Ds-simultaneous Study Multi-component Chinese Herbal Medicine With Bovine Serum Albumin

As abundant protein in blood plasma, serum albumin can widely combine with endogenous and exogenous substances, and the study on the interaction of small molecular drugs with serum albumin are very active. Recently, basic research between natural drug molecules and biomacromolecule has gradually become the focus of attention for domestic and foreign scholars. Chinese herb is regarded as a treasure of China, the research about interaction between active pharmaceutical ingredients of it and biomacromolecule was rather late. Moreover, the interaction mechanism on small molecule drugs and biomacromolecule was lack of deep-seated understanding. Therefore, the research on interaction mechanism of active components of Chinese herb and biomacromolecule has become one of the important subjects on interdisciplinary development of chemistry, life science and medicine.Spectrometry, as a common method of interaction between drug molecules and biomacromolecule, can't be high-throughput study on the interaction.However, the chemical composition of Chinese herb was very complex. If the active components of Chinese herb are separated, and then mechanism was studied one by one, it would be on the contrary to the principles of integrity and coordination of Chinese herb. Therefore, the system that online dialysis sampling combine with HPLC was established in this thesis. The system was used in the research on interactions of Salvia salvia water-soluble components and bovine serum albumin, and problem that the study on interactions between a variety of components in Chinese herb and biomacromolecule at the same time was solved too based on the system. It can provide information and basis for the research on the interaction between biomacromolecule and active components of Chinese herb.This paper includes three chapters.Chapter one:Review.Firstly, from protein structure, the structure of serum albumin and optical properties of serum albumin, description about the biological basis of research on interaction of molecule and biomacromolecule was given; And then, comments that research on interaction of small molecule drugs and serum albumin and theory, application and problems et cetera of several commonly used methods was reviewed. And identification and mechanism of small molecules and serum albumin was deeply studied theoretically; Finally, the establishment of on-line dialysis and HPLC method and application of the method on research of interaction between a variety of components in Chinese herb and biomacromolecule was regarded.Chapter two:The establishment of On-line dialysis sampling-High Performance Liquid Chromatography system and study on dialysis characteristics of water-soluble components in salvia.Based on the principles of integrity and coordination in Chinese herb treatment, the system that online dialysis sampling combine with high performance liquid chromatography was established in this thesis, and research on binding mechanism of a variety of components in Chinese herb and serum albumin was carried out. Water-soluble components in salvia,as a carrier in this chapter, was studied more comprehensive on dialysis characteristics and HPLC detection.Through experiment we discussed influence on the dialysis rate of Chinese herb molecules by the factors of dialysis membrane area, perfusion flow rate, the final choice of the optimal conditions were:perfusion flow rate of 2.5μL/min and micro-dialysis membrane length of 9 cm.Experimental results show that the system has good accuracy and reliability. At the same time, qualitative analysis on danshen injection components by LC-MS was conducted, and danshensu, protocatechuic acid, protocatechuic aldehyde, caffeic acid, ferulic acid and other substances were identified.Chapter three:Study on binding characteristics on water-soluble components and bovine serum albumin in danshen by DS-HPLC system.In this paper, research on binding mechanism on water-soluble components in danshen and bovine serum albumin was carried out by DS-HPLC system under the conditions that pH 7.4 close to physiological environment in PBS buffer. From the perspective of chromatography and ultraviolet spectroscopy, experiment do research on the influence that degree of drug combination by buffer pH and temperature, and degree of drug molecules, binding constants and thermodynamic constants were obtained. Furthermore, according to different thermodynamic constants, the interaction force between danshensu, protocatechuic acid, protocatechuic aldehyde, caffeic acid, ferulic acid and bovine serum albumin was determined.