Identification of vinegar-quenched Trionycis carapax in Gejie Dingchuan Pills by real-time PCR
AIM To establish a DNA molecular identification method for vinegar-quenched Trionycis Carapax in Gejie Dingchuan Pills.METHODS Two specific primers(TS and AF)were designed according to the difference of mitochondrial gene sequence between Trionyx sinensis and Apalone ferox,and real-time PCR amplification and method ological investigation were carried out.The established method was used to identify the T.Carapax in commercially available Gejie Dingchuan Pills.RESULTS The fluorescence signals of DNA samples containing T.sinensis and A.ferox could only be detected after amplification with the corresponding primers,and there was a specific temperature peak(Tm was 77.45℃or 79.72℃).The assay exhibited a good specificity and high repeatability(CV<1%),and limit of detection was above 10 copies/μL.Among the 10 batches of Gejie Dingchuan Pills,the presence of T.sinensis but not A.ferox were detected in nine of them,which were identified as authentic.Another batch was found to contain both T.sinensis and A.ferox,which was identified as adulterant.CONCLUSION The established method in this study can quickly and reliably identify vinegar-quenched T.Carapax in Gejie Dingchuan Pills,which is helpful to monitor the quality of Chinese Patent Medicines(CPM)containing vinegar-quenched Trionycis Carapax,and provides a new approach for specific identification of animal-derived components in CPM.
国家科技期刊平台
NSTL
万方数据
维普
