基于COI序列的水牛角及其易混伪品DNA条形码鉴定研究  

Identification of water buffalo horn and its adulterants using COI barcode

作　　者：刘旭朝[1,2] 周丽思 刘金欣[1,3] 贾静 宋经元 石林春 

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,中草药物质基础与资源利用教育部重点实验室,北京100193 [2]山东中医药大学药学院,山东济南250355 [3]承德医学院,河北省中药研究与开发重点实验室,河北承德067000

出　　处：《药学学报》2017年第3期494-499,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81303158);国家“重大新药创制”科技重大专项资助项目(2014ZX09304307001)

摘　　要：水牛角作为犀角的代用品是急症用药安宫牛黄丸的主要成份,近年来常有混伪品流入市场,急需建立有效鉴定方法。本研究共收集155份原动物及市售水牛角样品,通过优化DNA提取方法,PCR扩增、双向测序、序列拼接获得153条COI序列。93份原动物COI序列经条形码间隔法和建树法核验后纳入中药材DNA条形码动物药材数据库,利用中药材DNA条形码鉴定系统(www.tcmbarcode.cn)对62份市售水牛角药材进行鉴定。除2份市售药材无法获得COI序列外,54.8%的市售药材为水牛角,29%的市售药材为牦牛角。本研究表明牦牛角为市售水牛角药材主要伪品来源,DNA条形码技术可用于区分水牛角及其易混伪品,应加强市场监管,以确保急症用药临床疗效。Bubali cornu（water buffalo horn） has been used as the substitute for Cornu rhinoceri asiatici（rhino horn） in clinical applications, and is the essential ingredient of Angong Niuhuang Wan. In recent years, there are a number of adulterants on the commercial herbal medicine markets. An efficient tool is required for species identification. In this study, 155 Bubali cornu samples have been taken from original animals and collected from commercial herbal medicine markets. 153 COI sequences have been successfully obtained from 155 samples through DNA extraction, PCR amplification, bidirectional sequencing and assembly. 93 COI sequences have been added to the DNA barcoding database of traditional Chinese animal medicine after validation using DNA barcoding GAP and tree-based methods. The species identification of the 62 commercial Bubali cornu medicines has been accomplished on the DNA barcoding system for identifying herbal medicine using the updated animal medicine database（www.tcmbarcode.cn）. Except two samples failed to obtain COI sequences, 54.8% of the commercial Bubali cornu medicines were water buffalo horns and 29% were yak horns. Our results showed that yak horn was the major adulterant of Bubali cornu and the DNA barcoding method may accurately discriminate Bubali cornu and their adulterants. Therefore, we recommend that supervision on the herbal medicine markets should be strengthened with this new method to warren the effectiveness of herbal medicines.

关 键 词：水牛角 COI序列 DNA条形码 鉴定 混伪品 

分 类 号：R931[医药卫生—生药学]