Continuous processes, electrophoresis

CE is generally more suited to analytical separations than to preparative-scale separations. However, given the success of CE methods for chiral separations, it seems reasonable to explore the utility of preparative electrophoretic methods to chiral separations. Thus, the purpose of this work is to highlight some of the developments in the application of preparative electrophoresis to chiral separations. Both batch and continuous processes will be examined. 

Ultimately, however, it should be noted that these examples of classical gel electrophoretic separations are batch processes and therefore limited in sample throughput. To achieve true preparative-scale separations by electrophoresis, it becomes necessary to convert to continuous processes. 

Figure 7.3. In continuous deflection electrophoresis, flow down a paper sheet or between closely spaced plates carries different solutes, which are gradually separated by electrophoresis along axis x, to different collection ports. However, the flow is classified as passive because it does not enhance the separation occurring along the electrical-field axis. The role of flow is to aid continuous sample collection, not to fundamentally alter the separation process.

Many 2D planar structures have been used to implement deflection (continuous flow) electrophoresis. The primary requirement is that flow and electrophoresis be carried out simultaneously and uniformly. Hanging paper curtains soaked with electrolyte and fed a stream of electrolyte from above served admirably for this purpose when the technique was initiated in the 1950s. In recent years thin flow channels enclosed between flat plates have become important. The process is complicated by parabolic flow, which distorts and effectively broadens the electrophoretic zones. More detail is available in the cited references on electrophoresis [3-5]. 

The simulated moving bed adsorption technique is based on the movement of the stationary phase. The front and back ends of a series of columns are connected to form a circle, and during rotation of the columns a countercurrent movement of the phase relative to the liquid stream in the system is developed [158]. Injections of the fresh chiral analyte and the solvent are made at various coimecting points, and the separated enantiomers are withdrawn simultaneously at certain time intervals. This is a continuous process that provides certain advantages for enantiodiscrimination. The chiral selectors used in this technique are the same as those utilized in liquid chromatography and capillary electrophoresis. 

Continuous free flow electrophoresis has been used for the separation of biopolymers (e.g. ovalbumin and lysozyme) [20] as well as smaller inorganic species (e.g. [Co sepulchrate)] and [Co (CN)g] ) [21]. Sample processing rates of 15 mg h were reported for a mixture of Amaranth (MW 804) and Patent Blue VF (MW 1159) [22]. 

Four different electrokinetic processes are known. Two of them, electroosmosis and electrophoresis, were described in 1809 by Ferdinand Friedrich Renss, a professor at the University of Moscow. The schematic of a cell appropriate for realizing and studying electroosmosis is shown in Fig. 31.1a. An electrolyte solution in a U-shaped cell is divided in two parts by a porous diaphragm. Auxiliary electrodes are placed in each of the half-cells to set up an electric held in the solution. Under the inhuence of this held, the solution starts to how through the diaphragm in the direction of one of the electrodes. The how continues until a hydrostahc pressure differential (height of liquid column) has been built up between the two cell parts which is such as to compensate the electroosmotic force. 

Kasicka, V., Pruslk, Z., Sazelova, P., Jiracek, J. and Barth, T., Theory of the correlation between capillary and free-flow zone electrophoresis and its use for the conversion of analytical capillary separations to continuous free-flow preparative processes. Application to analysis and preparation of fragments of insulin, ]. Chromatogr. A, 796, 211, 1998. 

On the other hand, the most severe constraint of CL analyses is their relatively low selectivity. One major goal of CL methodologies is thus to improve selectivity, which can be accomplished in three main ways (1) by coupling the CL reaction to a previous, highly selective biochemical process such as an immunochemical and/or enzymatic reaction (2) by using a prior continuous separation technique such as liquid chromatography or capillary electrophoresis or (3) by mathematical discrimination of the combined CL signals. This last approach is discussed in Sec. 4. 

Some coupled systems allow measurement of the main N and P forms (nitrate, ammonia and orthophosphates) [22,27,29], among which is a system based on membrane technology in combination with semi-micro continuous-flow analysis (pCFA) with classical colorimetry. With the same principle (classical colorimetry), another system [30] proposes the measurement of phosphate, iron and sulphate by flow-injection analysis (FIA). These systems are derived from laboratory procedures, as in a recent work [31] where capillary electrophoresis (CE) was used for the separation of inorganic and organic ions from waters in a pulp and paper process. Chloride, thiosulphate, sulphate, oxalate,... 

Electrophoresis can also be conducted on-line, as an element of industrial process monitoring and/or control. In this case a slip-stream sample is usually withdrawn from a process vessel, diluted in a mixing tank to reduce the sample turbidity, and then pumped through an electrophoresis cell that is fitted with stop-flow solenoid valves. The flow is stopped for long enough to make an electrophoresis measurement and then resumed. The sampling can be either intermittent or continuous. An example is described in reference [265]). 

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