The aim of this study was to establish an authenticity identification method for colla corii asini and quantitatively analyze donkey-derived components in colla corii asini samples. The DNA purification column was used to establish the extraction method of genomic DNA from colla corii asini. And the standards of colla corii asini and common fakes were used to verify the specificity of primers. The target fragment was cloned, and the recombinant plasmid pGMT-Equ66bp was used to construct a positive control standard. The detection limit of donkey-derived DNA components of colla corii asini and standard curve was tested, and 22 batches of colla corii asini were analyzed. The genomic DNA concentration of colla corii asini was (82.17±0.09)-(172.81±1.81) ng/μL, and A260/A280 was (1.561±0.07)-(1.821±0.04). The results of specific experiments showed that the designed primers had strong specificity with CT value less than 32. For the 22 samples, CT values was between 19.83 and 28.85, which indicating that all above samples were genuine colla corii asini, which was consistent with the results of mass spectrometry in Chinese Pharmacopoeia. In this study, molecular biology technology was used to effectively extract the genomic DNA components of colla corii asini, and a quantitative analysis system of donkey-derived DNA components in colla corii asini was successfully constructed. It could effectively identify the authenticity of colla corii asini and was helpful for the implementation of quality control of colla corii asini.
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