We qauntitatively analyzed possible causes of the observed spreading of sample components which degrades separation compared with theoretical limits, and identified four potentially significant causes, electrokinetic dispersion, wall adsorption, enhanced diffusion due to Poiseuille flows driven by pressure gradients, and enhanced diffusion due to mobility variations associated with transverse temperature differences. Electrokinetic dispersion is caused by changes in the conductivity and pH distributions, proportional to the concentration of the sample relative to the buffer components, and independent of the tube diameter. One-dimensional numerical results for the separation of seven species in a sodium acetate buffer are presented to illustrate the effect. A detailed discussion of methods to control it is also presented. It is suggested that both wall adsorption of sample species and most coating methods used to control it or to reduce electroosmosis can be understood as aspects fo the Debye double-layer theory. Large molecules, with a high degree of ionization with the opposite sign to the wall charge, are preferentially attracted to the layer, excluding smaller molecules, and decreasing the wall potential and mobility. We demonstrate the importance of choosing a combination of pH and tube material such that the wall and the large proteins in the sample have the same charge sign and repel each other. The diffusion enhancement analysis is based on the analytic approximation of balancing the flow and mobility distrubance terms in the concentration equation for a sample species with the transverse diffusion term. This determines the fluctuating part of the concentration, with zero transverse average. The interaction of this concentration distribution with the fluctuations modifies the equation for the transverse average of the concentration, by adding diffusivity a 2 Y 2/48 D i· . Here a is the radius, D i is the diffusivity of the species, and Y is a disturbance speed and is a sum of contributions from the pressure-gradient-driven Poiseuille flow, the transverse mobility variation due to the heating in the tube, and other effects which we believe are smaller. The numerical factor of 48 is exact for the Poiseuille flow and the transverse mobility variations effects, and presumably it improves the estimate for the other effects. The Poiseuille flow profile is driven by variations in the electroosmotic slip velocity, which are mostly caused by conductivity variations along the tube and by changes in the Debye layer structure associated with composition changes along the tube. Contributions from temperature or radius variations along the tube are estimated, and are apparently smaller. Relatively insignificant causes of dispersion include sample dispersion by the difference between the electroosmotic flow and the slower flow in the Debye layer (the layer is to thin), variations in the tube radius, and convection and electrohydrodynamics flows. Molecular diffusion is important (and theoretical plate numbers are achieved) if other dispersion effects are small; it is larger for sample species with small molecules.