Synthesis And Properties Of Reaction-Based Fluorescence Probes For Detection Of Biothiols

Biothiols play a very important role in promoting the metabolism of organisms.Their abnormal content is closely related to many diseases.At present,there are many methods that can be used to detect biological thiols.Among many methods,fluorescence analysis method is widely used due to less sample requirements,easy operation,and low cost.It is important to design and synthesize thiol fluorescent probes with good specificity and strong tissue penetration.Biothiols can undergo many nucleophilic reactions,and flavonoid derivatives and pyrazoline derivatives also have high biological activity and optical properties.Therefore,based on the nucleophilic reaction of biothiol,5 kinds of flavonoid derivatives fluorescent probes(HFL1-HFL5)for detecting Cys and 4 kinds of pyrazoline derivatives fluorescent probes for detecting GSH(HPY1-HPY4)were synthesized,the structures of probe compounds were characterized by HRMS,IR,1H NMR,13 C NMR,and its spectral properties were studied and calculated.At the same time,its mechanism was further verified by IR,NMR,MS and many other methods.Finally we explored the toxicity of the probe and cell imaging applications,and achieved staged results.The main content included the following:1.Using flavonoid derivatives as the precursor and acrylate as the targeting group,five fluorescent probes(HFL1-HFL5)were designed and synthesized for efficient detection of Cys.The spectroscopic research has confirmed that these five probes have good selectivity to Cys,and the optical phenomena have been reasonably explained by theoretical calculations.The identification mechanism is further confirmed by IR,NMR and MS.In pure PBS(p H7.4),the probe HFL1 still has a strong fluorescence response to Cys,while other probes of the same type have no fluorescence response in pure PBS(p H 7.4).In the PBS(10 m M,p H7.4,50% Et OH)system,the probes HFL4 and HFL5 have better fluorescence performance than the probes HFL2 and HFL3,and probe HFL5 shows a good specificity for Cys.Biological experiments have also shown that such probes are less toxic and can be used in living cells.2.Using pyrazoline as the precursor and acrylate as the targeting group,four fluorescentprobes(HPY1-HPY4)were designed and synthesized for efficient detection of GSH.Spectral performance experiments show that this type of probe has good selectivity to GSH.The probe itself has no fluorescence in a PBS(10 m M,p H 7.4,20% DMSO)system.When added GSH,the system appears strong blue fluorescence,so as to achieve the efficient detection of GSH.The identification mechanism is further confirmed by NMR,MS and theoretical calculations.Biological experiments have also shown that such probes are less toxic and can be used in living cells.