基于能量转移的化学发光信号放大分析体系构建及检测研究

发布时间：2019-03-08 20:00

【摘要】：化学发光法由于其检测过程具有无背景光干扰且方法灵敏度高的优点,常被用于分析化学领域。其中双(2,4,6-三氯苯基)草酸酯-过氧化氢体系由于量子产率高、杂质干扰小以及可以在接近中性pH条件下进行化学发光反应,不仅可以对小分子进行分析检测,而且也适合对生物分子进行分析检测。介孔二氧化硅作为一种纳米材料,具有较大的孔容、可调的孔道结构、表面基团易于被修饰以及良好的生物相容性等特点,在生物和分析领域应用广泛。氧化石墨烯量子点(GOQDs)不仅保持了GO原有的生物相容性、低毒性等优异特点,还展现出很强的化学发光增敏作用。本文主要基于化学发光共振能量转移体系,完成了一些化学发光分析检测方法的构建以及应用。具体概述如下:(1)以化学发光共振能量转移体系为基础,利用以单链DNA作为门控且用GOQD包裹的介孔二氧化硅合成了一种具有三重信号放大机制的生物传感器,分别采用傅里叶变换红外光谱仪、紫外-可见分光光度法、Zeta电位等方法进行了表征。实验结果表明,当miRNA-21的浓度在0.005~50 pmol L~(-1)范围内,该体系的化学发光相对强度与mi RNA-21的浓度的对数存在很好的线性关系,检出限为0.002 pmol L~(-1),实际样品(人体血清样品)加标回收率为93.6%-108.8%。(2)利用氧化石墨烯量子点(GOQD)的化学发光增敏特性,构建一种4-硝基苯酚的超灵敏、高选择性的检测方法。实验结果表明,当4-硝基苯酚的浓度在1.0~300 pmol L~(-1)范围内,该体系的化学发光相对强度与4-硝基苯酚的浓度存在很好的线性关系,检出限为0.3 pmol L~(-1),实际样品(自来水和湘江水)加标回收率为93.4%-107.9%。(3)以化学发光共振能量转移体系为基础,结合氧化石墨烯(GO)的化学发光猝灭作用,利用谷胱甘肽(GSH)的还原氧化石墨烯的特性,使化学发光恢复,构建了一种简单、快捷、灵敏的检测谷胱甘肽的方法。实验结果表明,当谷胱甘肽的浓度在0.1~200 nmol L~(-1)范围内,该体系的化学发光相对强度与谷胱甘肽的浓度存在很好的线性关系,检出限为30 pmol L~(-1),实际样品(人体血清样品)加标回收率为93.0%-103.0%。
[Abstract]:Chemiluminescence method is often used in analytical chemistry because of its advantages of no background interference and high sensitivity. The bis (2,4,6-trichlorophenyl) oxalate-hydrogen peroxide system, due to its high quantum yield, low impurity interference and chemiluminescence reaction under near neutral pH conditions, can not only analyze and detect small molecules, It is also suitable for analysis and detection of biomolecules. Mesoporous silica, as a kind of nano-material, has the characteristics of large pore volume, adjustable pore structure, easy modification of surface groups and good biocompatibility. It has been widely used in biological and analytical fields. Graphene oxide quantum dots (GOQDs) not only maintain the original biocompatibility and low toxicity of GO, but also exhibit strong chemiluminescence sensitization. In this paper, based on the chemiluminescence resonance energy transfer system, some chemiluminescence analysis and detection methods have been constructed and applied. The main contents are as follows: (1) based on the chemiluminescence resonance energy transfer system, a biosensor with triple signal amplification mechanism was synthesized by using mesoporous silica coated with GOQD and gated with single strand DNA. Fourier transform infrared spectroscopy, UV-vis spectrophotometry and Zeta potential were used respectively. The experimental results show that there is a good linear relationship between the relative intensity of chemiluminescence and the logarithm of miRNA-21 concentration when the concentration of miRNA-21 is in the range of 0.005 ~ 50 pmol L ~ (- 1), and the detection limit is 0.002 pmol / L ~ (- 1). The standard recovery of the actual sample (human serum sample) was 93.6% ~ 108.8%. (2) the chemiluminescent properties of graphene oxide quantum dot (GOQD) were used to construct a 4-nitrophenol hypersensitivity. A highly selective detection method. The results show that when the concentration of 4-nitrophenol is in the range of 1.0 ~ 300 pmol L ~ (- 1), there is a good linear relationship between the relative intensity of chemiluminescence and the concentration of 4-nitrophenol, and the detection limit is 0.3 pmol / L ~ (- 1). The standard recovery of actual samples (tap water and Xiangjiang water) was 93.4% ~ 107.9%. (3) on the basis of chemiluminescence resonance energy transfer system, combined with the chemiluminescence quenching effect of graphene oxide (GO), Based on the reduction of graphene oxide by glutathione (GSH), a simple, rapid and sensitive method for the determination of glutathione was developed to restore chemiluminescence. The results showed that there was a good linear relationship between the relative intensity of chemiluminescence and the concentration of glutathione in the range of 0.1 脳 200 nmol / L ~ (- 1) of glutathione, and the detection limit was 30 pmol / L ~ (- 1). The standard recovery rate of actual sample (human serum sample) was 93.0% ~ 103.0%.
【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

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