Capillary flow band broadening

The electroosmotic flow profile is very different from that for a phase moving under forced pressure. Figure 12.40 compares the flow profile for electroosmosis with that for hydrodynamic pressure. The uniform, flat profile for electroosmosis helps to minimize band broadening in capillary electrophoresis, thus improving separation efficiency. 

The flow profiles of electrodriven and pressure driven separations are illustrated in Figure 9.2. Electroosmotic flow, since it originates near the capillary walls, is characterized by a flat flow profile. A laminar profile is observed in pressure-driven systems. In pressure-driven flow systems, the highest velocities are reached in the center of the flow channels, while the lowest velocities are attained near the column walls. Since a zone of analyte-distributing events across the flow conduit has different velocities across a laminar profile, band broadening results as the analyte zone is transferred through the conduit. The flat electroosmotic flow profile created in electrodriven separations is a principal advantage of capillary electrophoretic techniques and results in extremely efficient separations. 

The choice of carrier gas and gas flow control are critical for successful GC. The carrier gas does no more in the separation process than its name implies it carries the vapor phase analyte molecules along the column. As such, it must be inert, non-toxic, inexpensive, highly pure and must provide efficient transport with minimal band broadening. For packed column GC, nitrogen is the most commonly used carrier gas, followed by helium. For capillary column GC, the most common carrier gas is helium, followed by hydrogen and nitrogen. 

The electrolyte composition and pH should not be too detrimental. Aspects such as pressure, temperature, and injection technique may have an influence. Concerning the interface, the sheath flow should be optimized with respect to composition, pH, and flow rate. Furthermore, the positions of the fused-silica capillary as well as the API probe need to be carefully optimized. The mixing volume at the tip of the capillary must be kept at a minimum to avoid postcolumn band broadening caused by diffusion. 

The most common HPLC column diameter is 4.6 mm. There is a trend toward narrower columns (2 mm, 1 mm, and capillary columns down to 25 pm) for several reasons. Narrow columns are more compatible with mass spectrometers, which require low solvent flow. Narrow columns require less sample and produce less waste. Heat generated by friction of solvent flow inside the column is more easily dissipated from a narrow column to maintain isothermal conditions. Instruments must be specially designed to accommodate column diameters <2 mm or else band broadening outside the column becomes significant. 

The EOF and make-up flow were combined at the exit of the capillary before nebulisation. In this way, no suction was observed and band broadening was not significant. Alkali, alkaline-earth and heavy metal ions were analyzed at concentrations of 2-100... 

In terms of organization, the text has two main parts. The first six chapters constitute generic background material applicable to a wide range of separation methods. This part includes the theoretical foundations of separations, which are rooted in transport, flow, and equilibrium phenomena. It incorporates concepts that are broadly relevant to separations diffusion, capillary and packed bed flow, viscous phenomena, Gaussian zone formation, random walk processes, criteria of band broadening and resolution, steady-state zones, the statistics of overlapping peaks, two-dimensional separations, and so on. 

This technique has proven to be a powerful separation technique for the separation of macromolecules, such as antibodies, for two reasons the near flat plug flow profile and the small diffusion constant of the antibodies. These characteristics eliminate band broadening. With both superior separation power and high detection sensitivity, capillary electrophoresis (CE) can separate free Ab and Ag from bound Ab and Ag rapidly and is suitable for immunoassays. CE can combine immunologic recognition with on-line quantitation, microscale... 

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