To establish HPLC (High Performance Liquid Chromatography) fingerprint of Carapax et Plastrum Testudinis by identifying tortoiseshells and their adulterants. Methods:A total of 21 batches of tortoiseshells and their adulterants from different habitats was collected，and 42 samples were classified by the carapace and plastron. The pre-treatment of acid hydrolysis，dephosphorization and decalcification pretreatment with 6-aminoquinolyl-N-hydroxysuccinimidyl-carbamate (AQC) derivatization reaction was carried out using the C18 column (3.9×150 mm，3 μm)，the column temperature of 37 ℃. Acetonitrile，water，salt buffer were used as mobile phase for gradient elution at a flow rate of 1.00 mL/min and a the detection wavelength of 248 nm. The system of similarity evaluation (Pharmacopoeia 2012) was used to analyze tortoiseshells by the carapace and plastron，tortoisshells and their common adulterants. Results:The fingerprint had good precision，reproducibility and stability. The similarity of tortoiseshells was between 0.932-0.995. The percentage was less than 90.91% under the similarity of 0.90 between tortoiseshells of their adulterants. Conclusion:The difference of HPLC is relatively small between tortoiseshells' carapaces and plastrons，and the difference between tortoise shells and their adulterants is obvious. The established HPLC fingerprint can be used for the quality evaluation of tortoiseshells.