| Fluorescence probe is a molecular measuring device based on spectro-chemistry and optical waveguide and measurement technology,which can transform chemical information of analytical object into easily measured fluorescence signal.It is often used for labeling and quantitative analysis of nucleic acids and proteins.In this paper,the design,synthesis and application of two fluorescent probes for human serum albumin and mercury ion are discussed.Human serum albumin(HSA)is a biological macromolecule with important physiological functions.Being the most abundant transport protein in human blood plasmaa,it can adjust osmotic pressure,transport nutrients,metabolites and different series of exogenous drugs,so it plays a vital role in the pharmacokinetics and efficiency of drugs.Abnormal HSA levels are associated with coronary heart disease,multiple myeloma,diabetes,nephropathy,neurometabolic disorders,liver cirrhosis and other diseases.In addition,it is clinically used for symptomatic treatment of massive hemorrhage,shock,burn,erythroleukocytosis and hypoalbuminemia.Therefore,accurate and quantitative detection of HSA has extremely important research and application value in biological science,molecular biology,clinical medicine and other fields.In the second chapter of this thesis,we designed and synthesized a fluorescent probe RhHSA based on rhodanine and bromopropyne for the detection of HSA.RhHSA enhances its self-fluorescence by specific binding with drug binding site IIA,IIIA of HSA,and the fluorescence intensity showed a good linear relationship with the HSA concentration within the range of 0-0.5mg/mL(R2=0.9878),and the detection limit was 5.0 nM,about 3.33 mg/L.Compared with other biological macromolecules or common amino acids,RhHSA exhibits excellent selectivity to HSA,even can discriminates HSA from bovine serum albumin(BSA).In addition,RhHSA was successfully applied to the detection of HSA in rat urine samples and MCF-7 cells.The binding mechanism of fluorescence probe and HSA is clarified clearly by HSA's guanidine hydrochloride,trypsin denaturation test,competitive inhibition test and molecular docking simulation.In recent years,heavy and transition metal(HTM)ions have attracted much attention due to their harmful effects on the environment and biological systems.Among these HTM ions,mercury is a highly toxic substance that can cause harm to humans and the environment even at very low concentrations.Hg2+ can enter biological membranes and accumulate in the human body through the food chain,causing serious damage to health,including minamata disease,deafness and cognitive impairment.Therefore,developing an economical,simple,highly selective and sensitive determination method for the detection of Hg2+ content in the environment,food and biology has become an urgent research topics.In the third chapter,we designed and synthesized a kind of fluorescence probe,DAC-Hg,with coumarin as the parent nucleus for detecting Hg2+.Mercaptan protected aldehyde was used as the recognition group.The response mechanism of Hg2+ and DAC-Hg was verified by hydrogen spectrum,carbon spectrum,high resolution mass spectrometry,infrared spectrum and ultraviolet absorption spectrum.Compared with other common metal ions,DAC-Hg shows good selectivity and sensitivity to Hg2+.After the reaction between the probe DAC-Hg and Hg2+,the fluorescence was quenched,and the fluorescence intensity showed a good linear relationship with the Hg2+ concentration within the range of 0-10 eq(R2 =0.9962),with the detection limit of 5.0 nM,about 0.001 mg/L.Naked-eye evaluation with test strips demonstrated the potential of DAC-Hg for conveniently handled in-situ detection.The application of this established method on analyzing water environmental and seafood samples supplied satisfactory results,which provided an idea for the detection of other heavy metal ions and transition metal ions. |
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