Enzymatic - Fluorescence Spectrometry In Drug Analysis Applications And Research

The quality of the druggery affects people's health and safety directly. Therefore the whole quality control of druggery plays an important role in the clinic and medical treatment.High performance liquid chromatography method, spectrophotometry method and electrochemistry method, as the most essential methods in the druggery measurements, have been widely used in the determination of pharmic products. In recent years, spectrophotofluorimetry is developing in the study of druggery measurements, due to its highly sensitivity, good selectivity, simple procedure and less sampling. Obviously, spectrophotofluorimetry is one of the methods in the quantitative determination of drugs in modern instrumental analysis.In this paper, using molecule fluorescence spectroscopy as research method, some novel and simple methods for the assay of drugs have been established. There are four chapters in this paper.The first chapter of this paper, the application and prospect of spectrophotometry, the development of horseradish peroxidase (HRP) and it's application, in drug analysis in recent years were discussed.The second chapter, we selected HRP as enzyme. Because of catalysis of HRP, the tyrosine reacted with H2O2 to form the product S which was a strong fluorescence substance. To the product S, the compounds (quercetin, troxerutin, curcumin) which called the flavonoids quenched the fluorescence intensity of product S. The reaction mechanism which was the static quenching process between product S and the flavonoids compounds. A new method for determining the flavonoids has been established based on the principle of fluorescence quenching. The fluorescence quenching mechanism was studied by the measurements of fluorescence lifetime and based on the Stern-Volmer plot.The third chapter, we studied the reaction between product S and berberine. The tyrosine reacts with H2O2 to form product S which is a strong fluorescence substance in the presence of HRP. The fluorescence intensity of product S decreased in the presence of berberine, which showed fluorescence quenching occurred. The reaction mechanism which was the static quenching process between product S and berberine. A new, selective and sensitive method for determining berberine has been established based on the principle of fluorescence quenching. The fluorescence quenching mechanism was studied by the measurements of fluorescence lifetime and the Stern-Volmer plot.The fourth chapter, a simple and convenient method was introduced for the determination of terramycin based on the principle of fluorescence quenching. An oxidation reaction of tyrosine with H2O2 catalyzed by horseradish peroxidase was studied, and product S was the oxidation product with a strong fluorescence. Terramycin quenched the fluorescence intensity of product S. The static quenching process was confirmed by the measurement of fluorescence lifetime on the basis of Stern-Volmer plot. The reaction mechanism which was the static quenching process between product S and terramycin. The enthalpy change, entropy change and free enery change were calculated.