RESUMEN
Although the guiding principles for molecular identification of traditional Chinese medicines (TCM) using DNA barcoding have been recorded in the Chinese Pharmacopoeia, there is still a lack of systematic research on its application to commercial TCM decoctions. In this study, a total of 212 commercial TCM decoctions derived from different medicinal parts such as root and rhizome, fruit and seed, herb, flower, leaf, cortex, and caulis were collected to verify applicability and accuracy of the method. DNA barcodes were successfully obtained from 75.9% (161/212) of the samples, while other samples failed to be amplified due to genomic DNA degradation. Among the 161 samples, 85.7% of them were identified as recorded species in the Chinese Pharmacopoeia (2020 edition). In addition, 14 samples could be identified as species recorded in the Chinese Pharmacopoeia and their closely related species in the same genus. Morphological identification for the unconfirmed samples showed that eight were genuine species and three were adulterants, while the other three were unidentifiable due to lack of morphological characteristics. Furthermore, the DNA barcodes of seven samples accurately mapped to the sequences of adulterants. Remarkably, counterfeit products were detected in two samples. These results demonstrate that DNA barcoding is suitable for the identification of commercial TCM decoctions. The method can effectively detect adulterants and is appropriate for use throughout the industrial chain of TCM production and distribution, and by the supervisory agencies as well.
RESUMEN
Adulterants and counterfeits were found in some of the commercial traditional Chinese medicine (TCM) decoctions in Hongjin Xiaojie Jiaonang, Hongjin Xiaojie Pian, and Chaihuang Keli during the national drug sampling inspection. However, it was difficult to determine the species of the adulterants and counterfeits by conventional testing methods. Therefore, a total of 184 samples of the TCM decoctions and raw materials belong to the prescriptions of above mentioned traditional Chinese patent medicines, including Bupleuri Radix, Bajiaolian, Heimayi, and Shufuchong, were collected and authenticated by DNA barcoding technology. 111 ITS2 sequences were obtained from 115 commercial TCM decoctions and raw materials of Bupleuri Radix, among which 71 were Bupleurum chinense, three were B. scorzonerifolium, and 31 were closely related species in the same genus. In addition, counterfeits derived from different genera, such as Ailanthus altissima (one sample), Saposhnikovia divaricate (two samples), and Solidago decurrens (three samples), were also detected. 21 ITS2 sequences were obtained from 22 commercial TCM raw materials of Bajiaolian, among which 15 were Diphylleia sinensis and six were Dysosma versipellis and other species in genus Dysosma. For 22 Heimayi samples, PCR amplification of COI sequence was failed due to genomic DNA degradation. Among 38 Shufuchong samples, 24 COI sequences were obtained and only nine of them were the genuine species (Armadillidium vulgare) recorded in the Chinese Pharmacopoeia, 11 were Porcellio laevis, two were Mongoloniscus sinensis, and two samples could not be identified due to the limitation of database. This study demonstrates that DNA barcoding technology is suitable for the species authentication of the decoctions of traditional Chinese patent medicine prescription. It is a conductive way for the establishment of traceability system for the whole TCM industrial chain.
RESUMEN
OBJECTIVE: To identify the commercial medicinal materials and decoction pieces of Lycii Cortex and its adulterants using DNA barcoding technology. METHODS: A total of 137 samples, including 105 voucher samples belonging to nine species, seven GenBank sequences, and 25 test samples were involved in this study. Experiments were performed in accordance with the DNA barcoding standard operating procedures (DNA barcoding SOP) to get the ITS2 sequences. A DNA barcode database of Lycii Cortex and its adulterants were successfully constructed using 112 ITS2 sequences, which were amplified from the voucher samples and downloaded from the GenBank. This database was used to identify the commercial medicinal materials and decoction pieces of Lycii Cortex. RESULTS: The lengths of the ITS2 regions of the Lycii Cortex were 212-230 bp. The ITS2 sequences could clearly distinguish Lycii Cortex and its adulterants. Fifty percent of the commercial samples gained the ideal genomic DNA for the sequence amplification. Using the established database, the above-mentioned sequences were authenticated as Lycium chinense. CONCLUSION: ITS2 Sequence may be a suitable marker for the identification of Lycii Cortex and its adulterants. The DNA barcode databaseof Lycii Cortex and its adulterants constructed in this study are able to successfully identify the raw materials of the commercial medicinal materials and decoction pieces of Lycii Cortex that are currently available in the market.