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OBJECTIVE：To improve the quality standard of Taraxaci Herba ，and to evaluate the quality of T. officinale from different origins. METHODS ：Based on the provisions of the 2020 edition of Chinese Pharmacopoeia （part Ⅰ）under the item “Taraxaci Herba ”，the method of content determination was added for the detection of water-soluble extracts （hot extraction method）and alcohol-soluble extracts （hot extraction method ），total flavonoids ，chlorogenic acid ，caffeic acid ，cichoric acid and isochlorogenic acid A. HPLC fingerprint was established by using 42 batches of T. officinale from 8 producing areas as object ，and principal component analysis was performed on the basis of above results. RESULTS ：The contents of alcohol-soluble extracts in 42 batches of T. officinale were 15.30％-30.40％，and those of water-soluble extracts were 27.59％-38.96％. The concentration of total flavonoids（UV spectrophotometry ），chlorogenic acid ，caffeic acid ，cichoric acid and isochlorogenic acid A （HPLC method ）were 0.016-0.096，0.003-0.196，0.004-0.117，0.025-1.578，0.002-0.152 mg/mL，respectively （all R2＞0.999）；RSDs of precision ， stability and repeatability tests were all lower than 2.00％（n＝6）；average sample recovery were 98.97％-103.53％，and RSDs were 1.19％-1.58％. The contents of total flavonoids ，chlorogenic acid ，caffeic acid ，cichoric acid and isochlorogenic acid A were 0.734％ -3.700％ ，0.004％ -0.123％ ，0.006％ - 0.087％ ，0.073％ -1.499％ ，0.005％ -0.109％ respectively in 42 batches of T. officinale. For 42 batches of T. officinale samples in HPLC fingerprint ，RSDs of the relative retention time of the common peak were 0-0.94％，and RSDs of the relative peak area were 0-125.57％. Among them ，the similarity of 39 batches of samples was all higher than 0.900. Results of principal component analysis showed that the quality of T. officinale from Shaanxi province was better，followed by medicinal materials from Hebei province. CONCLUSIONS ：Tentatively，the contents of alcohol-soluble extract，water-soluble extract ，total flavonoids ，chlorogenic acid ，caffeic acid ，cichoric acid and isochlorogenic acid A shall not be less than 17.0％，27.0％，1.383％，0.024％，0.021％，0.450％，0.021％ for Taraxaci Herba. In addition to the low content of caffeic acid in T. officinale from Shaanxi province ，the other indexes were better ；the content of caffeic acid in T. officinale from Hebei province was higher than that from Shaanxi province ，and other indicators were slightly lower than that from Shaanxi province. The quality of comprehensive evaluation of T. officinale from other origins was relatively poor ，and the quality of different batches of medicinal materials from the same origin was unstable.
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Objective To establish a Ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS) chromatographic method for Xiefei-Pingchuanling extract, and to analyze the main components of its water extract. Methods The UPLC-MS was adopted with acetonitrile -0.2% formic acid aqueous solution (gradient elution) as mobile phase at a flow rate of 0.8 ml/min, detection wave length at 254 nm, and column temperature was 30 ℃. Results Eighteen components were separated from Xiefei-Pingchuanling extract, and eight ingredients were identified, such as the ephedrine, pseudoephedrine hydrochloride, salvianolic acidB, rhein, aloeemodin, chrysophanol, physcionwere. Conclusions The UPLC-MS chromatographic method has the specialty of stability and repeatability, and it is suitable for analysis of Xiefei-Pingchuanling extract.
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Objective: To identify Euryales Semen and its closely related species using the ITS2 barcode. Method:The total genomic DNAs were extracted from twenty samples of Euryales Semen and its closely related species. The ITS2 regions of the samples were amplified and bidirectional sequenced. Obtained sequences were submitted to the GenBank with Sequin 12.3. ITS2 sequences of 102 samples belonging to thirty species were downloaded from GenBank. The 122 ITS2 sequences were aligned and the genetic distances were analyzed with MEGA 5.1. Identification analyses were performed using BLAST1 and nearest distance methods, and were presented intuitively by constructing neighbor-joining (NJ) tree. Result: The length of ITS2 region of Euryales Semen was 214 bp, which was only one haplotype. There was significant divergence of the ITS2 regions among the samples. The NJ tree showed that Euryales Semen could be obviously differed from its closely related species, which had good 408 monophyly. Conclusion: ITS2 regions as a DNA barcode can stably and accurately distinguish Euryales Semen from its closely related species and also provide a new technique to ensure clinical safety in utilization of tradi-tional Chinese medicines. The new exploration could broaden the application of DNA barcoding technology in identification of Traditional Chinese Medicine.
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<p><b>OBJECTIVE</b>To study the pollen morphological characteristics, viability test and storage character of the endangered plant Atractylodes lancea.</p><p><b>METHOD</b>Pollen grains morphologies of A. lancea were observed by scanning electron microscope. The optimum culture medium and viability determination methods were screened out by liquid culture and dyeing methods, and then the pollen germination capacities in different storage conditions were detected.</p><p><b>RESULT</b>The pollen grains are quasi-spherical, with tricolpate and spinous sculpture. The optimal culture medium was ME3 + 16% PEG4000 + 10% sucrose, in which the pollen germination capacity reached to 62.1%, while the other three dyeing methods were not able to be applied to detecting the pollen viability of A. lancea. The low storage temperature could significantly prolong the storage time of pollen of A. lancea. At -80 degrees C, pollen viability could be maintained for 60 days.</p><p><b>CONCLUSION</b>Liquid culture method is suitable for the determination of pollen germination of A. lancea, and the rate of pollen germination is closely related to the storage time and temperature. At last, this study provides a foundation for the artificial pollination and cultivating in wildness of A. lancea.</p>
Subject(s)
Atractylodes , Physiology , Endangered Species , Germination , Microscopy, Electron, Scanning , Plants, Medicinal , Physiology , Pollen , Physiology , Polyethylene Glycols , PharmacologyABSTRACT
Triterpenes, which have large application potential in the treatment of cancer, are the main active components of genuine medicinal material Alisma orientale (Sam.) Juzep. Farnesyl pyrophosphate synthase (FPPS) is one of the important rate-limiting enzymes in the synthetic pathway of triterpenes. In this study the FPPS full length cDNA of the A. orientale, was cloned via homology-based cloning approach and rapid amplification of cDNA ends (RACE). The full length of the FPPS cDNA was 1 531 bp (accession no. HQ724508), which contained a full 1 032 bp ORF that encoded 343 amino acids. The deduced protein sequence exhibited five conserved motifs, two of which is riched of Asp (DDXXD). The result of real-time quantitative PCR (QRT-PCR) showed that FPPS gene was expressed in different organs of A. orientale. The expression increased from October to the first ten-day period of December, and then decreased. The FPPS gene expression was higher in leaves but lower in leafstalk, tuber and root. HPLC analysis of active components 23-acetyl-alismol B of A. orientale. during different periods indicated that its change trend should be consistent with FPPS gene expression. It can be primarily deduced that FPPS gene should be an important control point in the synthetic pathway of Alisma terpenes. This study may facilitate the quality of medicinal plants through gene engineering in the future.