Abstract: To explore the pharmacological activity of donkey hide gelatin related protein components, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Orbitrap Fusion Lumos Tribrid were used to reveal the composition of Colla Corii Asini (CCA), which would provide a basis for the exploration of protein components related to the pharmacological activity of CCA and further development of CCA. The results showed that the molecular weight was distributed in the range of 10~250 kDa. By comparing with the equine protein library, 316 proteins were retrieved. Results found that CCA contained a variety of active proteins, including leucine-rich proteoglycans, such as decorin, biglycan, Lumican, fibromodulin, and other active substances such as collagen and immunoglobulins. Further bioinformatics analysis of proteins in CCA revealed that these proteins are participate in signal pathways such as systemic lupus erythematosus, protein digestion and absorption, ECM receptor interaction, viral carcinogenesis, hypertrophic cardiomyopathy, and dilated cardiomyopathy. The screening of pharmacologically active ingredients provides a theoretical basis. This results would provide a theoretical basis for the screening and further development of pharmacological active components of CCA.