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铁皮石斛的ITS2条形码分子鉴定及5种重金属及有害元素的测定
  • ISSN号:0254-1793
  • 期刊名称:《药物分析杂志》
  • 时间:0
  • 分类:R917[医药卫生—药物分析学;医药卫生—药学]
  • 作者机构:[1]北京中医药大学,北京100102, [2]中国食品药品检定研究院,北京100050, [3]中国医学科学院北京协和医学院,药用植物研究所濒危药材繁育国家工程实验室,北京100193
  • 相关基金:国家十二五“重大新药创制”专项(2014ZX09304307-002); 国家自然科学基金青年项目(81274025)
中文摘要:

目的:应用DNA条形码技术,从分子水平快速、准确地鉴定我国12个省区铁皮石斛栽培基地铁皮石斛(Dendrobium officinale Kimura et Migo)及其同属常见植物的基原,采用微波消解ICP-MS技术测定各地铁皮石斛中铅(Pb)、镉(Cd)、砷(As)、汞(Hg)、铜(Cu)5种重金属及有害元素的含量,评价栽培铁皮石斛的用药安全。方法:提取各样本总DNA,对r DNA的第二内转录间隔区(ITS2区)进行扩增,计算种内、种间Kimura 2-parameter(K2P)遗传距离,通过与中药材DNA条形码鉴定系统比对和构建N-J(Neighbor-Joining)系统树进行基原鉴定。建立微波消解方法进行样品前处理,ICP-MS法对铁皮石斛中5种重金属及有害元素进行测定,比对国内外现行标准,对铁皮石斛的安全性进行评价。结果:铁皮石斛的ITS2序列长度在246-340bp范围,种内变异较小,种内最大K2P遗传距离为0.009,铁皮石斛及其混伪品的种间平均K2P遗传距离为0.206,种间最小K2P遗传距离明显大于种内的最大K2P遗传距离。经与中药材DNA条形码鉴定系统比对及构建N-J树进行基原鉴定。结果表明ITS2序列可区分铁皮石斛及其近缘种。5种重金属及有害元素的测定结果表明各地的含量差异较显著,但在安全的范围内。结论:ITS2片段DNA条形码可有效鉴定中药铁皮石斛及其常见同属植物的基原,建立的ICP-MS法测定铁皮石斛中5种重金属与有害元素的方法准确、可行,各元素的含量在现行相关标准的安全范围内。

英文摘要:

Objective:To identify Dendrobium officinale and its common affinitive species on a molecule level by using ITS2 barcoding technique,and to evaluate the medication safety of Dendrobium officinale from 12 provinces in China through detection of five heavy metals and hazardous elements of Pb,Cd,As,Hg,and Cu by microwave digestion and ICP-MS technique.Methods:Total genomic DNA was extracted from Dendrobium officinale.The internal transcribed spacer 2(ITS2) regions were amplified using polymerase chain reaction(PCR),and PCR products were sequenced bi-directionally.The Kimura 2-parameter(K2P) distances and nucleotide composition of GC content were computed.Alignment with the known sequence from Gen Bank and species identification analyses were conducted through the species identification system of neighbor-joining(N-J) trees.The contents of 5 heavy metals and hazardous elements in Dendrobii Officinalis Caulis were determined successfully.The safety was evaluated by comparison of the domestic and foreign correlated standards.Results:The ITS2 sequence lengths of Dendrobium officinale were within 246-340 bp,and the intra-species variation was small.The maximum intra-specific genetic distances of Dendrobium officinale was 0.009.The average inter-specific genetic distances between Dendrobium officinale and analogues species were 0.206.The results showed that the minimum inter-specific divergence was larger than the maximum intra-specific divergence.The results of the species identification system for traditional Chinese medicine by DNA barcodes and N-J trees indicated that Dendrobium officinale and the affinitive species can be clearly identified.The contents of five heavy metals and hazardous elements were obviously different in Dendrobii Officinalis Caulis samples from different producing places,but all in accordance with the limits of domestic and foreign correlative standards.Conclusion:DNA barcodes can accurately identify the origin of Dendrobium officinale and its common affinitive species.The

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期刊信息
  • 《药物分析杂志》
  • 中国科技核心期刊
  • 主管单位:中国科协
  • 主办单位:中国药学会
  • 主编:金少鸿
  • 地址:北京天坛西里2号
  • 邮编:100050
  • 邮箱:ywfx@nifdc.org.cn
  • 电话:010-67058427
  • 国际标准刊号:ISSN:0254-1793
  • 国内统一刊号:ISSN:11-2224/R
  • 邮发代号:2-237
  • 获奖情况:
  • 四通杯全优期刊奖,92、97年获科协优秀期刊三等奖,96年影响因子全年第一
  • 国内外数据库收录:
  • 美国国际药学文摘,美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国英国皇家化学学会文摘,中国北大核心期刊(2000版)
  • 被引量:32603