Nanoseparation And Characterization Of Gold Nanoparticles And Their Effects On2D6Activity In Drug-metabolizing Cytochrome P450Enzyme Systems

Gold nanoparticles have been widely used as one of the most versatile nanodevices in a variety of fields such as sensors, biomedicine fields and so on, due to the unique physico-chemical properties. It is of great concern that they may also cause adverse effects on the human body with the increasing of human and environment exposure even making progress in society at the same time. Cytochrome P450enzymes located in liver plays an important part in the process of chemical alteration of a variety of endogenous and xenobiotic compounds. Therefore, the interaction research of gold nanoparticles with the cytochrome P450enzyme could have a biologically significance on safety problems of gold nanoparticles, using in vitro metabolism system. Water-miscible organic solutions is the most convenient system, for the sake of direct measurement of organic (bio-) molecules in complex samples and decrease matrix effects, the pretreatment method of gold nanoparticles serves as a link between past and future experiment for reasons such as noted above. On the other hand, capillary electrophoresis is the most efficient and convenient detection means to analyze biomacromolecule and nano-particle at present. For these reasons, developed and validated a simple and rapid phase transition way for reject gold nanoparticles based on the phenomenon that the acetonitrile can be separated from water by adding inorganic salt; based on the above method, preliminary investigate concentration and size effect of gold nanoparticles on the activity of cytochrome P4502D6in vitro; investigate the application of capillary electrophoresis in cytochrome P450and gold nanoparticles to explore a methodology attempt between cytochrome P450and gold nanoparticles. In this thesis, the main work is as follows:1. Based on the phenomenon that the acetonitrile can be separated from water by adding inorganic salt, developed a simple and rapid phase transition way for reject gold nanoparticles. In this study, the salt-induced effects of different amounts of salt additives, including magnesium sulfate (MgSO4), sodium sulfate (Na2SO4), magnesium chloride (MgCl2) and sodium chloride (NaCl), on the rejection of gold nanoparticles with different diameters ranging from5to100nm were compared. Incubation time and salt addition also have been investigated. Characterization of the particles before and after phase separation was performed by using UV-Vis spectroscopy, digital camera imaging and transmission electron microscopy. This approach also has been applied to determination dextromethorphan and its metabolite in human plasma and liver enzyme. The result of the experiment was satisfactory.2. Based on the above method, using dextromethorphan as substrate probe:compare a series of concentration of5,7,10,20,50,100nm gold nanoparticles effect on the activity of cytochrome P4502D6in vitro. The results of this study is to show that gold nanoparticles have concentration and size effect, which is a valuable significance to interaction research of gold nanoparticles with the cytochrome P450enzyme and toxicity study of gold nanoparticles.3. Based on capillary electrophoresis, a capillary zone electrophoresis method have been developed for the rapid determination of phenacetin, acetaminophen, dextromethorphan, dextrophan; investigate field amplified sample injection on-line preconcentration effect on the sensitivity of capillary zone electrophoresis; investigate electrophoretic of gold nanoparticles in capillary to obtain potential methodology for interaction research of gold nanoparticles with the cytochrome P450enzyme.