| Scientific instruments lay foundation to promoting the development of science and technology,and industrial upgrading,and their innovative research and performance improvement are of great significance to maintaining the healthy and rapid development of China’s economy and society.As a large-scale separation and purification equipment towards the active components of natural products,synthetic compounds and other substances,high-speed counter current chromatography(HSCCC)instruments characterize no dead adsorption,no solid support,good tolerance,flexible elution mode,high recovery rate and good reproducibility.Thus,tehy can greatly reduce the economic cost,environmental pollution and resource waste in pharmaceutical industry,which plays a key role in promoting the rapid development of China’s biopharmaceutical industry.However,due to the problems of current HSCCC instruments including low column efficiency and unknown factors affecting column efficiency,etc,they are not able to meet the demand for the separation of complex active substances such as chiral compounds and proteins.This dissertation originates from a special subject of the National Natural Science Foundation of China―development of major scientific instruments,and is entitled“the development and application of high-efficiency HSCCC instrument”.In order to solve the problems of the low column efficiency in current HSCCC instruments,this project has carried out extensive research on the multifunctional experimental device of HSCCC,influencing factors of column efficiency,optimization technology of column efficiency and high-performance prototype,with combining medical and engineering talents.The main research work and results are as follows:(1)Development of experimental instrument.Based on the analysis of the mechanism of two-phase unidirectional hydrodynamic dynamic balance and its mechanism realization principle,the structural design methods of multi-purpose frame,box planetary mechanism,variable planetary mechanism with changeableβvalues(the ratio of the rotation radius R to the revolution radius r of the column),multi-type columns and anti-twisting tubes were studied.A touchable interactive control system of HSCCC and a multi-functional HSCCC experimental instruments have been successfully developed,which can provide an experimental platform for the subsequent research on the factors influencing the column efficiency.(2)Research on the modeling method of sample separation process and the influence of the injection and detection links on the column effectiveness.In order to model the process of injection and elution,a continuous injection-elution model associated with injection process and elution process was established by transforming the continuous injection elution process into the gradual injection elution mode.At the same time,based on the assumption of segmented mass transfer,a micro-segmented sample transfer model suitable for the detection process was established.By combining the ctwo models,a sample elution-transfer model for the separation process of HSCCC instrument was established,which realized the simulation of peak deformation caused by injection and axial diffusion caused by detection.Based on the simulation analysis of the model and the comparative analysis of the experiment,the modeling-method rationality of the sample elution-transfer model was verified,and the influence mechanism and magnitude of the column efficiency in the sampling and detection links were clarified,which laid a theoretical foundation for the independent analysis of the influencing factors of the column efficiency in the elution link,and also provided a theoretical basis for the optimization design of the injection-detection loop.(3)Research on visualization experiment and the influencing factors of column efficiency in elution process.To clarify the mechanism of column efficiency in the sample elution process,a visual experimental system was built based on the multifunctional experimental platform,and the two-phase dynamic characteristics of the sample elution process were analyzed.Through the analysis of the correlation between the two-phase dynamic characteristics and separation results,the elution model of incomplete mixing cell was established.Then,the elution transfer model was optimized,which realized the mathematical correlation between the instrument operation parameters,two-phase dynamic characteristic parameters and the whole separation process.Based on the simulation analysis of the model and the comparative analysis of the experiment,the effects of the frequency of two-phase periodic mixing and stratification,rotational speed,flow rate,mixing degree of the mobile phase and axial diffusion degree of the sample on the column efficiency were discussed,and the key factors affecting the column efficiency in the elution process were identified,that is,mixing degree of the mobile phase,which pointed out the direction for the improvement of HSCCC instrument design.(4)Optimization technology of column efficiency towards HSCCC instrument.According to the analysis of the key influencing factor of column efficiency,from the perspective of optimizing the main normal and binormal mixing degree of mobile phase,the column efficiency optimization methods of reducing average value ofβand increasing the inclination of annular groove of column were proposed respectively.First,based on the comparison of the two-phase force state of the helix tube under different r and R,the applicable conditions and feasibility of the optimization methods were demonstrated,and its experimental verification was carried out by using the multi-functional experimental platforms.For the second method,by constructing the movement model of the column with inclined annular groove,the adjustment mechanism of its two-phase binormal force was discussed,and the spatial motion trajectory equation at any point was derived,so as to determine the reasonable range of the inclination angle of the annular groove.In order to verify the feasibility of the theoretical analysis,the columns with inclination angles of 0~°and 8~°were manufactured,and the experiments of stationary phase retention and sample separation were carried out under various conditions.Experimental research showed that the two optimization methods of column efficiency can effectively improve the separation performance of HSCCC instrument,which provided technical support for the optimization design of the instrument.(5)Improved design and prototype development of analytical and semi-preparative HSCCC instruments.In order to solve the problem of low column efficiency of HSCCC instrument,based on the above research results,an improved design of analytical HSCCC instrument was carried out by using the anti-twisting technology of metal frame,and then an analytical HSCCC instrument with two side supports was developed,whose g level can be as high as 400g.At the same time,based on the design technology of analytical prototype,using the design method of large capacity column and the sealing technology that can realize reliable transfer from square tube to round tube,a semi-preparative HSCCC prototype with rectangular helix tube was developed.The experiments of stationary phase retention and sample separation under different working conditions showed that compared with traditional instruments of the same specification,the improved design of analytical HSCCC instrument has higher sample resolution and faster separation speed,and the improved design of semi-preparative HSCCC instrument has higher production efficiency,which laid the foundation for the localization of high-efficiency HSCCC instrument. |
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