher reaction, secondly, quaternary ammonium salts were obtained by the reaction of basic heterocycles with alkylation reagents.Finally, the reagents reacted with quaternary ammonium salts with active methyl to obtain the target product.A new water-soluble asymmetric indolepentacyanine dye Cy5-671 and a new water-soluble asymmetric indoletrimethylcyanine dye Cy3-567 were synthesized from sulfonated phenylhydrazine, p-toluenesulfonyl chloride and polyethylene glycol.An indole-pentamethylcyanine dye Cy5-668 was synthesized from sulfonated phenylhydrazine and p-bromomethyl benzoic acid.The optical properties of the product were determined by UV-Vis spectrophotometer and fluorescence spectrophotometer, and the indocyanine dyes were labeled with bovine serum albumin (BSA) or sheep anti-mouse IGg after the activation of succinimide.At the same time, the properties of Cy5-671 and Cy5-668 are compared.Results: we synthesized more than ten intermediate products and finally got three water-soluble asymmetric indocyanine dyes.The D / P value of bovine serum albumin labeled with D- P value of 1.57 Cy3-567 was 1.59.Conclusion: the new water-soluble asymmetric indole-pentacanine indoline cyanine dye (Cy5-671) was introduced into the indole-" N "site with a non-ionic water-soluble tree weight PEG ether chain. Compared with Cy5-668, Cy5-671 has better water solubility, easy separation and purification, and higher yield.It has better spectral properties, higher fluorescence quantum yield and better D / P value for practical application.
【学位授予单位】：河南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R914.5

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