市售中药饮片DNA条形码鉴定研究  

DNA barcoding identification of commercial decoctions in traditional Chinese medicine

作　　者：辛天怡 娄千 郝利军 李冉郡 林余霖 许文杰 马婷玉 林烨暄 董一昕 蒋澄宇[3] 宋经元 XIN Tian-yi;LOU Qian;HAO Li-jun;LI Ran-jun;LIN Yu-lin;XU Wen-jie;MA Ting-yu;LIN Ye-xuan;DONG Yi-xin;JIANG Cheng-yu;SONG Jing-yuan(Key Lab of Chinese Medicine Resources Conservation,State Administration of Traditional Chinese Medicine of the People's Republic of China,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100193,China;School of Life and Science,Southwest Jiaotong University,Chengdu 610031,China;State Key Laboratory of Medical Molecular Biology,Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences,Department of Biochemistry,Peking Union Medical College,Beijing 100005,China;Engineering Research Center of Chinese Medicine Resource,Ministry of Education,Beijing 100193,China)

机构地区：[1]中国医学科学院、北京协和医学院药用植物研究所,国家中医药管理局中药资源保护重点研究室,北京100193 [2]西南交通大学生命科学与工程学院,四川成都610031 [3]中国医学科学院基础医学研究所医学分子生物学国家重点实验室,北京协和医学院基础学院生物化学系,北京100005 [4]中药资源教育部工程研究中心,北京100193

出　　处：《药学学报》2021年第3期879-889,共11页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金青年基金项目(82003906);国家重点研发计划(2019YFC1711100)。

摘　　要：中药材DNA条形码分子鉴定法指导原则已纳入《中国药典》,但目前尚缺乏应用于中药饮片的系统性研究。本研究共收集212份市售中药饮片,包含根及根茎、果实种子、全草、花、叶、皮、茎木等不同入药部位,验证中药材DNA条形码分子鉴定法对中药饮片基原物种鉴定的适用性和准确性。结果表明,共有161份样本成功获得DNA条形码序列,占收集中药饮片样本量的75.9%,其余样本基因组DNA降解,无法获得扩增产物。实验所得DNA条形码序列分析结果如下:138份饮片为《中国药典》(2020年版)收载相应药材的基原物种,占获得序列样本数的85.7%;14份样本分析结果中包含《中国药典》收载相应药材的基原物种以及同属近缘物种,经过形态学鉴定,其中8份为正品,3份掺有伪品,3份缺少鉴别特征,不能确定基原;7份样本为混伪品;另有2份样本检出掺伪现象。本研究证实中药材DNA条形码分子鉴定法能有效检出市售中药饮片的掺伪/混伪现象,可用于市售中药饮片的基原物种鉴定,值得在中药饮片加工生产企业及药品监督检验管理机构推广,同时有助于促进中药学研究的跨学科交流,共同挖掘中药作用靶点和药效机制,推动个性化精准医疗及"精准药物研发"。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.

关 键 词：中药饮片 DNA条形码 ITS2 PSBA-TRNH 分子鉴定 市场监控 

分 类 号：R931[医药卫生—生药学]