Abstract: In order to explore the functional components of donkey bone collagen and the repair effect of donkey bone collagen on the damaged skin barrier. The functional composition of donkey bone collagen was analyzed by UPLC-Orbitrap MS technique, the effects of donkey bone collagen on the proliferation of HaCaT cells were detected by Brdu method, and the effects on the migration of HaCaT cells were detected by scratch method. The results showed that the protein content of donkey bone collagen was 57.5%, which had a typical collagen amino acid composition. The donkey bone collagen contained abundant functional ingredients such as amino acids, vitamins, sugar and lipids. Donkey bone collagen could significantly promote the proliferation and migration of HaCaT cells, and the effect of 0.15 g/mL donkey bone collagen in promoting the proliferation and migration of HaCaT cells was similar to that of 5% fetal bovine serum, which was significantly better than the blank control group (P<0.05). It is inferred that donkey bone collagen could accelerate the repair of skin barrier, and its mechanism might be related to promoting keratinocytes proliferation and migration.