Research progress in original species identification in industry chain of Rhei Radix et Rhizoma / 中国中药杂志

Rhei Radix et Rhizoma is a kind of commonly used Chinese medicinal materials. Due to the overharvesting, the wild resource is endangering. Large market demand caused severely adulterant of commercial Rhei Radix et Rhizoma medicinal materials and decoction pieces. This manuscript reviewed the advances of the original species authentication in the industrial chain of Rhei Radix et Rhizoma during the latest decade, including characteristics and microscopic features, phytochemical analysis on anthraquinones, and molecular authentication based on DNA barcoding. Accordingly, an original species authentication route for the industrial chain of Rhei Radix et Rhizoma was summarized:(1)the identification of seeds and seedlings by DNA barcoding;(2) the selection of high variable sites based on the chloroplast genome;(3)biomonitoring of the Rhei Radix et Rhizoma medicinal materials and decoction pieces by two-dimensional DNA barcode;(4)traceability of Chinese patent medicines by third-generation sequencing. In conclusion, the combination of molecular identification and traditional identification methods provides a new idea for the identification of the original species of Rhei Radix et Rhizoma in the industrial chain and a essential guidance for the research of drug safety and efficacy of Rhei Radix et Rhizoma.

Analysis of Zingiberaceae in E'mei Area Using ITS2 Sequences / 中国实验方剂学杂志

Objective: To evaluate the use of ITS2 sequences as DNA barcode to identify the Zingiberaceae medicinal plants from E'mei area. Method: The genomic DNAs were extracted from 43 Zingiberaceae medicinal plant samples from Sichuan E'mei area. The ITS2 sequences of these samples were amplified and bidirectionally sequenced by PCR. 40 ITS2 sequences were downloaded from the GenBank,and then the interspecific and intraspecific genetic distances were calculated and analyzed by using MEGA 6.0 to construct Neighbor-joining (NJ) tree; TAXON DNA software was also used to analyze intraspecific and interspecific variations and barcoding gaps. The differences in secondary structure of the ITS2 sequences were predicted and compared. Result: The minimum interspecific distance in Zingiberaceae samples was greater than the maximum intra specific distance,with obvious barcoding gap. The NJ tree showed that the samples were clustered into five different branches,Alpinia,Curcuma,Globba,Hedychium,and Zingiber respectively,and further cluster into sub-branches. Significant differences were also present in the secondary structures of ITS2 between different samples. Conclusion: ITS2 sequences as DNA barcode can be used to conduct accurate and rapid identification of the Zingiberaceae plants and clearly figure out the phylogenetic relationship among them,providing guidance for the study of the distribution of medicinal plants of this genus,as well as theoretical basis for the quality control,medication safety and rational development of Zingiberaceae medicinal plants in E'mei area.

Textual Research on Name and Base Resource of Bangna-Tiebangchui / 中国中医药信息杂志

Objective To provide basis for the traceability and safty of Bangna-Tiebangchui according to textual research on the name and base resource of Bangna-Tiebangchui. Methods On the basis of literature textual research, combining herbal and modern Chinese (Tibetan) medicine specifications, literature research, medicinal name interpretation method and plant classification method were used for comprehensive analysis. Results Si Bu Yi Dian records toxic herb medicine of Banganabao. Jing Zhu Ben Cao divides Banganabao into five kinds with different types of efficacy according toxicity and colors, and records Langqingqietu (Tiebangchui) with similar toxicity from Yin Mountain or Han Areas. Base resource of Banganabao includes ten kinds of ranunculaceae aconitum plants. Conclusion Bangna (????? Tibetan transliteration) is the most toxic kind of Banganabao according to textual research, which is also named as Tiebangchui (TCM name) now. Bangna is widely used in anti-inflammation, analgesia, and local anesthesia, which base resource is Aconitum pendulum Busch and dry roots of A. flavum Hand.-Mazz.

Content Determination of Three Alkaloids in Aconitum Pendulum and Habitually Medicinal Materials / 中国中医药信息杂志

Objective To establish a method for determining benzoylaconine, aconitine and 3-deoxyaconitine in Aconitum pendulum and the habitually medicinal materials by HPLC; To provide basis for control of quality standard toxicity composition in Aconitum pendulum and difference between Aconitum pendulum and the habitually medicinal materials. Methods Agilent Eclipse XDB-C18 column (250 mm × 4.6 mm,5 μm) was used at 25 ℃ with the mobile phase consisting of acetonitrile-0.04% trimethylamine (each 1000 mL of water plus 4 mL triethylamine and 1.68 mL phosphoric acid) by gradient elution; detection wavelength was 235 nm; the flow rate was 1 mL/min; injection volume was 10 μL. Results The benzoylaconine, aconitine and 3-deoxyaconitine had good separation and linear relationship in the corresponding range (r＞0.999). The average recovery rates were 97.66%–98.47%, and RSD were 0.84%–1.60%. Conclusion The contents of 3 alkaloid were different in Aconitum pendulum and the habitually medicinal materials. Both A. polyschistum and A. sessiliflorum need separate drug names.

ITS2 sequence analysis and identification of medicinal Artemisia plants / 中国中药杂志

Artemisia hedinii occupies an important position in the Tibetan medicine. Plants in Artemisia vary a lot and are widely distributed in the Qinghai-Tibet Plateau, many plants in Artemisia look similar, making traditional identification methods laborious. In this article, ITS2 sequences were used as DNA barcoding to identify four kinds of confusable Tibetan medicine plants in Artemisia, aiming to establish a rapid and accurate identification methods. Twenty-one samples in Artemisia were collected from the Qinghai-Tibet Plateau, ITS2 sequence PCR amplification and sequencing were conducted after the extraction of DNA. Another 11 sequence downloaded from Genbank were added to the analysis. Genetic distance calculation and analysis, building Neighbor Joining (NJ) phylogenetic tree were conducted by MEGA 6.0, also comparison of secondary structures of ITS2 sequences among samples. A. hedinii, A. annua, A. dubia and A. argyi shared close genetic distance, but the maximum distance between the four species was much greater than the minimum distance within each species, NJ tree showed that the four species went to four separate branches, differences among secondary structures of ITS2 sequences also made it clear to identify these medical plants. It could be an accurate and rapid method for identification and recognition, as well as the evolutionary relationships between the species by using ITS2 sequence as DNA barcode for plants of Tibetan Artemisia. The study provides theoretical basis for quality control, medication safety and rational exploitation.

Identification and analysis of Corydalis boweri, Meconopsis horridula and their close related species of the same genus by using ITS2 DNA barcode / 中国中药杂志

The study is aimed to ensure the quality and safety of medicinal plants by using ITS2 DNA barcode technology to identify Corydalis boweri, Meconopsis horridula and their close related species. The DNA of 13 herb samples including C. boweri and M. horridula from Lhasa of Tibet was extracted, ITS PCR were amplified and sequenced. Both assembled and web downloaded 71 ITS2 sequences were removed of 5. 8S and 28S. Multiple sequence alignment was completed and the intraspecific and interspecific genetic distances were calculated by MEGA 5.0, while the neighbor-joining phylogenetic trees were constructed. We also predicted the ITS2 secondary structure of C. boweri, M. horridula and their close related species. The results showed that ITS2 as DNA barcode was able to identify C. boweri, M. horridula as well as well as their close related species effectively. The established based on ITS2 barcode method provides the regular and safe detection technology for identification of C. boweri, M. horridula and their close related species, adulterants and counterfeits, in order to ensure their quality control, safe medication, reasonable development and utilization.

Content comparison of buddleoside and pectolinarin in Cirsium japonicum, C. leo and C. leducei / 中国中药杂志

<p><b>OBJECTIVE</b>To compare Cirsium japonicum characteristics with C. leo and C. leducei, along with the content of buddleoside and pectolinarin, and lay the foundation for the quality control of C. japonicum.</p><p><b>METHOD</b>Samples were collected and the relevant drugs were bought. The samples were divided into root, stem, leaf and flower, and the content of buddleoside and pectolinarin was determine by the HPLC. Chromatographic column: Waters XBridge C18 (4.6 mm x 250 mm), mobile phase: methanol-water (45: 55), measurement wavelength: 326 nm, flow rate: 0.8 mL x min(-1), column temperature: 30 degrees C. RESULT AND CONDUSION: Standard curve equation of buddleoside: Y = 74 064X-47 748, R2 = 0.991. Standard curve equation of pectolinarin: Y = 1 711 64X - 180 707, R2 = 0.999. The content of buddleoside: C. japonicum leaf was 1.987 3%, C. leo leaf 1.412 2%, C. leducei leaf 0.149 2%. The content of buddleoside was lower in root and stem. Pectolinarin was not detected in the C. japonicum and C. leo. The pectolinarin content was 0.069 0% in C. leducei leaf.</p>

HPLC fingerprint of Rhizoma Polygoni Cuspidati / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the HPLC fingerprint of Rhizoma Polygoni Cuspidati (Polygonum cuspidatum).</p><p><b>METHOD</b>The HPLC separation was carried with Diamonsil C18 column and eluted with a gradient from methanol and 0.1% phosphoric acid, the detection wavelength was at 230 nm and recording 70 min. The similarity of chromatograms was compared by mean of the software from Zhongnan University.</p><p><b>RESULT</b>The constituents of P. cuspidatum were well separated by HPLC, and the similarity was above 0.80.</p><p><b>CONCLUSION</b>The method can be used for the study of fingerprints of Rhizoma Polygoni Cuspidati.</p>