摘要
为研究DNA条形码技术在粉末类药材的应用,本文以市售鹿茸粉为对象,对其基原物种进行鉴定。依据2010版《中国药典》第三增补本中动物药材DNA条形码分子鉴定标准操作流程获得COI序列。对于测序峰图杂合度较高,可能为混合物的鹿茸粉,采用分子克隆方法得到序列。结果表明,利用PCR产物直接测序获得鹿茸粉的65条COI序列,其中有38%待检样品基原为2010版《中国药典》规定的梅花鹿Cervus nippon Temminck或马鹿Cervus elaphus Linnaeus,而62%为其他基原,以驯鹿Rangifer tarandus Linnaeus为主。进一步分析显示,不同价格、不同地域、不同公司抽检的样品或多或少存在非《中国药典》规定基原。利用分子克隆方法成功获得混合物样品的36条COI序列,主要基原为马鹿和梅花鹿。此外,有些待检样品包装明确标示为梅花鹿鹿茸粉,但鉴定结果为马鹿或驯鹿。因此,本研究证实DNA条形码技术可准确、有效鉴别市售鹿茸粉,为保障临床用药安全和市场监管提供新的技术手段,也为粉末类药材和饮片的鉴定提供示范。
In order to authenticate the components of antler powder in the market, DNA barcoding technology coupled with cloning method were used. Cytochrome c oxidase subunit 1 （COI） sequences were obtained according to the DNA barcoding components, the cloning method obtained from commercial antler standard operation procedure （SOP） was used to get each COl sequence. powders via sequencing PCR products For antler powder with possible mixed 65 COI sequences were successfully The results indicates that only 38% of these samples were derived from Cervus nippon Temminck or Cervus elaphus Linnaeus which is recorded in the 2010 edition of ＂Chinese Pharmacopoeia＂, while 62% of them were derived from other species. Rangifer tarandus Linnaeus was the most frequent species among the adulterants. Further analysis showed that some samples collected from different regions, companies and prices, contained adulterants. Analysis of 36 CO1 sequences obtained by the cloning method showed that C. elaphus and C. nippon were main components. In addition, some samples were marked clearly as antler powder on the label, however, C. elaphus or R. tarandus were their main components. In summary, DNA barcoding can accurately and efficiently distinguish the exactcontent in the commercial antler powder, which provides a new technique to ensure clinical safety and improve quality control of Chinese traditional medicine.
出处
《药学学报》
CAS
CSCD
北大核心
2015年第10期1356-1361,共6页
Acta Pharmaceutica Sinica
基金
重大新药创制国家科技重大专项"中药新药安全性检测技术与标准研究"(2014ZX09304307001)
关键词
鹿茸粉
DNA条形码
克隆
鉴定
antler powder
DNA barcoding
clone
identification
作者简介
通讯作者Tel：86—10—57833199，Fax：86—10—57833038，E-mail：slchen@implad．ac．cn；
通讯作者jysong@implad．ac．cn
