【作者】 张娇；

【导师】 赵红；

【作者基本信息】 东南大学 ， 化学工程与技术， 2020， 硕士

【摘要】 荧光探针法具有选择性好、灵敏度高和检测限低的分析优势,适于生物体内成像研究,广泛应用于重金属离子及生物活性物种的特异性检测。汞离子,铜离子和铝离子等金属离子,对社会生产活动有着重要影响,其含量的异常与生命过程中的一系列生理活动和疾病紧密相关。因此,对特异性识别重金属离子的荧光探针的探究成为研究重点。基于萘酰亚胺类化合物和罗丹明B类化合物具有良好的光学性能和易于修饰的化学结构:我们设计并合成了两种基于萘酰亚胺类衍生物的荧光淬灭型离子探针L和BNP1,探针L专一识别铜离子,探针BNP1特异性识别Cu²⁺（紫外和荧光双重响应）和Hg²⁺（荧光响应）;基于罗丹明类化合物独特的螺环开关转换结构,利用荧光共振能量转移机理（FRET）,设计了一种荧光增强型Cu²⁺探针Rh F。之后,为了改进具有罗丹明内酰胺结构的荧光探针的光学性质和提升探针体内成像的能力,我们在罗丹明母体上又引入了费舍尔醛来扩展罗丹明结构的共轭体系,使探针的吸收和发射波长大于700 nm,最终得到了一个多功能近红外类离子探针NB,可用来检测Cu²⁺,Hg²⁺和Al³⁺。相关化合物的结构均已通过质谱、核磁氢谱和核磁碳谱进行了表征。具体内容如下:1、设计和合成了两种基于萘酰亚胺类衍生物的高灵敏度和选择性的荧光探针L和BNP1,可用来检测实际水样和生物体活细胞中的Cu²⁺。探针L在CH₃CN/PBS（10 m M,9:1,v/v）对Cu²⁺具有很好的选择性。探针L加入Cu²⁺后,荧光几乎完全淬灭。L-Cu²⁺络合物2:1的化学计量比通过Job’s Plot图和ESI-MS进行了分析确认。¹H NMR滴定数据和密度泛函理论（DFT）研究也支持探针和Cu²⁺的作用机理。探针L具有良好的细胞通透性,并且已成功应用于A549细胞中Cu²⁺的成像研究。此外有关探针BNP1对铜离子/汞离子的识别行为也进行了详细研究,探针BNP1凭借高灵敏度、高选择性、检测限低、抗干扰能力强以及细胞毒性小的优势,成功实现实际水样和细胞中Cu²⁺的检测,制成的试纸条也可快速检测Cu²⁺。2、设计并合成了一种基于罗丹明衍生物的新型荧光探针Rh F。该探针是利用FRET机理设计而成,对Cu²⁺表现出比色、荧光比例响应和较大的斯托克斯位移（237 nm）,并且不受其他共存金属离子的干扰。该探针对Cu²⁺的潜在应用也可以通过试纸条来完成。此外,探针Rh F还可用于检测实际水样中的Cu²⁺。3、研究了基于罗丹明内酰胺平台设计的近红外类离子型荧光探针NB在DMF和水的混合溶液中特异性识别Cu²⁺（紫外响应）、Hg²⁺（紫外和荧光双重响应）和Al³⁺（荧光响应）的响应行为、识别机理以及细胞成像研究。探针NB抗离子干扰能力强,p H值适用范围大且具有较低的检测限,低至纳摩尔级别。同时探针具有较低的细胞毒性,已经成功应用于A549细胞中的Hg²⁺和Al³⁺成像研究。更多还原

【Abstract】 Fluorescent probes are widely used for the specific detection of some heavy metal ions and biologically active species due to their unique advantages such as high selectivity,high sensitivity,and suitability for in vivo imaging studies.Metal ions have an important impact on social production activities,such as mercury ions,copper ions and aluminum ions.The abnormal content of these ions is closely related to a series of physiological activities and diseases in life.Therefore,the research on fluorescent probes that specifically recognize heavy metal ions has become a hot spot.A lot of fluorescent probes are designed and synthesized based on the excellent optical properties and easily modified chemical structures of 1,8-naphthimide and rhodamine B fluorophiles.There are two types of fluorescence quenching probes L and BNP1 based on naphthalimide derivatives,probe Rh F is designed based on fluorescence resonance energy transfer and the unique structure of rhodamine B derivatives,and a multi-functional near-infrared probe NB is designed based on the expanded rhodamine B fluorophore.The structures of related compounds have been characterized by MS,¹H NMR and ¹³C NMR.The specific contents are as follows:1.Two highly sensitive and selective fluorescent probes L and BNP1 based on 1,8-naphthimide derivatives are designed and synthesized to detect copper ion in domestic water samples and living cells.Probe L has good selectivity for Cu²⁺in CH₃CN/PBS（10 m M,9:1,v/v）.After the addition of Cu²⁺,the fluorescence of probe L is almost completely quenched.The2:1stoichiometric ratio of L-Cu²⁺complex is confirmed by Job’s diagram and ESI-MS.Further evidence of the interaction between the probe and Cu²⁺is provided by ¹H NMR titration data and density functional theory（DFT）studies.Probe L with good permeability has been successfully applied to the imaging of Cu²⁺in A549 cells.Similarly,the identification behavior of probe BNP1and copper/mercury ion is also studied in detail.Probe BNP1 has been successfully applied to the detection of Cu²⁺in water and A549 cells with high sensitivity,high selectivity,low detection limit,strong anti-interference ability and low cytotoxicity.And it has also been successfully made into strips that can quickly detect copper ions.2.A novel fluorescent probe Rh F based on rhodamine B derivative is designed and synthesized.Based on FRET mechanism,the probe shows a ratiometric fluorescence response to Cu²⁺and a large stokes displacement（237 nm）.And the recognition process is not disturbed by other co-existing metal ions.The potential application of the probe to Cu²⁺can be accomplished by strip paper.In addition,the probe can be used to detect Cu²⁺in water samples.3.The response behavior,recognition mechanism and cell imaging of near-infrared fluorescent probe NB based on rhodamine lactam platform in DMF and H₂O mixture are studied.The probe specifically identify Cu²⁺（UV response）,Hg²⁺（UV and fluorescence response）and Al³⁺（fluorescence response）.The probe NB has strong anti-ion interference ability,wide application range of p H value and low detection limit,as low as nanomole level.At the same time,the probe with low cytotoxicity has been successfully used in the imaging of Hg²⁺and Al³⁺ions in A549 cells.更多还原