The Establishment Of Fluorescence Analysis Method And The Application In Biologies

This experiment analysed the principle and characteristic of fluorescence analysis,and itsapplication of content measurement in pharmaceutical and nitrite residue in vegetables, whichincluded fluorescence analysis of vitaminB₂and catalytic fluorescence analysis of nitrite.Firstly, we measured the excitation and emission wavelength of vitaminB₂, determinedand chose the optimal wavelength. Secondly, we researched the experiment condition ofvitaminB₂by fluorescence analysis, and simplified precondition process, reduced the dosageof experiment reactants. Finally, we gained the convenient shortcut method, ideal analyticresult and higher accuracy;Through orthogonal test, we determined the sequence of impactingfactors on vitamin B₂, which demonstrated that determination time in solution system was thelargest impact factor; We determined the optimal determination time, temperature and pH.Under the condition of λex=490.0nm and λem=512.0nm, recovery rate of vitaminB₂was96.58～100.37%and variation coefficient was0.10%. This paper adapt orthogonal test anddrew the optimal determinate conclusion: solution set time was45min, measure temperaturewas12℃and light intensity was dark light. The impact sequence of the three factors onvitamin B₂was measure time> temperature> light intensity.Nitrite catalyzed neutral red fluorescence bleaching reaction. Nitrite concentration andreaction rate was proportional to fade. Through single factor analysed the choice of variousexperimental conditions, and optimized the use of uniform experimental conditions, wedetermined the best combination of factors. Through determinated nitrite residues of plantstem of potatoes and onions, we can draw the conclusion that the skin of vegetables alsocontain sulfite residues, but the content was not very great. Vegetables used in thedetermination of residual nitrite by this method, at the excitation wavelength λex=537.0nmand emission wavelength λem=595.0nm,and variation coefficient was6.5%, the averagerecovery can be achieved100.2%...