Type I collagen peptides were prepared by hydrolysis of type I collagen protein with different enzyme combinations and this study found that the peptides obtained by papain/pepsin/trypsin combination hydrolysis were of the strongest anti-X-ray capability. The collagen peptides-carboxymethyl chitosan (CP-CMC-CaCl2) microspheres were prepared by emulsification method and used to investigate the effects on mice skin damages in vitro and in vivo. In vitro experiments indicated that the CP-CMC-CaCl2 microspheres had a protective effect on the X-ray irradiated damages of mice thymus lymphocytes. The mice skin surface texture morphological and histomorphological analysis showed a consistent result that the CP-CMC-CaCl2 microspheres played a certain role in the prevention of UV-B irradiated skin damages and could also improve ultraviolet prevention ability. Besides., the preventive function was better than its repair function. It is possible to improve the preventive function to skin photoaging and play a better effectiveness of cosmetic skin care by adding the CP-CMC-CaCl2 microspheres into make-up cosmetics as a natural antioxidant resource.