Abstract: To study the source of different gelatins of donkey-hide gelatin lumps, four kinds of donkey-hide gelatin lumps were added with low content (1 mg/mL) of bovine gelatin and porcine gelatin respectively. High-performance liquid chromatography-mass spectrometry (HPLC-MS/MS) were chosen to identify the characteristic peptides of donkey-hide gelatin, bovine gelatin and porcine gelatin. The results showed that 18, 3 and 5 common characteristic peptides were accurately detected in the four kinds of donkey-hide gelatin lump for donkey-hide gelatin, bovine gelatin and porcine gelatin, respectively, with 10, 1 and 1 from the α₁ chain and 8, 2 and 4 from the α₂ chain. Meanwhile, the number of common characteristic peptides with hydroxylated modifications was 15, 2 and 4 respectively, while those without any modification groups were 3, 1 and 1. These common characteristic peptides would provide a basis for the HPLC-MS/MS traceable identification of the gelatin source in donkey-hide gelatin lump, which is of reference significance for the quality assurance of donkey-hide gelatin lump products.