| Resonance Rayleigh scattering (RRS) and resonance nonlinear scattering (RNLS) such as double frequency scattering and second order scattering etc. is a new technology developed in 1990s. For its remarkable characteristics of high sensitivity, simple operation and good selectivity, this method brings to more attention and interesting and has been studied and applied more and more in recent years. Particularly, it is very sensitive to the nonbonding action of biological macromolecules, therefore, has been used as a useful means to study biological macromolecules. At present, this technology is mainly used to the study of nucleic acids and proteins. However, up to now it is seldom applied to the study of another most important biological molecule, saccharide. Moreover, There is no report about this technology used to the determination of some physicochemical parameters. Therefore, supported by the National Natural Science Foundation of China and using heparin, a glycosaminoglycan, as an example, this thesis has studied the application of the RRS and RNLS methods. Furthermore, using the inclusion constant of p-cyclodextrin and the critical micelle concentration (CMC) of some surfactants as examples, the methods have been applied to the determination of the physicochemical parameters.The main contents and some conclusions of the thesis are as follows:1. Resonance Rayleigh Scattering Study of Interaction of Heparin with Some Basic Diphenyl Naphthylmethane Dyes and Some Basic Phenothiazine Dyes and Their Analytical ApplicationUsing heparin as an analyte, the RRS spectral characteristics of the interaction of heparin with some basic diphenyl naphthylmethane dyes and some basic phenothiazine dyes and their analytical application have been studied. The results show that in weak acidic and weak basic medium, binding of heparin with some basic diphenylnaphthylmethane dyes such as victoria blue 4R (VB4R), victoria blue B (VBB) or night blue (NB) and some basic phenothiazine dyes such as methylene blue (MB), Azure A Azure B or toluidine blue (TB) can result in a significant enhancement of resonance Rayleigh scattering (RRS) and appearance of a new RRS spectrum. Their maximum scattering wavelengths max appear at 523 run, 534 nm, 540 nm, 346 nm, 337 nm, 338 run and 329 nm for VB4R, VBB, NB, MB, Azure A, Azure B and TB, respectively. The RRS spectral characteristics, the influencing factors and the optimum conditions of these reactions have been investigated. By combining ultra violet visible spectrophotometry and the quantum chemistry calculation, the reaction mechanism has been discussed. The results show that the RRS spectral characteristics of the combination products of heparin with these basic dyes are closely related to their absorption spectra. There is a corresponding relationship between them. And the relationship between the RRS intensity and the concentration of heparin has been investigated. A new method for the determination of trace amounts of heparin based on the RRS method has been developed. The methods exhibit high sensitivities and the detection limits for heparin of the seven systems are between 3.35 ng/mL and 35.93 ng/mL. Among them the VB4R system is the most sensitive and was applied to the determination of heparin in heparin sodium injection samples with satisfactory results. It also illustrates that this technology has a good application foreground to the study and determination of saccharide. This is new development of the RRS technology used in the analytical application of the biological macromolecules.2. Study on Frequency Doubling Scattering and Second-Order Scattering Spectra of Heparin-Methylene Blue SystemThe combination of heparin with methylene blue, a basic phenothiazine dye, has been studied by means of the frequency doubling scattering and second-order scattering technology. The results show that the binding of heparin with methylene blue can result in a significant enhancement of frequency doubling scattering (FDS) and second-order scattering (SOS). Their maximum scatte...
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