Study On The New Application For The Determination Of Polysaccharose, Vitamin And Diuretics By Resonance Rayleigh Scattering And Fluorescence Spectroscopy

This paper mainly studies interaction of some medicine, such as chondroitin sulfate, vitamin B12, vitamin B1and torasemide with nile blue sulphate,12-tungstophosphoric acid and dihalogenated fluorescent dyes, and set up resonance Rayleigh scattering and fluorescence quenching methods for the determination of them. The methods have been successfully applied to the quick determination of chondroitin sulfate, vitamin B12, vitamin B1and torasemide in practical samples. The research contents are listed as follows:1. Determination of chondroitin sulfate by dual-wavelength overlapping resonance Rayleigh scattering method with nile blue sulphateIn a pH3.0～4.0Britton-Robinson (BR) buffer medium, CS and NBS could form the ion-association complex. As a result, the new spectra of resonance Rayleigh scattering (RRS), second-order scattering (SOS), and frequence doubling scattering (FDS) appeared and their intensities were enhanced greatly. The maximum wavelengths were located at303nm(RRS)、362nm(RRS),588nm(SOS) and350nm(FDS). And the three kinds of scattering intensities were proportional to the determination of CS in certain ranges. The methods had high sensitivity with the detection limit being3.4ng/mL for RRS (303nm) method,2.8ng/mL for RRS (362nm) method,7.1ng/mL for SOS method and FDS method. The characteristics of the spectra and optimal conditions of RRS method were investigated. The effect of coexistent substance on the determination of CS was studied, owing to the high sensitivity of RRS; it had been applied to the determination of CS in eye drops with satisfactory results. The recovery range was between99.4%and104.6%and the relative standard deviation (RSD) was between0.4%and0.8%.2. Study on the dual-wavelength overlapping resonance Rayleigh scattering spectra of the interactions of vitamin B12with12-tungstophosphoric acid and their analytical applicationsA new dual-wavelength overlapping resonance Rayleigh scattering (DWO-RRS) method was developed to detect vitamin B12(VB12) with12-tungstophosphoric acid (TP). In pH0-1.5HC1medium, vitamin B12(VB12) could react with12-tungstophosphoric acid (TP) to form a3:1ion-association complex. As a result, the new spectra of dual-wavelength overlapping resonance Rayleigh scattering (DWO-RRS), second order scattering (SOS) and frequence doubling scattering (FDS) appeared and their intensities were enhanced greatly. Their maximum wavelengths were located at330nm (RRS),370nm (RRS),608nm (SOS) and386nm (FDS), respectively. The three kinds of scattering intensities (△IRRS,△ISOS and△IFDS) were proportional to the concentration of VB12in certain ranges. The methods had high sensitivity with the detection limit being2.0ng/mL to7.6ng/mL. The optimal conditions and the effects of coexistent substances of RRS method were investigated. So, A simple, rapid and new method had been proposed to determine trace VB12. It had been applied to the determination of VB12in urine samples and vitamin B12tablets with satisfactory results. In addition, the reasons for RRS enhancement and composition of ion-association complex were discussed.3. Determination of vitamin B1with12-tungstophosphoric acid by resonance Rayleigh scattering methodIn HC1medium,12-tungstophosphoric acid (TP) reacted with vitamin B1to form a1:3ion-association complex. As a result, the new spectra of resonance Rayleigh scattering appeared and their intensities were enhanced greatly. The maximum wavelength was located at335nm. In the range of0.25～1.75μg/mL, the scattering intensities were proportional to the concentration of VB1. Based on this, the new method for the determination of VB1had been established. This method had high sensitivity and good selectivity, the detection limit (3a) was0.94ng/mL. It had been applied to the determination of VBi in urine samples and vitamin B1tablets with satisfactory result; the recovery was between97.3%-99.0%with RSD of1.1%～3.4%. The optimum reaction conditions, the RRS spectra characteristics and influencing factors had been discussed.4. Determination of torasemide by fluorescence quenching method with some dihalogenated fluorescent dyes as probesA novel fluorescence quenching method for the determination of torasemide (TOR) with some dihalogenated fluorescein dyes as fluorescence probes had been developed. In weak acid medium, TOR could interact with some dihalogenated fluorescein dyes such as dichlorofluorescein (DCF), dibromofluorescein (DBF) and diiodofluorescein (DIF) to form a binary complex, which led to fluorescence quenching of above dihalogenated fluorescein dyes. The maximum fluorescence emission wavelengths of them were located at532nm (DCF)、535nm (DBF) and554nm (DIF). The relative fluorescence intensity (△F=Fo-F) was proportional to the concentration of TOR in certain range. The detection limits were4.8ng/mL for TOR-DCF system,9.8ng/mL for TOR-DBF system and35.1ng/mL for TOR-DIF system. The optimum reaction conditions, influencing factors were studied; and the effect of coexisting substances was investigated owing to the highest sensitivity of TOR-DCF system. In addition, the reaction mechanism, composition and structure of the complex were discussed. The fluorescence quenching of dihalogenated fluorescein dyes by TOR was a static quenching process judging from the effect of temperature and the Stern-Volmer plots. The method was satisfactorily applied to the determination of TOR in tablets and human urine samples.