不同产地味连质量对比及近源植物分子鉴定研究

Study on Quality Comparison and Molecular Identification of Near Source Plants of Coptis chinensis from Different Producing Areas

目的应用ITS2条形码技术对味连、雅连等近源植物进行分子鉴定,对比分析不同产地味连药材质量差异性。方法提取味连样品总DNA,扩增ITS2序列并双向测序,计算种间距、种内Kimura-2-parameter(K2P)遗传距离,构建系统邻接(NJ)树。参照2020年版《中国药典》方法对味连药材进行检测。结果30批味连中,有7批含量不合格,8批总灰分不合格,25批镉、11批铜、3批铅超标,远大于其与雅连等近源植物的遗传距离。结论味连药材易出现有效成分含量和总灰分不合格,安全性指标方面易出现镉、铜、铅超标,药材质量方面需重点关注。ITS2序列分子鉴定能有效地鉴别味连药材到味连种,也为其它中药材的鉴定提供参考。

Objective To identificate Coptis chinensis,Coptis deltoidea and other near source plants by ITS2 bar code technology,compare and analyze the quality differences of Coptis chinensis from different producing areas.Methods The total DNA of the sample was extracted,the ITS2 sequence was amplified and sequenced in both directions.According to the Chinese Pharmacopoeia(2020 edition,Volume I and IV),Coptis chinensis was detected.Results Among the 30 batches collected,7 batches were unqualified for content,8 batches are unqualified for total ash,3 batches were unqualified for lead,11 batches were unqualified for Cadmium,25 batches were unqualified for copper.The maximum intra-genetic distances of Coptis chinensis was 0.003,it is far greater than the genetic distance between Coptis chinensis and its near source plants such as Coptis deltoidea.Conclusion The content and total ash content of Coptis chinensis are not up to standard,and cadmium,copper and lead are easy to exceed the standard in safety index.The ITS2 sequence and secondary structure can effectively identify the Coptis and its adulterans.It also provides reference for the identification of other Chinese medicinal materials and their near source plants.