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The aim of this study was to evaluate the seed quality of C.deserticola and establish quality grading rules of seeds by detecting the impacts of different processing methods on the contents of the effective components of C.deserticola for optimizing the suitable processing method.The seed quality was judged by thousand-kernel weight,empty embryo rate and water content.The samples were preliminary processed by freeze-drying,natural drying and hot air circulation drying respectively,and the content of phenylethanoid glycosides was determined by HPLC-UV.The seed quality classification standard of C.deserticola was established,and the seeds were divided into three grades based on the standard.It was found that freeze-drying method was optimum,featuring less effective component loss,beautiful appearance of herbal piece,crisp texture and fast drying.In conclusion,this study laid a foundation for the quality control of the seeds of C.deserticola with the provision of scientific evidence for the initial processing of the fresh product.
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In this article, the method of quick and accurate identification ofAstragalus membranaceus (Fisch.) Bge. var.mongholicus (Bge.) Hsiao in different growth years by electronic nose (e-nose) technique and chemical analysis was developed. An e-nose was used to detect the odors ofA. membranaceus. var.mongholicus samples in two-growth-year, seven-growth year and more than a decade growth years for establishing the classification model of response characteristics. Principal component analysis (PCA) and discriminant factor analysis (DFA) were performed to differentiate theA. membranaceus. var.mongholicus samples in different growth years. PCA was performed to investigate the difference in the chemical composition and quality of different growth years inA. membranaceus. var.mongholicus samples. The results of PCA and DFA analyses for e-nose demonstrated that the samples ofA. membranaceus. var.mongholicus in different growth years could be distinguished obviously. The contents of chemical composition were similar in same growth years inA. membranaceus. var.mongholicus and different from different growth years. The results of chemical composition analysis indicated that the identification forA. membranaceus. var.mongholicus in different growth years was significant for the quality control. E-nose technique could identify the samples ofA. membranaceus. var.mongholicus in different growth years rapidly, sensitively and intactly, and could be applied for the quality control of traditional Chinese medicine.
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This study was aimed to investigate the relationship between the quality ofRheum palmatum L. and ecological factors, in order to provide the basis for nurture and cultivation ofR. palmatumL. The ultra performance liquid chromatography (UPLC) was used in the determination of 8 effective components in samples from 3 majorR. palmatumL. producing provinces, which wereGansu,SichuanandQinghai province. The SPSS software was applied to analyze the effective components by the one-way analysis of variance. And the correlation analysis between the content and ecological factors was also conducted. The results showed that the pressure, relative humidity and temperature had close relationship with the content of bioactive compounds inR. palmatumL. The content of effective components inR. palmatumL. was positively related to pressure, relative humidity, and temperature. It was concluded that the quality ofR. palmatumL. in Gansu province was better. And the ecological factors affected the accumulation of effective components. This research provided experimental basis for the quality ofR. palmatumL. in different regions and ecological adaptation. The results demonstrated critical meaning forR. palmatumL. quality improvement, appropriate ecological division, and industrialized development promotion ofR. palmatumL.
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Objective To determine and compare the contents of astragaloside Ⅳ in different medicinal parts of Astragalus membranaceus var. mongholicus.Methods The contents of astragaloside Ⅳ in fibrous roots, root heads, taproots and whole roots of A.membranaceus var.mongholicus were determined by HPLC.Results The order of contents of astragaloside Ⅳ in A.membranaceus var.mongholicus was fibrous roots >whole roots >taproots >root heads;The content of astragaloside Ⅳ in A.membranaceus var.mongholicus from Inner Mongolia was 1.85 to 2.7 times of the standard which prescribed in 2010 edition of Chinese Pharmacopoeia.Conclusion Fibrous roots, which were discarded in traditional processing methods, can be used as raw material to extract astragaloside Ⅳ.This study may provide a reference for the harvest and produce of A.membranaceus var.mongholicus.
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This study provides the candidate sequences in the identification of Radix et Rhizoma Clematidis and its adulterants using DNA barcoding. We amplified and sequenced the region psbA-trnH, with the data of 284 sequences from GenBank, the differential intra- and inter-specific divergences, genetic distance, barcoding gap were used to evaluate five barcodes, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The results showed that psbA-trnH barcodes performed high identification efficiency and inter-specific divergences among the five different DNA barcodes. Analysis of the barcoding gap and NJ tree showed psbA-trnH was superior to other barcodes. Based on the identification and PCR amplification efficiency, psbA-trnH can be the ideal barcode to identify Radix et Rhizoma Clematidis and its adulterants accurately.
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To establish a new method for identifying genus of Lilium by DNA barcoding technology, ITS, ITS2, psbA-trnH, matK and rbcL sequences were analyzed in term of variation of inter- and intra-species, barcoding gap, neighbor-joining tree to distinguish genus of Lilium based on 978 sequences from experimental and GenBank database, and identification efficiency was evaluated by Nearest distance and BLAST1 methods. The results showed that DNA barcoding could identify different species in genus of Lilium. ITS sequence performed higher identification efficiency, and had significant difference between intra- and inter-species. And NJ tree could also divide species into different clades. Results indicate that DNA barcoding can identify genus of Lilium accurately. ITS sequence can be the optimal barcode to identify species of Lilium.
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Quality variation and ecotype classification of Chinese herbal medicine are important scientific problems in Daodi herbal medicine research. The diversity of natural environmental conditions has led to form unique multi-Daodi, multi-product areas that produce particular Chinese herbal medicine. China is one of three big American ginseng (Panax quinquefolium L.) producing areas worldwide, with over 300 years of application and 40 years of cultivation history. Long-term production practice has led to the formation of three big advocate produce areas in China: Northeast province, Beijing and Shandong. P. quinquefolium L. grown under certain environmental conditions will develop long-term adaptations that will lead to more stable strains (different ecotypes). P. quinquefolium L., can vary greatly in quality; however, the ecological mechanisms causing this variation are still unclear. Root samples were collected from four-year-old cultivated P. quinquefolium L. plants in the three major genuine (Daodi) American ginseng-producing areas of Northeast province, Beijing and Shandong province, China. Ultra-performance liquid chromatography was used to analyze the contents of eight ginsenosides (Rg1, Re, Rb1, Rb2, Rb3, Rc, Rd, Rg2). Data for nine ecological factors, including temperature, moisture and sunlight, were obtained from the ecological database of Geographic Information System for Traditional Chinese Medicine. Soil samples from the sampling sites were collected. Effective boron and iron, available nitrogen and potassium, as well as other trace elements and soil nutrients, were determined by conventional soil physicochemical property assay methods. Analytical methods of biostatistics and numerical taxonomy were used to divide ecotypes of the three main Panax quinquefolium L. producing areas in China based on ginsenoside content, climate, soil and other ecological factors. To our knowledge, this is the first time that ecological division of P. quinquefolium L. producing areas in China has ever been conducted. The results show that there are two chemoecotypes of P. quinquefolium L. in China: ginsenoside Rb1-Re from outside Shanhaiguan, and ginsenoside Rg2-Rd from inside Shanhaiguan. Similarly, there are two types of climatic characteristics: inside Shanhaiguan (Beijing, Shandong) and outside Shanhaiguan (Northeast). This suggests that the formation and differentiation of chemoecotypes of P. quinquefolium L. is closely related to variability of the climatic and geographical environment. Additionally, ecological variation of the three main producing areas, characteristics of two climatic ecotypes, and soil characteristics are also discussed and summarized. These results provide experimental scientific evidence of the quality variation and ecological adaptation of P. quinquefolium L. from different producing areas. They also deepen our understanding of the biological nature of Daodi P. quinquefolium L. formation, and offer novel research models for other multi-origin, multi-Daodi Chinese herbal medicines ecotypes. In addition, the results demonstrate the critical need for improving quality, appropriate ecological regionalization and promoting industrialized development of P. quinquefolium L.
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Synthetic biology of traditional Chinese medicine (TCM) is a new and developing subject based on the research of secondary metabolite biosynthesis for nature products. The early development of synthetic biology focused on the screening and modification of parts or devices, and establishment of standardized device libraries. Panax notoginseng (Burk.) F.H.Chen is one of the most famous medicinal plants in Panax species. Triterpene saponins have important pharmacological activities in P. notoginseng. Squalene epoxidase (SE) has been considered as a key rate-limiting enzyme in biosynthetic pathways of triterpene saponins and phytosterols. SE acts as one of necessary devices for biosynthesis of triterpene saponins and phytosterols in vitro via synthetic biology approach. Here we cloned two genes encoding squalene epoxidase (PnSE1 and PnSE2) and analyzed the predict amino acid sequences by bioinformatic analysis. Further, we detected the gene expression profiling in different organs and the expression level of SEs in leaves elicited by methyl jasmonate (MeJA) treatment in 4-year-old P notoginseng using real-time quantitative PCR (real-time PCR). The study will provide a foundation for discovery and modification of devices in previous research by TCM synthetic biology. PnSE1 and PnSE2 encoded predicted proteins of 537 and 545 amino acids, respectively. Two amino acid sequences predicted from PnSEs shared strong similarity (79%), but were highly divergent in N-terminal regions (the first 70 amino acids). The genes expression profiling detected by real-time PCR, PnSE1 mRNA abundantly accumulated in all organs, especially in flower. PnSE2 was only weakly expressed and preferentially in flower. MeJA treatment enhanced the accumulation of PnSEI mRNA expression level in leaves, while there is no obvious enhancement of PnSE2 in same condition. Results indicated that the gene expressions of PnSE1 and PnSE2 were differently transcribed in four organs, and two PnSEs differently responded to MeJA stimuli. It was strongly suggested that PnSEs play different roles in secondary metabolite biosynthesis in P. notoginseng. PnSE1 might be involved in triterpenoid biosynthesis and PnSE2 might be involved in phytosterol biosynthesis.
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DNA barcoding is a rapidly developing frontier technology in the world and will be useful in promoting the quality control and standardization of traditional Chinese medicine. Until now, many studies concerning DNA barcoding have focused on leaf samples but rarely on Chinese herbal medicine. There are three issues involved in DNA barcoding for traditional Chinese medicinal materials: (1) the extraction methods for total DNA of the rhizomes of the medicinal materials; (2) intra-specific variation among samples from different places of origin; (3) accuracy and stability of this method. In this study, Gentianae Macrophyllae Radix was used to verify the stability and accuracy of DNA barcoding technology. Five regions (ITS2, psbA-trnH, matK, rbcL, and ITS) were tested for their ability to identify 86 samples of Gentianae Macrophyllae Radix and their adulterants. After improving the DNA extraction method, genomic DNA from all samples was successfully obtained. To evaluate each barcode's utility for species authentication, PCR amplification efficiency, genetic divergence, and species authentication were assessed. Among all tested regions only ITS2 locus showed 100% of PCR amplification and identification efficiencies. Based on the established method, we successfully identified two samples of Gentianae Macrophyllae Radix bought in pharmacy to the original species.
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In this review, the authors summarized the new technologies and methods for identifying traditional Chinese medicinal materials, including molecular identification, chemical identification, morphological identification, microscopic identification and identification based on biological effects. The authors introduced the principle, characteristics, application and prospect on each new technology or method and compared their advantages and disadvantages. In general, new methods make the result more objective and accurate. DNA barcoding technique and spectroscopy identification have their owner obvious strongpoint in universality and digitalization. In the near future, the two techniques are promising to be the main trend for identifying traditional Chinese medicinal materials. The identification techniques based on microscopy, liquid chromatography, PCR, biological effects and DNA chip will be indispensable supplements. However, the bionic identification technology is just placed in the developing stage at present.
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Código de Barras de DNA Taxonômico , Medicamentos de Ervas Chinesas , Química , Medicina Tradicional Chinesa , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da PolimeraseRESUMO
<p><b>OBJECTIVE</b>To clone and sequence the open reading frame of cycloartenol synthase (CAS) from Huperzia carinata.</p><p><b>METHOD</b>After searching the transcriptome dataset of H. carinata, one unique sequence containing oxide squalene cyclases domain was discovered. The primers were designed according to the cDNA sequence of CAS from the dataset. And then, the open reading frame of CAS was cloned by RT-PCR strategy with the template of mixed RNA extracted from root, stem and leaf of H. carinata. The bioinformatic analysis of this gene and its corresponding protein was performed.</p><p><b>RESULT</b>One unique sequence of CAS, named as HcCAS1 (GenBank accession number JN790125) , was cloned from H. carinata. The open reading frame of HcCAS1 consists of 2 474 bp, encoding one polypeptide with 757 amino acids.</p><p><b>CONCLUSION</b>This study cloned and analyzed CAS from H. carinata for the first time. The result will provide a foundation for exploring the mechanism of sterol biosynthesis in Huperziaceae plants.</p>
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Sequência de Aminoácidos , Clonagem Molecular , Biologia Computacional , Evolução Molecular , Huperzia , Genética , Transferases Intramoleculares , Química , Genética , Metabolismo , Modelos Moleculares , Dados de Sequência Molecular , Filogenia , Estrutura Secundária de ProteínaRESUMO
<p><b>OBJECTIVE</b>To establish a HPLC fingerprint of water-soluble constituents of Fritillaria unibracteata.</p><p><b>METHOD</b>Zorbax SB Aq C18 chromatographic column (4. 6 mm x 250 mm, 5 microm) was adopted for gradient elute with the mobile phase consisting of methanol and water. The flow rate was 1.0 mL x min(-1); the detection wavelength was 260 nm, and the temperature of sample manager was set at 25 degrees C. Similarity Evaluation System for Chromatographic Fingerprint of traditional Chinese medicine (version 2.0) published by the State Pharmacopeia Committee of China was adopted for the fingerprint analysis on the 11 batches of F. unibracteata herbs.</p><p><b>RESULT</b>The 11 batches of F. unibracteata herbs had 14 common peaks, nine of which were identified with good separating degrees. The similarities of the 11 batches were more than 0. 970, with good quality homogeneity.</p><p><b>CONCLUSION</b>The method is so accurate, highly reproducible and stable that it is suitable for the comprehensive quality evaluation of F. unibracteata herbs.</p>
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Cromatografia Líquida de Alta Pressão , Métodos , Dermatoglifia , Medicamentos de Ervas Chinesas , Química , Fritillaria , Química , Medicina Tradicional Chinesa , Solubilidade , Água , QuímicaRESUMO
<p><b>OBJECTIVE</b>To develop a high performance liquid chromatography coupled with a photodiode array detector (HPLC-DAD) method for simultaneous determination of 7 nucleosides and nucleobases in Fritillaria taipaiensis.</p><p><b>METHOD</b>The analyses were performed on an Agilent Zorbax-SB-Aq-C18 column (4.6 mm x 250 mm, 5 microm) eluted with water and methanol in gradient mode. The flow rate was 1.0 mL x min(-1). The detection wavelength was 260 nm. The temperature of sample manager was set at 25 degrees C, and the injection volume was 20 microL.</p><p><b>RESULT</b>The investigated compounds including uracil, cytidine, uridine, guanosine, thymidine, adenosine and adenine were shown good linearity (r > or = 0.999 8) over the tested ranges. The average recoveries were within 96.96% - 103.5% with RSD < or = 3.8%.</p><p><b>CONCLUSION</b>The accuracy, stability, repeatability and average recovery of the method are satisfying, and the seven nucleosides and nucleobases components in F. taipaiensis can be rapidly and accurately quantified by HPLC-DAD. This work provided helpful information for comprehensive quality evaluation of F. taipaiensis.</p>
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Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas , Química , Fritillaria , Química , Nucleosídeos , Reprodutibilidade dos Testes , Água , QuímicaRESUMO
The paper reports the establishment of a method for simultaneous determination of peimisine and sipeimine contents in Fritillaria walujewii Regel and Fritillaria pallidiflora Schrenk. The analyses were performed on an ultra-performance liquid chromatography with evaporative light scattering detection (UPLC-ELSD), equipped with a binary solvent manager, a sampler manager and a column compartment, and connected to Waters Empower 2 software. An Acquity UPLC BEH C18 column (100 mm x 2.1 mm, 1.7 microm) was used for all analysis. The investigated compounds were separated with gradient mobile phase consisting of acetonitrile-0.02% triethylamine-water. The temperature of sample manager was set at 25 degrees C. Drift tube temperature was 40 degrees C, and spray parameter was 40% with injection volume of 1 microL. The investigated compounds including peimisine and sipeimine had good linearity (r > or = 0.9991) over the tested ranges. The average recovery was 94.5% and 98.1% with RSD < or = 2.36%. The UPLC-ELSD method is simple, sensitive and accurate with good repeatability, which is available for quality control of F. walujewii Regel and F. pallidiflora Schrenk.
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Objective To observe the effects of modified Yigong Powder (MYP) combined with chemotherapy on transplanted hepatocarcinoma of mice and to study its mechanisms. Methods Hepatocarcinoma 22 mouse model was established and then was used to observe the attenuating and synergic effects of MYP when applied together with 5-fluorouracil (5-FU). After 8 days of treatment,the tumor-inhibiting rate,activity of natural killer (NK)cells and interleukin-2 (IL-2) and small intestinal malondialdehyde (MDA) content were examined. Results MYP combined with 5-FU could increased the tumor-inhibiting rate to some extent and improve the immune function by increasing immune organ weight and increasing the activity of NK cells and IL-2. MYP combined with 5-FU could also reduce the 5-FU-induced intestinal injuries by relieving the damage of free radicals and inhibiting the lipid peroxidation and a good prognosis was expected in tumor-bearing animals treated with chemotherapy. Conclusion MYP exerts an attenuating and synergic effect when used together with 5-FU in treating tumor-bearing mice and its mechamism may be related to the improvement of immune function and reduction of intestinal injuries.