Abstract: OBJECTIVE To find out the characteristic peptide of Trachemys scripta elegans (Wied.) and establish a specific method for adulteration of tortoise-shell(Zhejiang tortoise shell). METHODS The samples were extracted by water extraction method, hydrolyzed by trypsin, searched for the characteristic peptide of Trachemys scripta elegans (Wied.) by ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) and data processing software, and sequenced by protein library searching technology. Ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-QQQ-MS) was used to establish the specific method based on the marker peptide of Trachemys scripta elegans (Wied.). RESULTS The marker peptidesm/z 400.23(n=2) of Trachemys scripta elegans (Wied.) was found. Using this characteristic ions, m/z 400.23(n=2)→374.05, 142.90, established an analysis method for identification of Trachemys scripta elegans (Wied.). In 20 samples, 11 samples detected Trachemys scripta elegans (Wied.) characteristic peptide. CONCLUSION This method has strong specificity and meets the technical requirements of analysis. This method can be used for the inspection of tortoise shell (Zhejiang tortoise shell) mixed with Trachemys scripta elegans (Wied.).