The Development Of The Preparative Continuous Bed Chromatography System

Continuous bed chromatography is a new method of high performance liquid chromatography. It can be used for the separation and purification of solutes from low molecular mass substances to macromolecules such as proteins. The method has advantages in the ease of preparation, low backpressure at chromatographic runs, high resolution and large capacity for solutes.Although, there are a lot of automatic separation systems for bio-chromatography in the domestic and abroad, the vast majority is for the analytical separation or small-scale preparative separation, which is difficult to separate biological macromolecules. In2002, as one of new technology of the chromatographic separation, chromatographic separation of large-pore continuous-bed Cryogel appeared, which can achieve rapid and efficient separation of macromolecules, with the large porous media (Cryogel). Based on this, the preparative continuous bed chromatography for the super porous media was designed in this work.The design scheme of chromatography system for Cryogel is proposed on the basis of research on existing HPLC composition structure and working principle. It is a complex systematic project involving multiple disciplines of analytical chemistry, machinery, electronics, and automation. The system consists five parts, such as infusion module, auto-like module, separation module, detection module and the module of the control and recording.This work introduced the background of the subject in the first place, and then explored the basic principles of chromatography and development as well as domestic and international situation. On the basis of the understanding of the chromatography’s principle, it analyzed gel permeation chromatography instrument in the overall layout and details of the process, and designed the system control functions and accuracy requirements. This system included mechanical part and automatic control part. Mechanical part included the structural design of cabinet structure, the structural design of column chromatography, the structural design of the flow cell and the tructural design of multi-flow channel. Automatic control part included stepper motor driver circuit, the auto-sampler circuit, the data sampling circuit (pressure data acquisition circuit, temperature data acquisition circuit, the pH value acquisition circuit and the UV detector signal conditioning circuit), the serial communication interface circuit and the man-machine interface circuit design as the core of MCU AT89S52. According to the process and the function of the continuous bed chromatography, the system software included program design of automatically chromatography function artificial tomography features and calibration function (flow calibration, pressure calibration and pH value calibration), and designed the subroutine of the initialization, keyboard, display, digital filtering, multi-machine communication, achieving the harmonization of the hardware and software.The process control software of the host computer was developed based on the platform of Visual Basic. Users can operate directly and intuitively the system in the PC interface, which can communicate each control circuit communication of lower computer through the serial port, and make lower computer work according to the predetermined chromatographic separation process. The entire chromatography process is automatic and intelligent.Finally, complete machine assembly, experimental testing and performance assessment analysis to verify the functionality and accuracy of the system design further.