Analysis Of Ions And Proteins By Wide-bore Electrophoresis

Capillary electrophoresis (CE) has become one of the most advanced separation techniques in recent years due to its high resolution, online detection, fast analysis speed , a variety of separation modes and easy to couple to mass spectrometer, it has found widespread application in analytical, biological chemistry, biotechnology , pharmaceutical and biomedical analysis.However,because of the Joule heat, just the small ID(50μm or 75μm)of the capillary was used in electrophoresis, which limits the sensitivity and sample loading so that the lower components can't be found or separated, the tiny sample loading capacity becomes the major obstacle to extending the application scope of capillary electrophoresis.In order to increase the sample loading capacity and improve the detection sensitivity, in our work, a new device was set up. In the device, a quartz tube of 1000μm inner diameter was used as separation channel, and a capillary of 690μm outer diameter was inserted to the center of the quartz tube as inner cooling capillary, the Joule heat generated during electrophoresis was removed timely when coolant passed through the cooling capillary.In this paper, the capillary zone electrophoresis(CZE)has been selected to evaluate the Wide-bore system, when the experimental conditions were optimized (such as the pH and concentration of buffer, the flow rate and temperature of cooling water, additive concentration , separation voltage), six kinds of organic acids, three kinds of organic base and the proteins were separated successful,and the reprodu- cibility of results is satisfactory.Capillary isoelectric focusing (CIEF) is a powerful separation technique for the analysis of proteins. In this paper, we transferred the mode of CIEF to the Wide-bore system . In order to obtain the best separation results, several kinds of conditions were optimized, such as the focusing power, the flow rate of inner cooling water, the optimum detection wavelength, the appropriate buffers , the additive concentration and concentration of carrier ampholyte. Under optimized conditions, conalbumin, carbonic anhydrase and ribonuclease could be detected well.