论中药分子鉴定的方法和原则

doi:10.3724/SP.J.1143.2012.12162

Plant Diversity ›› 2012, Vol. 34 ›› Issue (6): 607-613.DOI: 10.3724/SP.J.1143.2012.12162

论中药分子鉴定的方法和原则

袁庆军1, 张文婧1、2, 姜丹1、2, 张永清2, 马超一1, 黄璐琦1

1 中国中医科学院中药研究所，北京100700; 2 山东中医药大学药学院，山东济南250355

收稿日期:2012-11-20 出版日期:2012-12-25 发布日期:2012-11-28
基金资助:
国家科技部科技基础工作专项项目；国家高科技研究发展计划 (863计划) (2012AA021801)；中国科学院大科学装置开放研究项目 (2009-LSFGBOWS-01)；国家自然科学基金重点项目 (81130070)；中国中医科学院中药研究所基本科研业务费自主选题项目 (2011ADXK-01)

On the Methods and Principles of Molecular Identification of Chinese Herbs

YUAN Qing-Jun-1, ZHANG Wen-Jing-1、2, JIANG Dan-1、2, ZHANG Yong-Qing-2, MA Chao-Yi-1, HUANG Lu-Qi-1

1 Institute of Chinese Materia Medica, China Academy of Chinese Medicinal Sciences, Beijing 100700, China;
2 College of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China

Received:2012-11-20 Online:2012-12-25 Published:2012-11-28

摘要/Abstract

摘要：

中药的准确鉴定涉及到人民生命安全和切身利益。传统的鉴定方法，即感官评价、显微鉴定和理化鉴定均存在不同程度的局限性，而分子鉴定则为中药的快速和准确鉴定带来了新的契机。为了使中药的分子鉴定得到更加广泛和有效的应用，应该高度重视相关标准、规范的制订。本文提出了中药分子鉴定的一些主要方法：1)真伪品鉴定：特异聚合酶链式反应法；2)正品和替代品鉴定：DNA条形码鉴定法；3)多基源鉴定：群体遗传学分析法；4)产地鉴别：分子谱系地理学分析法。经上述方法仍无法鉴别的贵重药材可进一步应用人类亲子鉴定的方法，开发特异微卫星标记进行进一步的鉴定。

关键词: 中药, 分子鉴定, 真伪品, 正品和替代品, 多基源, 产地鉴别

Abstract:

Accurate identification of herbal medicinal materials is relevant to the safety of human life and economic interests. Traditional identification methods, including sensory evaluation, microscopic identification, physical and chemical identification, all have their limitations. Molecular identification brings a new opportunity for accurate identification of herbal medicinal materials. However, prior to its wide adoption, the methods and principles of molecular identification should be fully discussed. In this paper, we proposed a set of new methods for molecular authentication of herbal medicinal materials: 1) Identification between authenticity and adulteration by using specific polymerase chain reaction; 2) Identification between official herb and substitute by using the method of DNA barcoding; 3) Identification among multiple species of one official herbs by constructing genealogy among closelyrelated species based on population genetics; 4) Identification among herbs of different geographical origins by phylogeographybased analysis. For those that can not be identified by above four methods, more rapidly evolved markers such as microsatellite should be employed and individualbased analysis could be adopted.

Key words: Herbal medicines, Molecular identification, Authenticity and adulteration, Official herb and substitute, Multiple species origin, Different regional origins

中图分类号:

袁庆军1, 张文婧1、2, 姜丹1、2, 张永清2, 马超一1, 黄璐琦1. 论中药分子鉴定的方法和原则[J]. Plant Diversity, 2012, 34(6): 607-613.

YUAN Qing-Jun-, ZHANG Wen-Jing-, JIANG Dan-, ZHANG Yong-Qing-, MA Chao-Yi-, HUANG Lu-Qi. On the Methods and Principles of Molecular Identification of Chinese Herbs[J]. Plant Diversity, 2012, 34(6): 607-613.

参考文献

陈士林, 2012. 中药DNA条形码分子鉴定［M］. 北京: 人民卫生出版社
国家药典委员会, 2010. 中华人民共和国药典［M］. 北京: 中国医药科技出版社, 72, 349
黄璐琦, 2000. 分子生药学［M］. 北京: 北京大学医学出版社
刘晓光, 2012. 基于群体遗传学的黄连属DNA条形码研究［D］.［学位论文］. 北京: 中国中医科学院中药研究所, 济南: 山东中医药大学
邵鹏柱, 曹晖, 王骏等, 2004. 中药分子鉴定［M］. 上海: 复旦大学出版社
张彬, 2012. 当归属药用植物及药材的DNA条形码鉴别研究——兼论DNA条形码在中药材鉴定中的原则研究［D］.［学位论文］. 北京: 中国中医科学院中药研究所
Arnold M, 1997. Natural Hybridization and Evolution［M］. Oxford: Oxford University Press
Avise JC, Arnold J, Ball Jr. RM et al., 1987. Intraspecific phylogeography: The mitochondrial DNA bridge between population genetics and systematics［J］. Annual Review of Ecology and Systematics, 18: 489—522
Avise JC, 1996. Space and time as axes in intraspecific phylogeography［A］. In: Huntley B, Cramer W, Morgan AV et al. (eds.), Past and Future Rapid Environmental Change: The Spatial and Evolutionary Responses of Terrestrial Biota［M］. New York: SpringerVerlag, 381—388
Avise JC, 2000. Phylogeography: The History and Formation of Species ［M］. Cambridge, Massachuseets, London, England: Harvard University Press
Barrett RDH, Hebert PDN, 2005. Identifying spiders through DNA barcodes［J］. Canadian Journal of ZoologyRevue Canadienne De Zoologie, 83: 481—491
But PPH, 1994. Herbal poisoning caused by adulterants or erroneous substitutes［J］. Journal of Tropical Medicine and Hygiene, 97: 371—374
But PPH, Tomlinson B, Cheung KO et al., 1996. Adulerants of herbal products can cause poisoning［J］. British Medical Journal, 313: 117
CBOL Plant Working Group, 2009. A DNA barcode for land plants［J］. Proceedings of the National Academy of Sciences of the United States of America, 106: 12794—12797
Chase MW, Cowan RS, Hollingsworth PM et al., 2007. A proposal for a standardised protocol to barcode all land plants［J］. Taxon, 56: 295—299
Chen SL, Yao H, Han JP et al., 2010. Validation of the ITS2 region as a nowel DNA barcode for identifying medicinal plant species［J］. PloS One, 5: e8613
Chiang TY, Schaal BA, 1999, Phylogeography of North American populations of the moss species Hylocomium splendens based on the nucleotide sequence of internal transcribed spacer 2 of nuclear ribosomal DNA［J］. Molecular Ecology, 8: 1037—1042
Excoffier L, Smouse PE, 1994. Using alleles frequencies and geographic subdivision to reconstruct gene trees within a species: molecular variance parsimony［J］. Genetics, 136: 343—359
Fazekas AJ, Kesanakurti PR, Burgess KS et al., 2009. Are plant species inherently harder to discriminate than animal species using DNA barcoding markers?［J］. Molecular Ecology Resources, 9: 130—139
Feng T, Downie SR, Yan Y et al., 2009. Molecular systematics of Angelica and allied genera (Apiaceae) from the Hengduan Mountains of China based on nrDNA ITS sequences: phylogenetic affinities and biogeographic implications［J］. Journal of Plant Research, 122: 403—414
Futuyma DJ, 1997. Evolutionary Biology［M］. Massachusetts: Inc. Sunderland
Gao LM (高连明), Liu J (刘杰), Cai J (蔡杰) et al., 2012. A synopsis of technical notes on the standards for plant DNA barcoding［J］. Plant Diversity and Resources (植物分类与资源学报), 34 (6): 592—606
Hebert PDN, Cywinska A, Ball SL et al., 2003. Biological identifications through DNA barcodes［J］. Proceedings of the Royal Society of London Series B: Biological Sciences, 270: 313—321
Hebert PDN, Stoeckle MY, Zemlak TS et al., 2004. Identification of birds through DNA barcodes［J］. PLoS Biology, 2: e312
Huang LQ (黄璐琦), Zhang RX (张瑞贤), 1997. Discussion on the biology of geoherbs［J］. Chinese Pharmaceutical Journal (中国药学杂志), 32 (9): 563
Kress WJ, Erickson DL, 2007. A twolocus global DNA barcode for land plants: the coding rbcL gene complements the noncoding trnHpsbA spacer region［J］. PLoS One, 2: e508
Li DZ, Gao LM, Li HT et al., 2011. Comparative analysis of a large dataset indicates that ITS shoud be incorporated into the core barcode for seed plants［J］. Proceedings of the National Academy of Sciences of the United States of America, 108 (49): 19641—19646
Meyer CP, Paulay G, 2005. DNA barcoding: rrror rates based on comprehensive sampling［J］. PLoS Biology, 3 (12): 2229—2238
Petit RJ et al., 2003. Glacial refugia: hotspots but not melting post of genetic diversity［J］. Science, 300: 1563—1565
Schaal BA, Hayworth DA, Olsen KM et al., 1998. Phylogeographic studies in plants: problems and prospects［J］. Molecular Ecology, 7: 465—475
Templeton AR, 1998. Nested clade analyses of phylogeographic data: testing hypotheses about gene flow and population history［J］. Molecular Ecology, 7: 381—397
Vanhaelen M, VanhaelenFastre R, But PPH et al., 1994. Indentification of aristolochic acid in Chinese herbs［J］. Lancet, 343: 174
Vanherweghem JL, Delielreux M, Tielemans C et al., 1993. Rapidly progressive interstitial renal fibrosis in young women: Association with slimming regimen including Chinese herbs［J］. Lancet, 341: 387—391
World Health Organization, 2008. Traditional Medicine. Fact Sheet N134. Geneva: World Health Organization
Xiao XH (肖小河), Chen SL (陈士林), Huang LQ (黄璐琦) et al., 2009. Survey of investigations on Daodi Chinese medicinal materials in China since 1980s［J］. China Journal of Chinese Materia Medica (中国中药杂志), 34 (5): 519

[1]	王海坤1, 2 , 李涛1 , 赵丹丹3 , 李凌飞1 , 沙涛1 , 赵之伟1 . 两株滇产广义美味牛肝菌的分离培养及其分子鉴定[J]. Plant Diversity, 2007, 29(05): 559-562.
[2]	陈纪军张壮鑫周俊. 著名中药白首乌中的新c21甾体甙[J]. Plant Diversity, 1988, 10(04): 1-3.
[3]	孙汉董林中文钮芳娣. 伞形科中药的研究Ⅶ．白亮独活根的化学成分(1)[J]. Plant Diversity, 1984, 6(01): 1-3.
[4]	孙汉董林中文钮芳娣. 伞形科中药的研究（Ⅵ)长前胡的化学成分（第二报）[J]. Plant Diversity, 1983, 5(04): 1-3.
[5]	孙汉董;林中文;钮芳姊. 伞形科中药的研究Ⅴ.南瓜七根的化学成分[J]. Plant Diversity, 1982, 4(01): 1-3.
[6]	孙汉董;林中文;钮芳娣;了靖垲. 伞形科中药的研究 Ⅳ.法落海素的结构[J]. Plant Diversity, 1981, 3(03): 1-3.
[7]	孙汉董;林中文;钮芳娣;丁靖凯. 伞形科中药的研究 Ⅲ、长前胡的新香豆素[J]. Plant Diversity, 1981, 3(02): 1-3.
[8]	伍明珠. 滇产植物的皂素成份研究IV，IV中药三七的两种皂甙（1）[J]. Plant Diversity, 1979, 1(01): 1-3.

论中药分子鉴定的方法和原则

On the Methods and Principles of Molecular Identification of Chinese Herbs

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 8

编辑推荐

Metrics

本文评价