High-pressure carbon dioxide in contact with water dissolves to form carbonic acid, causing a decrease in pH. By use of these characteristics of a CO2/H2O biphasic system, chitosan-coated cationic liposomes of L-alpha-dipalmitoylphosphatidylcholine were successfully prepared by an improved supercritical reverse-phase evaporation (ISCRPE) method. Liposome-chitosan complexes carrying a positive charge were prepared in a single-step procedure without the use of acid or organic solvent, including ethanol. The maximum trapping efficiency of liposomes prepared by the ISCRPE method was 17%, with or without the addition of chitosan, compared to only 2% for liposomes prepared by the Bangham method. Furthermore, the liposomal dispersion was stable at room temperature in a sealed tube for over 30 days.