| Heavy metal is a kind of metal with larger density,which is a common pollutant,topically introduced by industrial pollution,the traffic exhaust as well as food and cosmetics.Heavy metal can be concentrated along with the food chain or accumulated in living oganism due to its feature of high lipid solubility and easy chelating ability,so that causes regression function damage of organism and biosystem once exceeds a normal level.In addition to the traditional detection methods,the emerging detection technology includes small molecule fluorescent probes.However,these methods can't be applied in vivo detection because their intrinsic limitation.In the current work,we aim at developing a bioluminescent method for the detection of heavy metals including paladium and mercury.Bioluminescence is a specific chemiluminescence phenomenon in living organisms,depending on biological processes that widely distribute in nature.The nature of bioluminescence is that the interaction of enzyme and substrate will release light after a series of biological responses.With the in-depth research,bioluminescent imaging technology has been widely applied in various fields and becomes a very important detection considering its high sensitivity,reasonable biocompatibility and ideal visualization characteristics.Among various bioluminescent systems,the firefly bioluminescent system is the most studied so far.Based on the existing mechanism of palladium ions and mercury ion fluorescent probe,two classes of five bioluminescent probes for palladium and mercury were well designed and synthesized for in vivo and in vitro detection,respectively.This topic contains three parts:the preface,palladium ion detection method and mercury ion detection method.The first part of this thesis provides a general introduction on heavy metal ions and their pollution and toxicity research status,as well as detection methods,including:current small molecule fluorescence probes of palladium and mercury ions,the basic detection principle,and the general background of bioluminescent and firefly luciferase system.The second part mainly introduces two bioluminescent probes for palladium detection.Widely used in the jewelry,auto electronic,medical,fuel cell and catalysis industry,palladium ion may cause a great harm once enters into body.Using propargyl ether as the hydrolysis group,two palladium probes can be catalyzed by palladium to release the substrate of luciferase-luciferin,so as to realize the purpose of detection of palladium.This part discusses the synthesis and in vitro and in vivo application of probes.The biological evaluation results confirms the feasibility of linear detection in vitro,and the real-time monitoring of palladium ions in mice.The third part mainly presents three bioluminescent probes for mercury recognition.Mercury is a very important heavy metal that applied in various industries,including manufacturing industry,pharmaceutical industry,cosmetic industry and mining industry,etc.Due to its toxicity,there are a lot of clinical poisoning cases of mercury.With great development of industry,massive pollution of mercury ion have often occurred.This part privides three mercury probes CHG-1,CHG-2 and CHG-3.CHG-1 was using acetylene butyl ether and CHG-2/3 were using vinyl ether as hydrolysis group.In brief,they can release the luciferin under the reaction with mercury for the detection of mercury ion.This part discusses the synthesis of three probes,and its detection application in vitro and in vivo..The experimental results demonstrates that CHG-3 can only be limited to in vitro detection because of its reaction temperature,and CHG-2 has a narrow determination range with poor sensitivity,so we chose CHG-2,which is more sensitive with broad testing range,to discuss in vivo.These promising results indicate CHG-2 can be implemented to detect the mercury poisoning in vivo,and the potential value to detect the accumulation state of mercury poisoning... |
PDF Full Text Request