【作者】 王林；

【导师】 籍雪平；

【作者基本信息】 河北医科大学 ， 药物分析， 2023， 博士

【摘要】 电化学发光（ECL）技术已经成为现代药物分析领域的有效技术。因其具有背景信号低,实验操作简单及其高特异性等优点,特别适用于难以检测、组分复杂的体系中。近年来已经被广泛用于临床分析、癌症诊断、食品检测以及药物分析等领域。ECL传感器是结合了化学发光和电化学的优点,能够将目标分子和电极上的识别元件通过化学作用转换成不同强度的ECL发光信号,并根据信号的强弱变化实现对目标物的定性和定量检测的技术手段。但是,传统的ECL传感器已经无法满足微量甚至痕量待测物的灵敏且特异性检测。因此,寻求提高发光体的ECL效率和识别元件对目标物的高特异性成为构建灵敏ECL传感器的重要部分。本文的研究内容主要为:第一部分:基于钌衍生物和硫化镉量子点之间的共振能量转移构建电化学发光传感器用于绿原酸的检测本实验设计了一种基于共振能量转移（RET）策略的高灵敏电化学发光传感器用以检测绿原酸。传感器引入了经典发光体二维钌衍生物纳米片（Ru@Zn-MOF）作为能量的受体,L-半胱氨酸包裹的硫化镉量子点（L-Cd S QDs）作为能量的供体,两者的相互作用显示了优异的ECL响应。我们对此体系的反应机理进行了详细讨论。将L-Cd S QDs通过逐层修饰的方法直接滴加在MOF纳米片表面,不仅可以减小供体和受体之间能量转移的距离,同时降低了传输过程中的能量损失,提高ECL发光信号。同时,将导电性良好的碳纳米管和石墨烯复合物（CNT@GO）引入传感器可以加速对共反应剂S₂O₈^2-的催化,实现了对L-Cd S QDs/Ru@Zn-MOF/S₂O₈^2-体系ECL信号的进一步放大。基于最优条件下,所构建的传感器对目标物绿原酸的检测范围是1.0×10^-10～1.0×10^-4 mol·L^-1,检测限是3.2×10^-11 mol·L^-1。此外,本传感器对CGA表现了优异的稳定性和选择性。第二部分:基于g-C₃N₄@Cu O构建“signal off-on”型电化学发光传感器用于芦丁的检测本研究以类石墨相氮化碳纳米片（g-C₃N₄）为发光体（能量供体）,Cu O纳米颗粒为猝灭探针（能量受体）,三元合金Au Pt Ag纳米粒子为共反应促进剂,MIL-101型MOF作为发光体的载体,构建了基于ECL-RET的“signal off-on”型电化学发光传感器用于对黄酮类药物芦丁（Rutin）的灵敏检测。本实验中,我们通过原位合成的方式将猝灭探针Cu O掺杂到g-C₃N₄表面,提高了RET的转化效率。当没有待测物时,Cu O通过RET的方式抑制了发光体g-C₃N₄的ECL强度（signal off）,加入待测物Rutin后,Cu O会被还原从而在发光体表面脱落,ECL强度得以恢复（signal on）。基于最优条件下,所构建的传感器对目标待测物芦丁的检测范围是10 n M～50μM,检测限是1.7 n M。同时,该传感器显示出具有良好的稳定性和重现性。第三部分:基于自增强钌衍生物和类石墨氮化碳的比率型电发光传感器检测谷胱甘肽传统的电致发光体系是不同类型的发光体和共反应剂之间的相互作用,实现信号放大。但往往两者之间的分子间作用存在电子传输距离远,能量损失过高的问题。本实验是将阳极共反应剂三乙胺（TEA）与发光体钌配合物Ru（dcbpy）₃²⁺制备到MOF-5中,MOF-5既可以作为载体均匀分散发光物质,同时作为介质将TEA和Ru（dcbpy）₃²⁺交联到一起减小了电子传输距离,放大阳极ECL信号响应。另一方面,采用g-C₃N₄作为另一信号探针,以S₂O₈^2-为阴极共反应剂,构建了双向捕获目标物的ECL比率传感器实现对谷胱甘肽（GSH）的高灵敏检测。在最优条件下,所设计的传感器对GSH的检测范围是25 p M～50 n M,检测限为8.5 p M。以Ru（dcbpy）₃²⁺/TEA和g-C₃N₄/S₂O₈^2-分别为阳极和阴极信号探针并结合RET策略,为GSH的特异性检测提供了一个行之有效的方法。第四部分:基于鲁米诺衍生物和金纳米簇的复合物设计比率型电化学发光传感器用于肾上腺素的检测本实验利用谷胱甘肽包裹的金纳米簇（GSH-Au NCs）作为检测信号,N-（4-氨丁基）-N-乙基异鲁米诺（ABEI）分子作为参比信号,将GSH-Au NCs和ABEI通过酰胺键交联在一起,不仅对ABEI的易溶解性起到抑制作用,同时提高了阳极信号的稳定性。常温下合成了Ce MOFs,将其作为双发光体复合材料的载体改善传感界面的分散性,实现了对目标物肾上腺素（EP）的灵敏检测。作为儿茶酚类物质,EP的出现可以猝灭GSH-Au NCs的阴极信号,而ABEI作为参比信号,可以进一步避免检测体系中其他组分的干扰。利用阴极ECL信号与阳极参比信号的比值,本实验设计了双信号输出的比率型传感器(E_cathode/E_anode),实现对EP的灵敏检测。在最优条件下,该传感器对EP的检测范围是30 n M～10μM,检测限为14 n M。此方法为检测EP含量提供了一个选择性和灵敏度更高的平台,在药物分析中显示出优异的实用价值。更多还原

【Abstract】 Electrochemiluminescence（ECL）has become an effective technique in the field of modern pharmaceutical analysis.As a powerful analytical technique,ECL method exhibited lots of advantages such as the low background,simple operation and extremely high selectivity.The ECL sensor especially applicable for the systems that are difficult to detect or it contains complex components.In recent years,it has been widely used in clinical analysis,cancer diagnosis,food detection and pharmaceutical analysis.ECL sensor combining the advantages of chemiluminescence and electrochemistry,which can convert the chemical reaction between model analyte and the identification module into series of ECL intensity signals,and realize the qualitative and quantitative detection of the model analyte according to the changing ECL intensity.However,the traditional ECL sensors have been unable to achieve the sensitive and specific detection of model analyte.Therefore,seeking to improve the ECL efficiency and the specificity between recognition module and target molecule have become important parts in the construction of highly sensitive ECL sensors.The main research of this paper is divided into the following parts:Part One:Electrochemiluminescence resonance energy transfer system between ruthenium-based nanosheet and Cd S quantum dots for detection of chlorogenic acidThe study proposed a new strategy for ultrasensitive detection of chlorogenic acid（CGA）by fabricating a resonance energy transfer（RET）sensing platform.Herein,we designed the novel system by introducing ruthenium-based 2D metal-organic framework nanosheets（Ru@Zn-MOF）as energy acceptor and L-cysteine capped Cd S quantum dots（L-Cd S QDs）as energy donor,which exhibited good ECL response.The possible mechanism of the modified electrode surface reaction was discussed.Modifying appropriate of L-Cd S QDs directly on ultrathin MOF nanosheets greatly shorten the electron-transfer distance and reduce energy loss,therefore significantly improving the ECL efficiency.According to the optimal conditions,the constructed sensor for the detection of CGA exhibited a wide linear range from 1.0×10^-10 to 1.0×10^-4 mol·L^-1 with a low detection limit of3.2×10^-11 mol·L^-1.Besides,the prepared sensor demonstrated good stability and highly selective detection of target molecule.Part Two:A novel“signal off-on”electrochemiluminescence sensor based on g-C₃N₄@Cu O for rutin detectionIn this study,a high efficiency“signal off-on”ECL sensor was constructed for ultrasensitive detection of rutin based on carboxylated graphitic carbon nitride（g-C₃N₄）as luminophores（energy donor）,Cu O nanoparticles as quenching probe（energy acceptor）,tri-metallic Au Pt Ag as coreaction accelerator and MIL-101 type MOF as carrier of luminophores.In present platform,Cu O NPs in situ grew on the surface of g-C₃N₄,greatly shorten the distance between energy acceptor and donor pair and greatly enhancing the RET conversion efficiency.In the absence of rutin,Cu O NPs inhibits the ECL intensity of g-C₃N₄（signal off）through RET strategy.Upon the addition of rutin,Cu O NPs could be reduced by rutin and fall off on the surface of g-C₃N₄,the ECL intensity was significantly recovered（signal on）.Under the optimal conditions,the constructed ECL-RET sensor realized the sensitive detection of rutin range from 10 n M to 50μM with the detection limit of 1.7 n M.Furthermore,the as-prepared ECL-RET system exhibits good stability and reproducibility.Part Three:A ratiometric electrochemiluminescence sensor for glutathione detection based on ruthenium derivatives and g-C₃N₄The traditional electrochemiluminescence system is the interaction between different types of luminophores and coreactant to realize signal amplification.However,the intermolecular interaction between them has the drawbacks such as long electron transport distance and high energy loss.In this study,the anodic coreactant trimethylamine（TEA）and the Ru-based luminophores(Ru（dcbpy）₃²⁺)were prepared in the surface of MOF-5.MOF-5could not only be used as carrier to disperse luminophores evenly,but also as the medium to cross-link TEA and Ru（dcbpy）₃²⁺through amide reaction,reducing the electron transport distance and amplifying the anode ECL signal response.On the other hand,using g-C₃N₄ as cathode signal probe and S₂O₈^2-as coreactant,a ratiometric ECL sensor was constructed to achieve highly sensitive detection of glutathione（GSH）.Under the optimal conditions,the detection range of the designed sensor for GSH is 25 p M～50 n M with the detection limit is 8.5 p M.Introducing Ru（dcbpy）₃²⁺/TEA and g-C₃N₄/S₂O₈^2-as anode and cathode signal probe separately,and combining with RET strategy,the sensor provides an effective method for highly sensitive and specific detection of GSH.Part Four:A ratiometric electrochemiluminescence sensor based on luminol derivatives and gold nanoclusters for the epinephrine detectionHerein,a sensitive electrochemiluminescent platform based on N-（4-aminobutyl）-N-ethylisoluminol（ABEI）and glutathione coated gold nanoclusters（GSH-Au NCs）as double luminophores is prepared.Here,GSH-Au NCs were used as working signals,and ABEI were used as reference signals.The crosslinking of GSH-Au NCs and ABEI through amide reaction not only inhibited the solubility of ABEI,but also improved the stability of anode signal response.Ce MOFs were synthesized at room temperature and used as the carrier of double luminescent composites which could improve the dispersion of the sensor interface.The dual signal electrochemiluminescent sensor was used to detect epinephrine（EP）sensitively.As catecholic substances containing phenolic hydroxyl,the presence of EP can quench the cathode signal from GSH-Au NCs.While ABEI,as a reference signal,can further avoid the interference of other components in the detection system.Based on the ratio between the cathode ECL signal and the anode reference signal,a dual signal output ratiometric sensor(E_cathode/E_anode)is designed in this study.Under optimal conditions,the detection range of EP is 30 n M～10μM,and the detection limit is 14 n M.This method provides a more selective and sensitive platform for EP detection and shows excellent practical value in pharmaceutical analysis.更多还原