Synthesis And Application Of Porphyrin Fluorescent Probes For The Detection Of Several Bioactive Substances

Fluorescence method,attracted great attention of researchers,compared with other methods,has many advantages such as high sensitivity and short response time in the detection of biological substances.Bioactive substances?mercapto compounds?Cys,Hcy and GSH?,methylglyoxal,nitric oxide,etc.?play a key role in many biological processes.Therefore,the design and synthesis of fluorescence probes with excellent performance,owing to the short transmission wavelength,single signal,poor biocompatibility and other deficiencies,are of great significance for the detection of biothiol,methyl glyoxal and nitric oxide molecules,which are mainly discussed from the following three aspects:?1?Synthesized and designed two fluorescence porphyrin probes of?4-?20-?4-??7-nitrobenzo[c][1,2,5]oxadiazol-4-yl?oxy?phenyl?-10,15-diphenylporphyrin-5-yl?phenol?Tz1?and 7,7'-???10,15-diphenylporphyrin-5,20-diyl?bis?4,1-phenylene??bis?oxy??bis?4-nitrobenzo[c][1,2,5]oxadiazole??Tz2??,which choose porphyrin as materials to simultaneously detect three mercapto compounds?Cys,Hcy and GSH?.The structure of the target compound was confirmed by mass spectrometry and nuclear magnetic resonance hydrogen spectrometry.The near-infrared region?>600 nm?provides visual information in deeper tissues,reduces autofluorescence?background?,improves sensitivity,and reduces light damage and bleaching.Ratio fluorescent probes have two different wavelength signals and can provide an internal standard to eliminate interference.Due to the poor solubility of Tz2,and the fluorescence response is not very different from that of Tz1.Therefore,the test results of Tz1 on three mercapto compounds are mainly used in this paper to illustrate.Firstly,the optimal reaction conditions for the detection of three mercapto compounds by the probe?Tz1?were determined through condition optimization,such as the optimal solvent,buffer solution,pH value and response time.The effect of the concentrations of three mercapto compounds on fluorescence intensity was investigated.The results show that the relationship of Tz1 and Cys and Hcy or GSH,the concentration of Cys is ranging from 0.012 to 10?mol?L^-1;Hcy is ranging from 0.020 to 10?mol?L^-1;GSH is ranging from 0.16 to 20?mol?L^-1,have a good linear.The results of the selective and anti-interference tests showed that the presence of other interfering substances would not interfere with the detection of the three mercapto compounds.Meanwhile,the application of Tz1 to detect the content of three sulfhydryl compounds in normal human serum was explored.?2?A porphyrin fluorescence probe?5-?3,4-diaminophenyl?-10,15,20-triphenyl porphyrin?Tz3??with single wavelength response and near infrared detection of methylglyoxal was designed and synthesized from porphyrin.The structure of the target compound was confirmed by mass spectrometry and nuclear magnetic resonance hydrogen spectrometry.Firstly,the ultraviolet and fluorescence spectra of methylglyoxal detected by the probe were obtained,and the optimal conditions for the detection of methylglyoxal by the probe were selected.The effects of different concentrations of methylglyoxal on fluorescence intensity were investigated.The results show that the relationship of Tz3 and MGO,the concentration of MGO is ranging from 0.0096 to 20?mol?L^-1,has a good linear,the detection of limit is 9.6 nmol?L^-1?3?/k?.The results of selective and anti-interference experiments show that the probe has good anti-interference ability and selectivity for the detection of methylglyoxal.Meanwhile,Tz3 was also applied to the detection and imaging of methylglyoxal in HeLa cells.?3?A ratio fluorescent probe?5-?3,4-diaminophenyl?-10,15,20-triphenyl porphyrin-PEO-PS-AEMH-EANI?Tz4??with dual functions of near infrared emission and water solubility was prepared by ultrasonic method using diaminophenyl porphyrin as fluorescence signal reporter group and polymer as carrier.The reaction unit of the probe?Tz4?with the structure of o-phenylenediamine as the recognition site of nitric oxide,porphyrins act as near-infrared fluorophore,and the polymer provides a good water-soluble environment for porphyrins.Probe?Tz4?can detect nitric oxide in aqueous solution.Firstly,the optimal conditions for the detection of NO by Tz4 was selected.At the same time,the influence of nitric oxide concentration on the reaction is studied.The results show that the relationship of Tz4 and NO,the concentration of Cys is ranging from 0.0242 to 30?mol?L^-1,has a good linear,the detection of limit is 24.2 nmol?L^-1?3?/k?.The results of selectivity and anti-interference test showed that Tz4 had good selectivity and anti-interference ability in the detection of nitric oxide in aqueous solution.Finally,it will have the potential ability to image nitric oxide in cells.