Study On Wide-bore Electrophoresis And Its Multi-dimensional Separation Systems

CE is one of the most efficient separation technologies, which has been extensively accepted and acknowledged in many fields. With the technology development, further study of the target has become a popular trend. However, the small sample loading capacity of the CE has turned to be its technical bottleneck in expanding its application. Consequently, CE has been restricted in analysis level so that it is difficult for CE in further process, such as transfer, collection, and even analysis through multi-dimensional high-resolution system.Firstly, for improvement the sample loading capacity, we adopted wide-bore tube with 1.0 mm id as separation channel, and introduced inner cooling mechanism into wide bore electrophoresis (WBE) to eliminate the Joule heat produced during the electrophoresis process. In WBE system, the inner cooling capillary was exactly fixed to the axial center of the wide-bore tube with the help of two hollow fiber membranes to strengthen the radiating efficiency. The result manisfested the method could improve the separation efficiency of the WBE. In addition, the sampling method based on WBE was explored and some samples were analyzed in WBE by hydrodynamic and electrokinetic injection. The results indicated the electrokinetic injection was feasible in WBE and the maximum sample loading amount was about 1.6μL by two-side hydrodynamic injection, which implied the WBE could be used as semi-preparing or preparing CE.Secondly, the larger sample loading capacity entitled the wide-bore electrophoresis system could be taken on sample transfer and further analysis operations. Taking advantage of the disparity between the inside diameter of wide-bore tube and the outside diameter of capillary, the two dimensional WBE-CE transfer system was built and zero dead volume connection was realized. Some model compounds certified it was feasible and controlled when the special three electrodes mode was adopted and field amplification was introduced. On the basic of the WBE-TC system, a multi-dimensional WBE-CE analysis platform integrated separation, transferring, collection, storage, offline and online enzyme reaction was built up. The alkaline phosphatase reaction model certified its feasibility. The results established the experimental foundation for the study of the wide-bore microchip. The experimental platform can facilitate the biological samples rapid separation and further analysis.Thirdly,another aim on establishing WBE system is to simplify two dimensional systems, especially HPLC-CE. WBE can directly separate and detect the effluent from HPLC, which avoided the sample deterioration during the collection, dried, or reconstitution processes in offline HPLC-CE. With the help of an injection valve, online two dimensional HPLC-WBE systems was built by coupling high performance liquid chromatography and the WBE system with large loadability and independent sampling system an and was called. Furthermore, the many sampling capillaries inWBE system design which can convert the space into time and shorten the interval in continue HPLC-WBE analysis. The current serial WBE analysis systems will broaden the application of electrophoresis technology, supply experimental basement for its development and made it possible for the analysis of complicated biological samples.