Identification of <italic style="font-style: italic">Levisticum officinale </italic>Adulterated in <italic style="font-style: italic">Angelica sinensis</italic> by PCR-RFLP

Zhong-fei SHI; Bao-xia TENG; Jing LAI; Lin NI; Ping-shun SONG

doi:10.13422/j.cnki.syfjx.20210514

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Identification of Levisticum officinale Adulterated in Angelica sinensis by PCR-RFLP

Pharmacy Fundamentals | 更新时间：2021-04-02

- Identification of Levisticum officinale Adulterated in Angelica sinensis by PCR-RFLP
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 27, Issue 9, Pages: 168-175(2021)
- 作者机构：
  
  甘肃省药品检验研究院国家中药材及饮片质量控制重点实验室，甘肃省中藏药检验检测技术工程实验室，兰州 730000
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20210514
  CLC： R284.2;R289;R22;R2-031
- Received：23 November 2020，
  
  Published Online：22 February 2021，
  
  Published：05 May 2021
- 稿件说明：
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史中飞,滕宝霞,赖晶等.PCR-RFLP鉴别当归药材及饮片中掺混伪品——欧当归的方法[J].中国实验方剂学杂志,2021,27(09):168-175.

SHI Zhong-fei,TENG Bao-xia,LAI Jing,et al.Identification of Levisticum officinale Adulterated in Angelica sinensis by PCR-RFLP[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(09):168-175.
史中飞,滕宝霞,赖晶等.PCR-RFLP鉴别当归药材及饮片中掺混伪品——欧当归的方法[J].中国实验方剂学杂志,2021,27(09):168-175. DOI： 10.13422/j.cnki.syfjx.20210514.

SHI Zhong-fei,TENG Bao-xia,LAI Jing,et al.Identification of Levisticum officinale Adulterated in Angelica sinensis by PCR-RFLP[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(09):168-175. DOI： 10.13422/j.cnki.syfjx.20210514.

摘要

目的

该研究拟通过限制性片段长度多态性聚合酶链反应方法（PCR-RFLP）建立一种能快速鉴别当归药材及饮片中掺伪欧当归的方法。

方法

通过比对当归和欧当归的核糖体DNA内转录间隔区（ITS）序列酶切位点，选择欧当归的特异性酶切位点Fnu4HⅠ，并设计PCR-RFLP反应引物，对影响PCR-RFLP反应的退火温度、引物浓度、循环数和酶切时间等条件进行优化，并对该方法的准确性进行了考察。另外，利用建立的PCR-RFLP鉴别方法对掺伪不同比例和不同来源的当归掺伪样品进行了适用性考察。

结果

建立了当归药材及饮片中掺伪欧当归的PCR-RFLP鉴别方法，在退火温度63 ℃，循环数为30时，掺有欧当归的当归样品经过特异性引物扩增后，能被Fnu4HⅠ限制性内切酶酶切，在100~500 bp检出2条单一DNA条带，当归样品则无此条带。该方法对当归中掺入欧当归的检出限为3%，可用于日常当归中掺混欧当归的检测。

结论

建立的PCR-RFLP鉴别方法能够灵敏、准确地检测当归药材及饮片中是否掺有欧当归，可为当归的质量控制提供检验依据，以保障其临床用药安全。

Abstract

Objective

To establish a rapid method to identify

Levisticum officinale

adulterated in

Angelica sinensis

by polymerase chain reaction -restriction fragment length polymophism（PCR-RFLP）.

Method

By comparing sequences restriction sites in ribosomal DNA Internal Transcribed Spacer（ITS） of

A. sinensis

and

L. officinale

，the specific restriction site Fnu4HI of

L. officinale

was selected，and the primers for PCR-RFLP reaction were designed. Different

A. sinensis

and

L. officinale

were amplified by PCR. The conditions affecting the PCR-RFLP reaction，such as annealing temperature，primer concentration，cycle number and enzyme digestion reaction time，were optimized，and the accuracy of the method was investigated. The established PCR-RFLP identification method was used to investigate the applicability of

L. officinale

adulterated

in A. sinensis

with different aduleration ratios and different origins.

Result

A PCR-RFLP method for identifying

A. sinensis

mixed with

L. officinale

was established. When the annealing temperature was 62 ℃ and the number of cycles was 30，when the

L. officinale

adulterated in

A. sinensis

could be digested by Fnu4H I restriction endonuclease after amplification with specific primers，and the two single DNA bands were detected between 100-500 bp，the

A. sinensis

were all negative. The minimum detection limit of this method for adulterated

L. officinale

A. sinensis

was 3%，which could be used for the detection of adulterated

L. officinale

A. sinensis

Conclusion

The established PCR-RFLP identification method is sensitive and accurate in detecting whether there is

L. officinale

A. sinensis

，and it provides inspection reference and basis for the quality control of

A. sinensis

，with great significance to ensure the safety of its clinical medication.

关键词

Keywords

references

国家药典委员会 . 中华人民共和国药典：一部［M］. 北京：中国医药科技出版社， 2020 ： 139 .

史巧霞，杨锡仓 . 欧当归的合理开发与应用探讨［J］. 内蒙古中医药， 2016 ， 35 （ 16 ）： 37 .

胡双丰 . 欧当归不可代替中药当归药用［J］. 中国当代医药， 2009 ， 16 （ 4 ）： 41 - 45 .

方洪钜，吕瑞绵，刘国声，等 . 挥发油成分的研究——Ⅱ.中国当归与欧当归主要成分的比较［J］. 药学学报， 1979 （ 10 ）： 617 - 623 .

崔占虎，龙平，王颖莉，等 . DNA分子标记技术在中成药鉴定中的应用与展望［J］. 中药材， 2015 ， 38 （ 1 ）： 188 - 192 .

杨慧洁，杨世海，张淑丽，等 . 药用植物DNA条形码研究进展［J］. 中草药， 2014 ， 45 （ 18 ）： 2581 - 2587 .

梁宇庭，蒋超，周骏辉，等 . 高卢蜜环菌PCR-RFLP快速鉴别方法研究［J］. 中国中药杂志， 2019 ， 44 （ 17 ）： 3622 - 3626 ..

WANG C Z ， LI P ， DING J Y ， et al . Identification of Fritillaria pallidiflora using diagnostic PCR and PCR-RFLP based on nuclear ribosomal DNA internal transcribed spacer sequences ［J］. Planta Med ， 2005 ， 71 （ 4 ）： 384 - 386 .

WANG C Z ， LI P ， DING J Y ， et al . Simultaneous identification of Bulbus Fritillariae cirrhosae using PCR-RFLP analysis ［J］. Phytomedicine ， 2006 ， 14 （ 9 ）： 628 - 632 .

张婷，徐珞珊，王峥涛，等 . 药用植物束花石斛、流苏石斛及其形态相似种的PCR-RFLP鉴别研究［J］. 药学学报， 2005 ， 40 （ 8 ）： 728 - 733 .

王凤霞，陈媛媛，任贵奇，等 . 鹿血的PCR-RFLP鉴定研究［J］. 中草药， 2018 ， 49 （ 8 ）： 1914 - 1918 .

李谦，刘杰，过立农，等 . 基于ITS2序列的紫草PCR-RFLP鉴别研究［J］. 药物分析杂志， 2016 ， 36 （ 9 ）： 1611 - 1617 .

温子帅，刘爱朋，景松松，等 . 当归和欧当归的定性与定量鉴别［J］. 中成药， 2018 ， 40 （ 12 ）： 2719 - 2723 .

辛天怡，姚辉，韩建萍，等 . 当归药材及其混伪品ITS2条形码鉴定［J］. 科研信息化技术与应用， 2013 ， 4 （ 3 ）： 76 - 82 .

刘阳，杨淑霞，李敏惠，等 . 引物浓度与退火温度不当导致巢式PCR非特异性扩增［J］. 成都医学院学报， 2008 （ 2 ）： 111 - 114 .

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Identification of Levisticum officinale Adulterated in Angelica sinensis by PCR-RFLP

DOI：10.13422/j.cnki.syfjx.20210514

摘要

Abstract

关键词

Keywords

references