In order to improve the stability and transdermal efficacy of ethosomes, binary ceramide modified ethosomes was constructed by the combination of propylene glycol and ethanol. Furthermore, lipid soluble active substance, retinol palmitate (RP) and water-soluble active substance, carnosine (Car) were further selected to prepare RP-Car binary ceramide modified ethosomes (RP-Car binary cerethosomes), and the storage stability and transdermal properties were investigated. The results showed that when the volume ratio of ethanol to propylene glycol was 6:4, the encapsulation efficiency of RP in the RP-Car binary cerethosomes was (80.28 ± 0.10)%, and the encapsulation efficiency of Car was (47.20 ± 0.71)%; The TEM results showed that the RP-Car binary cerethosomes were relatively regular spherical vesicles with a particle size of about 70 nm; The RP-Car binary cerethosomes showed no significant changes after 120 days of storage at 4 ℃, 25 ℃ and 45 ℃; The results of in vitro transdermal experiment and laser confocal laser showed that the skin accumulation and permeability of RP and Car increased, and the skin accumulation was (0.273 ± 0.002) μg/cm2 and (22.521 ± 0.280) μg/cm2, respectively, the skin permeability was 0 μg/cm2 and (20.559 ± 0.120) μg/cm2, respectively.