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Lipid-like pseudo-double-chained catanionic surfactants have emerged as the attractive materials to prepare potential vesicular carriers in drug and gene delivery applications. In particular, the semispontaneous process has been developed to fabricate ethosome-like catanionic vesicles for the transdermal drug delivery. In this work, Arbutin (a water-soluble drug) encapsulation efficiency of ethosome-like catanionic vesicles fabricated from decyltrimethylammonium-tetradecylsulfate (DeTMA-TS, CH3(CH2)9 N(CH3)3-CH3(CH2)13SO4) and decyltrimethylammonium-dodecylsulfate (DeTMA-DS, CH3(CH2)9N(CH3)3-CH3(CH2)11SO4) with various amounts of ethanol and cholesterol in tris buffer solution was experimentally determined. A simple unilamellar vesicle (ULV) model, resulting in the theoretical encapsulation efficiency within ±10% error for most vesicle compositions, was also developed. Such agreement indirectly confirmed the formation of unilamellar vesicles by the preparation method. Stable ethosome-like catanionic vesicles by using catanionic surfactants with the aid of suitable amounts of ethanol and cholesterol, which led to polydispersity index (PDI) values of vesicle size distribution less than 0.3, were successfully prepared and their hydrophilic drug encapsulation efficiencies can be accurately predicted. Furthermore, the linear correlations of the trap volume ratio with both vesicle size and concentration of the extra added CHOL also provide important guidelines for controlling the drug loading of ethosome-like catanionic vesicles. The accomplishments reached for the novel vesicles are useful for developing their transdermal drug delivery applications. [ABSTRACT FROM AUTHOR]