Forensically informative nucleotide sequencing (FINS) for the authentication of Chinese medicinal materials

Chinese medicinal materials may be authenticated by molecular identification. As a definitive approach to molecular identification of medicinal materials, forensically informative nucleotide sequencing (FINS) comprises four steps, namely (1) DNA extraction from biological samples, (2) selection and amplification of a specific DNA fragment, (3) determination of the sequence of the amplified DNA fragment and (4) cladistic analysis of the sample DNA sequence against a DNA database. Success of the FINS identification depends on the selection of DNA region and reference species. This article describes the techniques and applications of FINS for authenticating Chinese medicinal materials. © 2011 Li et al; licensee BioMed Central Ltd.published_or_final_versio

Current approaches for the authentication of medicinal Dendrobium species and its products

Dendrobium Sw., a member of the family Orchidaceae, includes approximately 1100 species distributed in different parts of the world. In China, the genus is represented by 76 species and two varieties, of which D. loddigesii, D. fimbriatum, D. chrysanthum, D. officinale (=D. candidum) and D. nobile are listed in the Chinese Pharmacopoeia as the source material for Herba Dendrobii (Shihu). Because of increased demand and high price, Herba Dendrobii is often adulterated in the trade by other related species. Many Dendrobium species are over-collected from the wild and are listed under endangered taxa in the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). Therefore, a reliable authentication method is needed to regulate the trade and help conserve the species from unscrupulous collections. This review describes the present status of using medicinal Dendrobium in China, current approaches to authenticate medicinal Dendrobium plants and Herba Dendrobii, and the efforts under way towards the development of DNA microarrays to facilitate differentiation/identification of genuine material in complex Chinese medicinal formulations. © NIAB 2005.link_to_subscribed_fulltex

DNA Microarray for Identification of the Herb of Dendrobium Species from Chinese Medicinal Formulations

A DNA microarray for detecting processed medicinal Dendrobium species (Herba Dendrobii) was constructed by incorporating the ITS1-5.8S-ITS2 sequences of 16 Dendrobium species on a glass slide. Using fluorescence-labeled ITS2 sequences as probes, distinctive signals were obtained for the five medicinal Dendrobium species listed in the Chinese Pharmacopoeia. The established microarray was able to detect the presence of D. nobile in a Chinese medicinal formulation containing nine herbal components.link_to_subscribed_fulltex

A DNA microarray for differentiation of the Chinese medicinal herb Dendrobium officinale (Fengdou Shihu) by its 5 S ribosomal DNA intergenic spacer region

A DNA microarray was constructed for high-throughput identification of the plant resource of commercial FDSH [Fengdu Shihu (Dendrobium officinale)]. The 5 S rDNA (ribosomal DNA) intergenic spacer region in D. officinale, D. nobile, D. moniliforme, D. hercoglossum, D. williamsonii, D. capillipes, D. wilsonii and D. jenkinsii was amplified by a single primer pair and sequenced. The sequences showed polymorphism. They were incorporated on a glass slide and hybridized with fluorescently labelled 5 S sequences from commercial Shihu. The DNA microarray enabled the differentiation of D. officinale from the other species tested. FDSH could thus be distinguished from its adulterants. It is evident that DNA microarrays provide a high-throughput and reliable approach for the identification of plant resources, and the method presented here is useful for the authentication of FDSH. © 2008 Portland Press Ltd.link_to_subscribed_fulltex

Differentiation of medicinal Codonopsis species from adulterants by polymerase chain reaction-restriction fragment length polymorphism

DNA sequence analysis of rDNA internal transcribed spacer (ITS) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were exploited for their applications in differentiating medicinal species Codonopsis pilosula, C.tangshen, C.modesta, and C.nervosa var. macrantha, from two related adulterants Campanumoeo javania and Platycodon grandiflorus. The data demonstrated that the rDNA ITSI and ITSII sequences of the four Codonopsis are highly homologous but not identical, and are significantly different from those of the two adulterants. The sequence difference allows effective and reliable differentiation of Codonopsis from the adulterants by PCR-RFLP

Sesquiterpene lactones from Elephantopus scaber

The whole plant of Elephantopus scaber afforded the known deoxyelephantopin and isodeoxyelephantopin, and a new germacranolide sesquiterpene lactone named scabertopin, whose structure and stereochemistry were determined by spectroscopic methods and single-crystal X-ray analysis. Copyright (C) 1996 Elsevier Science Lt

Random primed polymerase chain reaction differentiates Codonopsis pilosula from different localities

DNA fingerprints distinctive among the samples from different localities in China were successfully reproduced for the Chinese herb Dangshen, the roots of Codonopsis pilosula, (Campanulaceae). Similarity index (S.I.) analysis revealed that C. pilosula samples from the same province generated similar DNA fingerprints, while samples of different provinces displayed different DNA fingerprints. This method may be a general and valuable tool for locality authentication of other Chinese herbal medicinal materials.link_to_subscribed_fulltex

Chemical and molecular characterization of Hong Dangshen, a unique medicinal material for diarrhea in Hong Kong

Aim: To investigate the plant origin and the identity of the red substance on the surface of Hong Dangshen, a unique medicinal material for diarrhea in Hong Kong. Methods The HPLC fingerprints and 5S rRNA gene spacer sequences of Hong Dangshen were obtained and compared with those of genuine species of Radix Codonopsis. The X-ray diffraction spectrum of the red substance was analyzed and compared with that of Halloysitum Rubrum. Results Hong Dangshen showed very similar HPLC fingerprints to the Codonopsis species and the highest similarity to Codonopsis pilosula var. modesta in terms of 5S rRNA gene spacer sequences. The X-ray diffraction spectrum of the red substance was consistent with that of Halloysitum Rubrum. Conclusion The source plant of Hong Dangshen was suggested to be Codonopsis pilosula var. modesta, one of the genuine original plants of Radix Codonopsis (Dangshen) in the China Pharmacopoeia (2005 edition). The red substance on the surface of Hong Dangshen was indicated to be Halloysitum Rubrum, a traditional medicinal mineral for chronic diarrhea. Our data suggest that Hong Dangshen is derived from the roots of Codonopsis pilosula var. modesta which has been processed with Halloysitum Rubrum, and the name is suggested to be Radix Codonopsis Praeparata Halloysita Rubra. 目的鑒定香港特有止瀉中藥材紅黨參的基源植物和表面紅色物質。方法將紅黨參的5SrRNA基因間隔區序列和HPLC色譜指紋譜與正品黨參5SrRNA基因間隔區序列和HPLC色譜指紋譜進行比較研究;將紅黨參表面紅色物質的X-衍射光譜與中藥赤石脂X-衍射光譜進行比較研究。結果紅黨參的HPLC色譜指紋譜和黨參藥材具有相同的特征色譜峰;紅黨參的5SrRNA基因間隔區序列與黨參正品之一的素花黨參(Codonopsis pilosula var.modesta)的5SrRNA基因間隔區序列最為相似;紅色物質的X-衍射光譜與中藥赤石脂X-衍射光譜完全相同。結論紅黨參的基源是素花黨參(Codonopsis pilosula var.modesta)的根,紅黨參表面的紅色物質是傳統中藥赤石脂(Halloysitum Rubrum)。紅黨參是素花黨參根用中藥赤石脂炮制而成,科學名可定為Radix Codonopsis Praeparata Halloysita Rubra