In this paper, we established a new on-line method using micelle to solvent stacking (MSS) technique combining with large amount sample electrokinetic stacking injection (LASEKSI) for the analysis of cationic molecules. In this MSS–LASEKSI, by modulating the integral EOF across the capillary, a equilibrium state was formed and can be maintained for a long time, leading to the continuous stacking of the analytes on the basis of MSS. Thereby, an extremely large amount sample was permitted to be injected into the capillary and then an improved enrichment fold can be achieved comparing with the each case. The variables affecting the performance of MSS–LASEKSI were investigated and discussed. Under the optimized conditions, 6.3×10³- and 6.4×10²-fold enrichment in peak heights upon normal CZE method (injected at 0.5psi for 3s) and number of plates of 2.9×10⁶ and 6.5×10⁵ were attained for berberine and theophylline, respectively. The developed method described here may provide prospects for exploiting a new concentration technique to achieve higher enrichment factor.