The invention discloses donkey characteristic polypeptide and application thereof to detection on donkey skin derived ingredients. The amino acid sequence of the donkey characteristic polypeptide provided by the invention is shown as SEQ ID NO.1 and the polypeptide is specifically contained in collagen of a donkey. Moreover, the invention also provides a method for detecting donkey skin derived ingredients in glue traditional Chinese medicine, which comprises: carrying out enzyme digestion on the glue traditional Chinese medicine by adopting trypsin or arginine-C enzyme so as to enable the characteristic polypeptide to be dissociated from the collagen; then carrying out detection by adopting liquid chromatography-tandem mass spectrometry so as to judge whether the donkey skin derived ingredients are included in the glue traditional Chinese medicine. The method has the advantages of strong characteristic performance, high sensitivity, simplicity for operation and the like and can accurately authenticate whether the donkey skin derived ingredients are included in the glue traditional Chinese medicine.