Duan Ya-Ping, Luo Jiao-Yang, Dou Xiao-Wen, He Liu, Li Kun-Lun, Yang Shi-Hai, Yang Mei-Hua
College of Traditional Chinese Medicine, Jilin Agricultural University, Changchun 130118， China.
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100193, China.
Zhongguo Zhong Yao Za Zhi. 2018 Aug;43(15):3105-3114. doi: 10.19540/j.cnki.cjcmm.20180510.005.
With the extensive clinical application of Tripterygium wilfordii, there are many counterfeit products on the market. Traditional technology can not effectively identify the authenticity of the traditional Chinese medicine. Therefore, a strategy of accurate identification and quality evaluation of Tripterygium based on DNA barcode and chemical fingerprint spectrum was established. Based on DNA barcode technology, HMMer annotation method of hidden Markov model and K2P model were used to analyze genetic distance.BLAST1, nearest distance and phylogenetic tree (NJ-tree) methods were used to assess the identification efficiency of the ITS2 barcode. The fingerprint of 27 T. wilfordii was established by UPLC-PDA method, and the similarity of the fingerprint of different sources was evaluated. The main components of T. wilfordii were determined by LC-MS/MS. The results revealed that the intraspecific genetic distances of T. wilfordii were lower than the interspecific genetic distances between T. wilfordii and its adulterants. The results of similarity search showed that ITS2 sequence was used to identify T. wilfordii and its adulterants. The clustering of T. wilfordii and its adulterants was clear in the tree of NJ cluster, and 12 of 27 samples were identified as true T. wilfordii.The chemical fingerprint spectrum research indicates that the feature one region can distinguish the false product of tripterygium glycosides more intuitively. The cluster analysis of HCA-thermal map showed that the contents of six active components of T. wilfordii from different habitats were significantly different, which could be used to evaluate the quality of T. wilfordii. This paper is of guiding significance for the accurate identification and quality evaluation of Tripterygium medicinal plants.
随着雷公藤临床应用的广泛开展，市场上出现了许多假冒伪劣产品。传统技术无法有效鉴别中药的真伪。因此，建立了基于DNA条形码和化学指纹图谱的雷公藤精准鉴定与质量评价策略。基于DNA条形码技术，采用隐马尔可夫模型的HMMer注释方法和K2P模型分析遗传距离。运用BLAST1、最近距离和系统发育树（NJ树）方法评估ITS2条形码的鉴定效率。采用超高效液相色谱-光电二极管阵列检测法（UPLC-PDA）建立了27批雷公藤的指纹图谱，并对不同来源指纹图谱的相似度进行了评价。通过液相色谱-串联质谱法（LC-MS/MS）测定了雷公藤的主要成分。结果表明，雷公藤种内遗传距离低于其与伪品之间的种间遗传距离。相似度搜索结果表明，ITS2序列可用于鉴别雷公藤及其伪品。在NJ聚类树中，雷公藤及其伪品的聚类清晰，27个样品中有12个被鉴定为正品雷公藤。化学指纹图谱研究表明，特征一区能更直观地区分雷公藤多苷的伪品。热图聚类分析表明，不同产地雷公藤6种活性成分的含量存在显著差异，可用于评价雷公藤的质量。本文对雷公藤药用植物的精准鉴定和质量评价具有指导意义。
