Enzymatic - Fluorescence Quenching Method In Drug Analysis Applications

The quantity of the druggery affects people's health and safety directly. Therefore the whole control of druggery plays an important role in the clinic and medical treatment.Spectrophotometry, chromatography and spectrophotometry- chromatography, as the most essential methods in the druggery measurements, have been widely used in the determination of pharmic products. In recent years, fluorimetry is developing in the study of analysis and druggery measurements, due to its highly sensitivity and good selectivity, simple procedure, less sampling and easy control apparatus. Obviously, fluorimetry is one of the methods in the quatitative determination of drugs in the modern instrumental analysis.To improve the sensitivity and selectivity, in this paper, using molecule spectroscopy (fluorescence and UV spectroscopy) as research instrument, some novel and simple methods for the assay of drugs have been established in my master paper.There are five chapters in this paper, we have determined Dopamine,Adrenalin and Procaine hydrochloride.The first part of this paper reviews the application and prospect of derivative spectrophotometry in drug analysis in recent years.The second part, we selected HRP as enzyme, we studied the interaction between complex and dopamine. The tyrosine reacts with H2O2 to form product which is a strong fluorescence substance in the presence of HRP, The fluorescence intensity of product decreased in the presence of dopamine, which showed fluorescence quenching occurred. A new, selective and sensitive method for determining dopamine has been established based on the principle of fluorescence quenching. This method is can be used for the determination of adrenalin and vancomycin.The third part, Because of the interference of background of acetylacetone formaldehyde, some primary amine-containing compounds cannot be determined by normal fluorimetry using acetylacetone-formaldehyde as the fluorigenic reagent. The procaine hydrochloride was studied as a sample in the synchronous fluorimetry using acetylacetone-formaldehyde as the fluorigenic reagent and the background of acetylacetone-formaldehyde was eliminated without separation. Some primary amine-containing compounds, such as procaine hydrochloride, L-phenylalanine, n-propylamine, n-butylamine, L-Glutamic acid, DL-trytophan and L-leucine can be determined by synchronous fluorimetry. The linear regression equations of calibration graph, detection limits, recoveries and optimum condition were listed.The fourth part, it is fundamental that the absorbance-difference method is developed for determination of procaine hydrochloride based on the measurement of changes in the absorbance differences (?A) between the absorbance of two wavelengths. The results presented in this paper demonstrated clearly that absorbance difference at the two wavelengths is proportion to the analyte concentration, and this method can eliminate the interference caused by background and stray radiation. The results obtained agreed with those of the official method (dead-stop titration). In addition, the method is rapid and simple.