Studies On The Determination Of Dopamine And Glutathione Based On Reactive Fluorescence Method

With the development of life science, the important role in life science ofbiological small molecules, such as dopamine (DA) and glutathione (GSH), werewidely recognized by people. Therefore, establishing rapid, accurate and convenienttesting method of dopamine and glutathione on life sciences, clinical medicine, andother fields has important significance. Due to the high sensitivity, good selectivity, theadvantages of simple and good stability, fluorescence analysis method has been widelyapplied in the detection of biological molecules. In this thesis, two kinds of dopamineand three kinds of glutathione determination methods were proposed based onspecificity of fluorescent reaction between dopamine, glutathione and fluorescentreagent.This thesis mainly divided into the following five parts:(1) A fluorescent strategy for dopamine (DA) detection is proposed using the2,3-diaminophenazine (DAP) as the signal readout, as DAP display a relatively strongfluorescence emission. The aminochrome, formed by the autooxidation of dopaminewith manganese as Catalyst, can condense with DAP, leading to the fluorescencequenching of DAP at a certain concentration. The concentration of dopamine can bedetermined by fluorescence spectrophotometer. Thus, a linear range of2.0×10-6Mto6.1×10-5M with a detection limit of1.76×10-6M (3σ, n=11) for DA wasobtained.(2) Using glyoxal as the fluorescent derivatives reagent to react with DA, thefluorescence detection of DA was proposal. Their derivative displays a strongfluorescence emission. The method has a good detection for DA with a linear rangefrom1.0×10-7to1.0×10-5M, the detection limit is3.35×10-8M.(3) The reaction of GSH with Cu2+and the effect of GSH on the fluorescent systems, i.e. alizarin red (ARS)-H3BO3-Cu2+were studied. Cu2+can quench thefluorescence of ARS-H3BO3. With addition of GSH on which the mercapto (-SH) hasa firmer combination with Cu2+, GSH would wrested the Cu2+which was combinedwith ARS, resulting in the recovery of fluorescence. As a result, GSH was detectedrapidly, with the linear range from1.0×10-5to1.9×10-4M and the detection limit of1.0×10-5M.(4)1,8-anthracene disulfonyl methylamine display strong fluorescenceemission. Hg2+could quench the fluorescence, the mercapto (-SH) of GSH addedinto the system had a firmer combination with Hg2+, resulting in the recovery offluorescence. The GSH could be quantified in the ranges of2.5×10-6to3.5×10-5Mand the detection limit is1.68×10-6M.(5)5-(2-Methyl-4-trifluoromethyl-thiazol-5-yl)-[1,3,4] thiadiazol-2-ylamine(MTA) with strong fluorescence emission was synthesized. Complexation betweenMTA and Hg2+result in fluorescence quenching. The mercapto (-SH) of GSH hasstrong complexing ability with Hg2+, resulting in the recovery of fluorescence. theGSH could be detected in the ranges of1.0×10-5～5.0×10-4M and the detection limitis6.0×10-6M.