Calibration-free concentration determination of charged colloidal nanoparticles and determination of effective charges by capillary isotachophoresis.

Although colloidal nanoparticles show an electrophoretic heterogeneity under the conditions of capillary electrophoresis, which can be either due to the particle-size distribution and/or the particle shape distribution and/or the zeta-potential distribution, they can form correct isotachophoretic zones with sharp-moving boundaries. Therefore, the technique of isotachophoresis permits to generate plugs in which the co-ions and counter ions of the original colloidal solution are removed and replaced by the buffering counter ions of the leading electrolyte. It is shown that analytical isotachophoresis can be used to measure directly, without calibration, the molar (particle) concentration of dispersed ionic colloids provided that the transference number and the mean effective charge number of the particles (within the isotachophoretic zone) can be determined with adequate accuracy. The method can also be used to measure directly the effective charge number of biomacromolecules or colloidal particles, if solutions with known molar (particle) concentration can be prepared. The validity of the approach was confirmed for a model solution containing a known molar concentration of bovine serum albumin.