A novel catanionic surfactants vesicle system composed of octyltriethylammonium bromide/ sodium dodecyl benzene sulfonate (C8NE3Br/SDBS) has been developed as pseudostationary phase (PSP) in EKC. The C8NE3Br/SDBS system possesses a large vesicle phase region and none of agglomeration phenomena appeared while mixing cationic and anionic surfactants at any molar ratio. Electrophoretic and chromatographic parameters including elution window, hydrophobic selectivity, polar group selectivity, and shape selectivity were characterized using the vesicle at molar ratio of C8NE3Br to SDBS of 3: 7 as PSP. Compared with SDS micelles, the vesicle PSP possessed a wider elution window and a better selectivity. The retention behavior and selectivity differences between the novel vesicle and SDS micelles were evaluated through linear solvation energy relationship (LSER) analysis. Though the cohesiveness and the hydrogen bond acidity have greatest influences on the solutes retention and selectivity in both the vesicle and SDS micelle, the vesicle PSP demonstrated a higher hydrophobicity and a lower hydrogen bonding donating capability owing to compact bilayer structure of vesicle. Additionally, the vesicle system had a stronger hydrogen bond accepting capability than SDS micelle. Consequently, according to LSER analysis, the bigger coefficients for v, b, and a revealed the vesicle PSP had a better separation selectivity than conventional SDS micelle.