Synthesis And Application Of Luminol And Thiazole Orange Fluorescent Probes

1.Reaction conditions have been explored in this paper, and the synthetic route of luminol has been optimized. Chemiluminescenct system of H2O2-luminol catalyzed by blood was establised by fluorescence spectrophotometry through detecting the fluorescent value.Effects of buffer system, blood concentration and p H value on the chemiluminescent property of the system were studied to search the best luminescent conditions. Results obtained show that the chemiluminescent property of the system catalyzed by blood are best when using 3.0mmol/L Luminol and 56.60mmol/L Na2CO3-Na HCO3 as buffer system(solution A), mixing with 30.0mmol/L H2O2(solution B), p H value was 8.98.2.Phenolphthalin was prepared by reduction of Phenolphthalein using zinic power and glacial acetic acid. Potentiation of phenolphthalin for the luminescence of HRP-Luminol-H2O2 reaction system was explored. Result obtained indicates that good enhancing activities of phenolphthalin for the luminescence of HRP-Luminol-H2O2 reaction system are stable and emission intensity increases significantly, and the light emitting lasts more than 30 min. Mass concentration of HRP directly relates to chemiluminescent intensity of HRP-Luminol-H2O2 reactionsystem when HRP contents are 5-800 pg/m L, the equation of linear regression is lg I=1.07lgc+0.98, r=0.96, and a low detection limit of 1.25pg/m L HRP. RSD within-run is 2.8% ~ 5.1%(n=10), the recovery is93.5%~96.2%. Phenolphthalin can be used as quantity anaysis of HRP and its related markers.3.An improved method for the synthesis of fluorescent dye thiazole orange(TO-1) is described,the raw materials are commercially available diphenylamine and 2-mercaptobenzothiazole. 2-methylthio-N-methyl-benzothiazole p-toluenesulfonate is prepared through N-methylation and S-methylation of 2-mercaptobenzothiazole with Dimethylsulfate and methyl p-toluenesulfonate in 71.5% and 77.9% yield respectively.2-chloro-4-methyl-1-phenylquinolinium chloride was prepared by diphenylamine as starting materials,and thiazole orange was synthesized by the reaction of 2-methylthio-N-methylbenzothiazole p-toluenesulfonate and 2-chloro-4-methyl-1-phenylquinolinium chloride.