This study was conducted to evaluate physiological activity of Robinia pseudo-acacia leaf and its skin penetration using liposome and cell penetrating peptide. After extraction with Robinia pseudo-acacia leaf using the distilled water and supercritical, various physiological activities were examined. In antioxidants experiments, the total concentration of polyphenol compounds was determined to be 56.88 mg/g in hydrothermal extract, 45.07 mg/g in supercritical extract. In the antimicrobial experiments, the hydrothermal extract had no effect, but supercritical extract represented maximum clear zone of 14.00 mm in Staphylococcus aureus strain. Liposome containing the RSE (Robinia pseudo-acacia leaf supercritical extract) reduced particle size and stabilized zeta potential. In the epidermal permeability experiment, it was confirmed that the permeation of liposome containing the RSE and cell penetrating peptides was remarkable.