Novel Rayleigh Light Scattering Methods For Quantitative Analysis Of Protein

Rayleigh light scattering (RLS) is a new technology developed in 1990s for trace analysis. It was founded by Pasternalk et al in 1993. There are more researches and applications of RLS recently. It is widely used in the research and determination of proteins, nucleic acids, carbohydrates, medication, non-particle, inorganic particles and surfactants and shows great foreground of use. Comparing with other methods, the RLS method has more acuminous sensitivity, more simple manipulation. And it is able to provide details for molecule's configuration. However, the RLS method has disadvantage in reproducibility and stability.In this dissertation, on the basis of the previous research, novel Rayleigh light scattering methods were developed to investigate the quantification of proteins in human blood serum. The following major innovative works were carried out:1: The flow injection combined with Rayleigh light scattering technique was used to quantify the protein in human serum samples. The homemade instruments of flow injection were connected with spectrofluorophotometer. Protein in human serum samples were determined using the combination technique mentioned above, the results' reproducibility and stability were elevated.2: A novel system of OP-HSA- Brilliant ponceau 5R was founded for the quantitative detection of protein in human serum samples by Rayleigh light scattering. The method had getten more acuminous sensitivity.This dissertation consists of three chapters: Chapter 1: The principles, up to date applications and novel development of Rayleigh light scattering were briefly described. And the surfactants used in this dissertation were mentioned.Chapter 2: A novel flow injection analysis (FIA) method coupled with Rayleigh light scattering (RLS) was developed for the determination of protein concentrations. This method is based on that the weak intensity of RLS signals of dodecane-1-sulfonic acid sodium salt can be enhanced by the addition of protein in weakly acidic solution. The determination limit was 70ng/mL for human serum albumin, the linear range was 3.0-28.0μg/mL, the maximum relative standard deviations (R.S.D.) was no more than 2.12% and the recovery of the samples was between 97.83-106.09%. The FIA-RLS method was more stabile, rapid, and reproducible than the general RLS method.Chapter 3: In the presence of OP and in the pH2.11 buffer, the Brilliant ponceau 5R combined proteins to form complex, causing an enhance of RLS signals. The sensitivity raised 2 times in the presence of OP. For human serum albumin, the linear range was 0-10.0μg/mL, and the determination limit was 5ng/mL. The recovery of the samples was between 97.80-109.62%. This method was more sensitive than the general RLS method and obtained satisfactory results.