Fluorescent Probe Of H₂O₂ Based On The New Detection Site:Synthesis And The Application

Hydrogen peroxide plays adistinct role in biological systems and acts as an important messenger for cellular growth,proliferation and differentiation.Elevate hydrogen peroxide has been correlated with aging,Alzheimer‘s disease and cancer.Thus,it is of great importance to determine the hydrogen peroxide levels in living biological systems.Upto now,many measurement technologies have been developed to detect hydrogen peroxide,such as electrochemical method,ESR method and HPLC method.However,traditional methods can not detect the concentration of hydrogen peroxide in living cells.A key advantage of fluorescent probes over traditional methods is the ability to detect hydrogen peroxide in living cells.Therefore,we designed two fluorescent probes to detecting hydrogen peroxide and hydrazine in living cells.Currently,most of the hydrogen peroxide fluorescent probes are tended to be interfered and show slow response.In our previous work we found that a fluorescent probe with 2-chlorophenyl-2-(azidomethyl)benzoate structure could rapidly respond to hydrogen peroxide.Herein,we report CFAN as a ratiometric fluorescent probe for hydrogen peroxide based on a FRET mechanism.This probe exhibited high selectivity and sensitivity in response to hydrogen peroxide in aqueous solution.What's more,the ratios exhibited an excellent linear relationship with the concentrations of hydrogen peroxide from 25 to 300 ?M,suggesting that the probe was potentially useful for quantitative determination of hydrogen peroxide.And the detection limit was calculated to be 0.5012 ?M.Furthermore,we assessed the application of the probe for monitoring and imaging of hydrogen peroxide in living cells.All of the results indicated that probe CFAN was cell membrane permeable and could be employed for ratiometric fluorescence imaging of hydrogen peroxide in living cells.We accidentally found that the intermediate CFAc can effectively detect hydrazine in aqueous solution.Then we successfully applied CFAc for sensing hydrazine in aqueous and living HeLa cells.This probe exhibited high selectivity and sensitivity in response to hydrazine in aqueous solution.Considering environmental and biological applications,the fluorescence signals of CFAc to hydrazine were investigated in CH3CN/PBS buffer,and a suitable ratio(1/7)was chosen after a careful investigations.What's more,the ratios exhibited an excellent linear relationship with the concentrations of hydrazine from 350 to 600 ?M,suggesting that the probe was potentially useful for quantitative determination of hydrazine.The detection limit was calculated to be 0.047 ?M.Further,it showed low cytotoxicity and was successfully applied to dual-channel imaging of hydrazine in living cells.We expect the new probes to be utilized in a variety of chemical and biological applications.In this paper,CFAN was reported as a ratiometric fluorescent probe for hydrogen peroxide based on a FRET mechanism.2-Chlorophenyl-2-(azidomethyl)benzoate structure was used as a new hydrogen peroxide detection site in the designing of CFAN.Accidentally,during the synthesis of CFAN,we developed a ratiometric fluorescent probe(CFAc)for hydrazine based on FRET mechanism for the first time.The probes were characterized by HRMS and NMR,and then studied for their spectral properties.We have developed two novel fluorescent probe with fast response and high selectivity under physiological conditions.Preliminary confocal laser scanning microscopy experiments show that probes have potential for practical application in living cells.