Adsorption Isotherm Of Non-linear Chromatography And Enzyme-Catalyzed Reaction Chromatography

Adsorption isotherm is the fundamental of nonlinear chromatography theory. The instruction sense of it lies in when the adsorption isotherms of preparation components behave nonlinear, the characteristic and rules of chromatography system in time and space might be acquired under a certain case by nonlinear chromatography. So the procedure of preparation chromatography would be optimized.In this dissertation, method of determination of binary system isotherms was applied to ginsenoside Rg1 and Re adsorbed in C18. And the isotherms of ginsenoside Rg1 and Re were measured. The retention time of slow component ginsenoside Re was predicted with the results of non-linear chromatography theory. The isotherm constants G1, G2 and coefficients b1, b2 of gensenoside Rg1 and Re could apply an optimum condition for further separation gensenoside Rg1 and Re by preparative chromatography. And also G1, G2 and b1, b2 could get ready for the course of separation gensenoside Rg1 and Re by means of enzymatic catalyzed reaction chromatography.The competitive adsorbed amount of 2-phenylethanol and 3-phenylpropanol were measured by binary frontal analysis under the case of relative higher concentration. Three mathematical models, i.e. competitive Langmuir, general competitive Freundlich and multi-component Langmuir Freundlich were applied to fit the experimental data. The competitive general Freundlich model fit very well for different concentration ratio in the whole concentration range. The model improved effectively the problem of less precision caused by competitive Langmuir model.Reaction chromatography gains a lot of concern as a kind of technology of high performance separation and preparation. In this dissertation, the systems of glucose isomerization reaction chromatography and maltose enzyme catalyzed reaction chromatography served as the exploration experiments with NH2 binded packing material served as the stationary phase. Glucose isomerization and maltose catalyzed conversion realized in an analytical column, respectively. The elutions of glucose isomerization reaction chromatography and maltose catalyzed reaction chromatography were measured and Michaelis-Menten constant Km and Vm were acquired for both systems, respectively. Regarding of Michaelis-Menten constant Km and Vm, the applicability of the analysis solution of non-linear reaction chromatography was explained and discussed. The isotherm coefficient b and reactant rate kr of the reactant glucose and maltose were measured respectively by means of non-linear reaction chromatography analysis solution under the case of enzyme anticipated reaction. Compared with either chromatography elution method or dynamic Michaelis-Menten method, kr for both reaction chromatography systems were fitted very well. And the important significance of applying non-linear reaction chromatography has shown that isotherm coefficient b might be measured under the case of enzyme anticipated in the reaction.Under the case of preparation, the procedure of reaction chromatography of enzyme catalyzed starch by a-amylase has been carried out with C18 as the stationary to investigate the operative conditions (such as temperature, pH, concentration of reactant and concentration of the enzyme) and the system conditions (diameters of column and particle size of packing material). The intermediate product dextrin hydrate was acquired. The results showed that C18 is capable of separation for starch and dextrin hydrate. Factors of different size of packing material (20μm and 50μm), shape of packing material (spherical and irregular) and column size (1×10 cm,2×10 cm) were evaluated on the aspect of the extent of realization reaction chromatography.In this dissertation, enzyme catalyzed reaction chromatography in terms of A(?)B type has been investigated. The results in this dissertation have an important significance for isomerization or chiral separation system. Also such results would be an experimental or technical support for further development of continuous reaction chromatography technique such as simulated moving bed reactor (SMBR).