Capillary Isotachophoresis (CITP) is an advanced technique that is used for the separation of substances with electrophoretic mobility and their charges. The isotachophoresis electrolyte system is presented, along with how the effective mobilities and molar amounts of the analytes influence the separation and time required to attain the steady state for a certain electrolyte condition. One approach called CITP uses a “moving border.” Between the leading and terminating electrolytes is a sample zone where all solutes travel at the same speed in distinct bands. In this article, we have described the different and latest developments of CITP, i.e., isotachophoresis in nanoparticle studies, in the hyphenation technique, in the bioanalytical sector, and in various food tracing elements. Although traditional Isotachophoresis is becoming less popular, isotachophoresis is becoming more used for a variety of samples as a preconcentration approach for CZE analysis.