Interval isotachophoresis for purification and isolation of ionogenic species

A new preparative electrophoretic method in free solution is described, consisting of three consecutive steps: (i) filling the separation chamber with a suitable electrolyte system and sample in parallel streams by laminar hydrodynamic flow, (ii) applying the voltage across the chamber with isotachophoretic separation for a definite time interval operating in the direction perpendicular to that of filling, (iii) reapplying a hydrodynamic flow (without voltage) and collecting the separated species via an array of outlets. This approach completely eliminates the main drawback of the continuous flow electrophoresis (CFE), i.e., the electrohydrodynamic distortion of zones. This method utilizes the instrumentation devised for CFE and enables the isolation of large amounts of individual sample species comparable to that of CFE, with a resolution comparable to that of capillary isotachophoresis. The precise timing of the consecutive steps in the procedure as well as the stability of the operational parameters are of key importance for reproducibility. By using cationic isotachophoresis with 3 synthetic pI markers as model sample species, the reproducibility, stability and the separation power of the newly presented method are demonstrated. The sample throughput corresponds to micromoles per hour.