Investigation Of Binding Ratios Between Sudan And Human Plasma Proteins By Cloud-Point Extraction And Separation Of Proteins

Investigating the interaction between small molecules and plasma proteins is very important in pharmacokinetics and toxicology. The changes of binding ratios between small molecules and plasma proteins affected by varied factors in the body would result in the corresponding changes of dynamic parameters of metabolism. The binding between small molecules and plasma proteins would influence the distribution, metabolism and excretion of small molecules and further their binding abilities with their receptor. Especially, the researches about the binding between drugs and proteins would provide an important theoretical reliability on studying the causes of human disease, the revolution of dosage forms and development of new drugs. In this dissertation, sudan, a class of organic dyes, interacting with human plasma proteins was studied. On the basis of the previous researches, the following major innovative works were carried out:(1) Sudan was chosen to study its binding with plasma proteins in vitro. The binding ratios of single sudan or sudan mixture and plasma proteins were compared. The results indicated that four sudan substances were not competitive with each other to act with plasma proteins.(2) A novel bifunctional SBA-15 was used as sorbent to separate proteins preliminarily. The results are satisfied.This dissertation consists of three chapters.Chapter 1:The nature, ingredients and functions of plasma proteins, especially, t the structure and function of Human Serum Albumin were briefly introduced. The studying methods, theories, and developments of binding between small molecules and plasma proteins and the separation techniques of proteins were summarized.Chapter 2:The binding ratios between sudan and plasma proteins by cloud point extraction and HPLC were studied. The binding ratios of mixture or single sudan were calculated, and the results were compared with those of equilibrium dialysis by T test. The comparable result indicated that there was no obvious difference between two methods.Chapter 3:Investigating the selective adsorption and desorption of proteins which have various pI and abundance is carried by a novel micro solid extraction system of pipette tip loaded with the original and bifunctional SBA-15 coupled with different pore sizes. The results indicated that this novel micro solid extraction system of pipette tip can be successfully used to separate the high abundance proteins and enrich low abundance proteins by modifying pH of the solution and the volume ratio of buffer and organic reagent in the eluent to make sure that the signals of high-abundance proteins could not bury those of low-abundance proteins and the detection of low proteins can be carried easily.