Studying And Application Of Resonance Light Scattering Technique In Protein Analysis

Protein is the most important parts and the most functional microorganism of life, the life origin, the existence and the evolution all makes closer connection with the proteins. Chemist and biologist have always focused on the study of proteins analysis, the protein determination has relation to the biochemistry and clinic analyse. With the rapid development of life sciences, biological engineering and environmental science, at the same time analysis science is facing new opportunity and challenge. As the analysis of the complexity of the object, the trace and ultra-trace components of the complex matrix to separate and detect become a key problem. Therefore, in this research paper, in order to analysis the trace components of the protein and drugs, RLS methods were established to the determination of these biological macromolecules of detecting protein using resonance light scattering, and a new molecularly imprinted-chemiluminescence method for the determination of ascorbic acid was developed. Their experimental conditions have been investigated systematically. The more detailed novelty of this research can be categorized as following:1. The resonance light-scattering (RLS) spectra and absorption spectra of Ery-Cu(II) metal complex with bovine serum albumin (BSA) were studied. In the Brition-Robinson buffer at pH=3.78, the RLS of Ery-Cu(II)-protein system can be greatly enhanced at338nm. Under optimal conditions, the linear range of calibration was0.5～10μg/mL, the linear regression equation was IRLS=1.7146c+16.327(BSA, μg/mL) with r=0.9981, and a detection limits was0.16μg/mL. The developed method was applied to the determination of BSA in synthetic samples with satisfactory results. A mechanism of electrostatic force was suggested for the interaction between Ery-Cu(II) complex and BSA.2. A new Rayleigh light scattering (RLS) assay of protein was conducted. At the optimum pH conditions, and in the presence of Cu(II), the weak RLS of ACBK can be enhanced greatly by the addition of proteina. Based on this, the reactions of ACBK-Cu(II) and proteins were studied. A new quantitative determination method for proteins has been developed. The method is simple, practical, and relatively free interference from coexisting substances, as well as much more sencitive (the dynamic ranges of0.2～6.0μg/mL and detection limit of0.12μg/mL for bovine serum albumin (BSA)) than most of the existing assays.3. Molecularly imprinted polymer was prepared by step-polymerization reaction of epoxy resin using ascorbic acid as the template molecule, diethylenetriamine as curing agent and polyethylene glycol as pore-forming reagent. A new chemiluminescence method for the determination of vitamin C was proposed by the combination of ascorbic acid-imprinted polymer and KMnO4-formaldehyde-ascorbic acid-CL system. The linear range is8.0×10-6～6.0×10-3mol/L, the detection limit is5×10-4mol/L, the relative standard deviation is2.7%(n=11). This method has been successfully applied to the determination of vitamin C in dury samples with satisfactory results.