Chemical profiling of Coptis rootlet and screening of its bioactive compounds in inhibiting Staphylococcus aureus by UPLC-Q-TOF/MS.

Coptis chinensis Franch. is a traditional Chinese medical herb, only the rhizome is used in clinical. However, the other discarded parts, such as rootlet, still contain a large amount of active ingredients. In this article, UPLC-ESI-Q-TOF/MS method was used to identify components of Coptis rootlet, the differences in the contents of four major alkaloids in Coptis rootlet and Coptis rhizome were compared using HPLC. Then the antibacterial effect of the ethanolic extracts of Coptis rootlet and Coptis rhizome on Staphylococcus aureus (S. aureus) were investigated. Finally, S. aureus were cultured for a period of time in the environment of minimum inhibitory concentration (MIC) of Coptis rootlet, extracting intracellular substances, screening for effective antibacterial ingredients. By comparison with secondary mass spectrogram of reference substance, a total of 29 components were identified in Coptis rootlet, the contents of berberine, palmatine, coptisine, epiberberine in rootlet and rhizome were determined and compared. 6 alkaloids were found in the cells of S. aureus, which is the main antibacterial compounds of Coptis rootlet. This study proved the utility of the Coptis rootlet, which is beneficial to the more comprehensive utilization of medicinal plant resources.

[Research for dynamic changes of growth and alkaloids of Coptis chinensis].

With Coptis chinensis in high-yielding soil as the object,the growth regularity of plant and dynamic change of alkaloid content was studied. The plant growth model of C. chinensis was constructed. The plant height equation was y=3.030 9+0.732 6x-0.009 6x²,the number of leaves equation was y=111.882 6-2 234.881 7/x+15 218.960 8/x²-31 740.960 8/x³,the leaf area equation was y=-217.136 1+30.552 2x-0.359 0x²,the roots talk biomass equation was y=-2.748 8+0.210 3x+0.006 4x²,the number of rootstalk equation was y=-1.246 0+0.192 6x+0.000 8x²,the fibrous root biomass equation was y=-4.973 5+0.589 4x -0.002 6x². The results indicated that the number of leaves and leaf area were increasing continuously after seedling transplanting,the leaf area of 3-year-old C. chinensis reached a maximum value of 425.83 cm²/plant,after declining.The number of leave of 5-year-old C. chinensis reached a maximum value of 70.91. With the increasing of years of growth, the number of rootstalk and rootstalk biomass of C. chinensis was increasing continuously. The biomass growth of 3-year-old and 4-year-old rootstalk was the fastest in the whole development stage of C. chinensis,the annual increase of more than 300%. The change curve of rootstalk number, rootstalk biomass and fibrous root biomass in the whole growth stage was a s-type.The dry matter partition of leafwas the highest in 1-year-old C. chinensis, and then gradually decreased,the change trend of dry matter partition of rootstalk was just the opposite, the dry matter partition of fibrous root increases with the increase of the growing year, reaching the maximum value in 3-year-old, then gradually lower trend. The root-shootratio of 1-year-old C. chinensis was the smallest, then gradually increases, the growth center gradually shifted to the roots from stems and leaves, The weight of underground part of 3-year-old C. chinensis exceeded the aboveground part, the 5-year-old C. chinensis root-shoot ratio reached the maximum value of 1.91:1.With the increasing of years of growth, the contents of coptisine, berberine, epiberberine and palmatine in rootstalk was increasing continuously. The jatrorrhizine content in 2-year-old C. chinensis was significantly lower than that in other years, the content was no significant change after that. The columbamine content reached a maximum value in 3-year-old C. chinensis,then the decreased gradually. The content of magnoflorine gradually increased and reached maximum value in 5-year-old C. chinensis.

[Cultural regionalization for Coptis chinensis based on 3S technology platform â . Study on growth suitability for Coptis chinensis based on ecological factors analysis by Maxent and ArcGIS model].

At the urgent request of Coptis chinensis planting,growth suitability as assessment indicators for C. chinensis cultivation was proposed and analyzed in this paper , based on chemical quality determination and ecological fators analysis by Maxent and ArcGIS model. Its potential distribution areas at differernt suitability grade and regionalization map were formulated based on statistical theory and growth suitability theory. The results showed that the most suitable habitats is some parts of Chongqing and Hubei province, such as Shizhu, Lichuan, Wulong, Wuxi, Enshi. There are seven ecological factor is the main ecological factors affect the growth of Coptidis Rhizoma, including altitude, precipitation in February and September and the rise of precipitation and altitude is conducive to the accumulation of total alkaloid content in C. chinensis. Therefore, The results of the study not only illustrates the most suitable for the surroundings of Coptidis Rhizoma, also helpful to further research and practice of cultivation regionalization, wild resource monitoring and large-scale cultivation of traditional Chinese medicine plants.

[Bacteria community in different aged Coptis chinensis planting soil revealed by PCR-DGGE analysis].

In order to reveal the cause of disease occurred in the process of Coptis chinensis growth, this paper studied the bacterial species diversity index of different aged rhizospheric and non-rhizospheric soil planting normal or sick C. chinensis by using PCR-DGGE technique. The representative DGGE bands were chosen to be cloned, and sequenced, the phylogeny were constructed. The results showed that the bacterial communities were very different between the normal and diseased soil samples of C. chinensis, and the diversity index (H) of diseased soil samples were higher than that of normal soil samples. Sequencing analysis of representative cloned DGGE bands showed that the unculturable bacteria were the dominant groups, and bacteria belonged to genus Bacillus, Acidovorax, Acinetobacter, uncultured Kluyvera, and uncultured Comamonas were also existing, but the reported plant pathogenic bacteria were not found in the C. chinensis planting soil. The density and brightness of clone band d in diseased soil samples was higher than that in normal soil sample, and sequencing analysis showed that it belonged to genus Acidovorax. Obviously, during the process of C. chinensis growth, the rhizospheric bacteria population changed, and the quantity of bacteria belong Acidovorax increased, which probably resulted in the disease occurred during C. chinensis growth.

[Classification study of Coptis chinensis based on quantitative physical property characteristics of appearance and internal quality evaluation].

OBJECTIVE: Combining the quantitative physical property characteristics of the appearance with the internal quality evaluation index, its aims to provide experimental basis for the classification and quality evaluation of Coptis chinensis. METHOD: Fourteen batches of C. chinensis from different areas were respectively measured in size (total length, total width, root length, taproot diameter, branch number, branch length, branch diameter, length of the bridge, weight), color (external color, internal color), content (epiberberine, coptisine, palmatine, berberine). Then the determination data were evaluated by spss principal component analysis and cluster analysis. RESULT: Three principal components were extracted from the original data. The principal component analysis results showed that the characteristic elements might be the total length, main root length, taproot diameter, branch length, weight, the total color value of the appearance and content of epiberberine and berberine. The results of cluster analysis showed that 14 batches of samples could be clustered reasonably into two groups. In terms of the appearance and quality, there were some differences between in the geo-authentic and non-authentic producing areas of C. chinensis. CONCLUSION: The method which was combining the quantitative physical property characteristics of the appearance with the internal quality evaluation index, and through the processing of mathematical statistics, could be used for the the classification of C. chinensis.

[Huanglian places of origin distribution and change from herbal-works].

The herbal-works recorded that the origin production places of Huanglian had gone through dynamic condition of distribution and change. During Pre-Qin and Qin-Han Dynasty, the origin production places of Huanglian were small and limited. During Weijin-Nanbei Dynasty, the origin production places of Huanglian grew in number gradually. During Sui-Tang and Song-Yuan Dynasty, the origin production places of Huanglian were enlarged quickly. During Ming and Qing Dynasty, the origin production places of Huanglian were expended small-size. The origin production places of Huanglian were enlarged because its functions and demands added and planting-technology skills developed. The records of the herbal-works were not complete, we should pay attention to the other works, so as to ascertain the realorigin production places of Huanglian exactly and scientifically.

[Investigation and protection for endangered Coptis deltoidea].

OBJECTIVE: To investigate the history of the medicinal uses, resources, distribution, habitat and population characteristic of Coptis deltoidea, and provide basis for the protection and rational development and utilization of Radix Coptidis Deltoideae. METHOD: The relevant literature and data was scrutinized and herbarium was compared, interview and field survey methods were carried out. RESULT: The medicinal history, resources, distribution, population characteristic and protective strategy of C. deltoidea were summarized. The sustainable development of C. deltoidea was discussed. CONCLUSION: The resource is endangered, the germplasm resources should be intentionally protected and ensure the sustainable development and utilization of C. deltoidea.

[Breeding system, morphological and anatomical characteristics of Coptis deltoidea].

OBJECTIVE: To study the morphological and anatomical characteristics of Coptis deltoidea based on the research of its breeding system. METHOD: The flowering process of C. deltoidea was observed dynamically and the out-crossing index was estimated, using Motic BA200 microscope to observe the morphologic characteristics of the mixed bud, stolon and pollen was observed microscopically, and compared with those of C. chinensis. RESULT: The results showed that the breeding system of C. deltoidea was facultative hybridization, it could bloom normally and fruit while cannot form seeds. And there were no statistical differences in the number of stamens, pistils, ovules, pollens in a stamen, pollens in a flower, pollen-ovule ratio, and the pollen grain size between C. deltoidea and C. chinensis. However the pollen of C. deltoidea developed anomaly, most of the pollen grains in it were surface depression, the texture was unsharp or broken, the pollen could not germinate. The lateral bud on the lower side of the mixed bud formed in the bud stage. Pericyclic fibers in the stolon (the vegetative propagation branch) shaped like a cap, and all the shaped-caps nearly formed a ring. CONCLUSION: The abnormal development of the pollen could be the mainly cause to the no formation of seeds in C. deltoidea. The lateral bud forming in the bud stage then developing the stolon is the characteristics of the asexual propagation. Pericycle fibers in the stolon nearly forming a ring is a secondary character to accommodate the vegetative propagation of C. deltoidea.

[Study on the autointoxication of aquatic extracts from different parts of Coptis chinensis].

OBJECTIVE: To study the autointoxication of aquatic extracts from different parts of Coptis chinensis. METHODS: By measuring the effects on seed germination, seedling growth and adult-plants' growth, the autointoxications of root exudates, rhizome extracts, plant residue decomposition products and aqueous extracts from rhizosphere soil were studied. RESULTS: The aquatic extracts from different parts of C. chinensis could inhibit the nutritional growth stage of C. chinensis. CONCLUSIONS: Autointoxication is one of the factors leading to continuous cropping obstacle, and this study is instructive to the management and application of C. chinensis in agricultural productivity.

[Effects of slow release-fertilizers on yield, nutrient contents and quality of Coptis chinensis].

OBJECTIVE: To study the effects of four kind of slow-release fertilizers on yield and quality of Coptis chinensis. METHOD: One to three years C. chinensis was fertilized with slow-release fertilizers twice in April and in September. The yield and nutrient content along with quality of C. chinensis were measured after two years growth. RESULT AND CONCLUSION: All of the slow-release fertilizers increased the yield obviously, and the effect of SRF1 and SRF4 is the most significant. Comparing with control group, the N content in aerial part of 1-2 year-old C. chinensis treated with SRF1 and SRF4 was lower and P and K were higher than that of control group, and the N content in aerial part of 3 year-old C. chinensis treated with SRF1 and SRF4 was higher and P and K were higher than that of control group; The N content in the root of land 3 year-old C. chinensis treated with SRF1 and SRF4 showed no significant difference comparing with control group, and P and K were lower than that of control group, the N and P content in root of 2 year-old C. chinensis treated with SRF1 and SRF4 was higher and K were lower than that of control group. After two years growth berberine content of C. chinensis treated with SRF1, SRF2 and SRF3 were significantly increased, and total alkaloid content of C. chinensis treated with SRF1, SRF3 and SRF4 were significantly increased. We recommend that SRF4 is used as the special fertilizer for 1-year-old C. chinensis, and the SRF1 and SRF4 for 2-year-old C. chinensis, and the SRF1 for 3-year-old C. chinensis.

Coptis teeta-based agroforestry system and its conservation potential: a case study from northwest Yunnan.

Coptis teeta (Ranunculaceae), is a nontimber forest product (NTFP) that only grows in northwest Yunnan and northeast India. Its tenuous rhizome, known as "Yunnan goldthread" in the traditional Chinese medicine system, has been used as an antibacterial and as an antiinflammatory medicine for a long time. The increasing demand has resulted in commercial harvesting pressure on wild populations that were already dwindling as a result of deforestation, and wild populations are at risk of extinction. Fortunately, there exists at least 2000 hectares of a C. teeta-based agroforestry system initiated by the Lisu people in Nujiang, northwest Yunnan. This cultivation supplies us with a valuable study case for the balance between conservation and sustainable use. This case study investigated the traditional management system and history of C. teeta in Nujiang through ethnobotanical methods and field investigation. We also contrasted initial costs, economic returns, and labor demands for C. teeta cultivation with other major land uses in the region. Compared with swidden agriculture, the major land-use type in the region, C. teeta cultivation offers high economic returns and low labor and initial costs; moreover, C. teeta cultivation does not interfere with subsistence agricultural duties. This agroforestry system reflected that the cultivation of NTFPs is a conservation strategy for maintaining forest diversity, while providing a stable economic return to local forest communities, and indicates how local people manage biodiversity effectively.

[Quality assessment for Coptis chinensis with different cultivating system].

OBJECTIVE: To evaluate the quality of Coptis chinensis planted with four kinds of cultivating system. METHOD: The samples, which were planted below the artificial shading, the forest of Chinese ceder wood, Mmulbery wood, Magnolia officinal wood were chosen respectivley, UV and HPLC were used to determine of total alkaloids and berberine of samples. RESULT: There was no significant difference among four kinds of cultivating system on the contents of total alkaloids and berberine. The contents of total alkaloids and berberine were obviously higher than in the criterion of Chinese Pharmacopoeia. CONCLUSION: The roots of C. chinensis planted in the four kinds of planting system can be used as medicinal material of high quality.

[Quality analysis and evaluation of Rhizoma Coptidis under different cultivation conditions].

AIM: To develop methods for the fingerprint analysis of Rhizoma Coptidis and the determination of berberine, palmatine and jatrorrhizine in Rhizoma Coptidis, and analyze the contents of these three alkaloids in Rhizoma Coptidis under different cultivation conditions, from different areas and processed with different methods. METHODS: Two methods (HPLC-UV and HPLC-MS) have been developed and used in fingerprint analysis of Rhizoma Coptidis. An HPLC method was used to determine the contents of three alkaloids. RESULTS: With HPLC-MS techniques, seven major chromatographic peaks in the fingerprint analysis of Rhizoma Coptidis were identified by their MS spectra and compared with the reference standards. In different cultivation conditions, shading conditions and growing ages have obvious influence on the contents of three alkaloids in Rhizoma Coptidis, while planting density was not the major factor that influenced the contents of three alkaloids. The contents of three alkaloids of Coptidis samples were almost higher than those of Coptidis reference material. For Coptidis samples from different cultivation area, the contents of these three alkaloids were different greatly. For Coptidis samples processed with different methods, the contents of three alkaloids were not influenced obviously by processing methods. CONCLUSION: The results showed that the ecology cultivation method to replace the traditional shading method was feasible and provided the theoretical foundation for scientifically processing Rhizoma Coptidis.

[Analysis and characterization of fingerprint infrared spectra for ecology-planted Coptis chinensis].

The fingerprint features of infrared spectra for Coptis chinensis in different shading conditions (shelf, firry wood, corn wood) and with different ages (one year to five years) were studied. According to the ratio of characteristic peaks, the rules of the influence of shadings and ages on Coptis chinensis growth were illustrated, which confirms the feasibility of ecological planting technique for Coptis chinensis. Fourier transform infrared (FTIR) spectroscopy was used to analyze the influence of growing conditions on the growth of Coptis chinensis. The shadings and ages have no obvious influence on the quality of Coptis chinensis root, which is consistent with the results of HPLC. The quality of Coptis chinensis with ecological planting is identical with those in shelves. Infrared technique provides a meaningful guide to standardized planting of medicinal plants.

[Dynamic change of soluble saccharides and amino acids during the growing period of Coptis chinensis].

OBJECTIVE: To study the changes of soluble saccharides and amino acids during the growing period of Coptis chinensis. METHOD: Amino acid contents were estimated by Indene tri-ketone colorimetric method; saccharide contents were determined by anthracene ketone method. RESULT AND CONCLUSION: The contents of soluble saccharides and amino acids in different organs were low in plants of 1-2 years old. And they were increased in plants of 3-4 years old. At maturation (5 years), the contents of amino acids kept on increasing in rhizome and flower stalk, while decreased in fibre and leaves, and the contents of soluble saccharides showed the downtrend in rhizome, leaves and fibre, except the uptrend in flower stalk.

[Preliminary studies on the nutrient characters of different kinds of Chinese goldthread soil].

OBJECTIVE: To study the changes of nutrient contents in Chinese goldthread soil before and after planting Coptis chinensis. METHOD: The soil in Shizhu, Chongqing, was analyzed for nutrient contents. RESULT AND CONCLUSION: The results indicated that the preplanted soil was strongly acidic and the levels of organic matter, total nitrogen and potassium were high. However, the content of total phosphorus was very low; so were the contents of other eleven nutrients. Variations of nutrients except zinc and molybdenum were large. After four years of planting C. chinensis, the soil pH was increased. Contents of organic matters, manganese, zinc and boron were increased, while the contents of iron and copper were decreased. In general, the variance coefficient of all nutrient elements and the difference of soil fertility were decreased.

[Application and prospect on the researches of ecological cultivation of Coptis chinensis].

Traditional cultivation of Coptis chinensis was carried out under shield by disafforestation, which has been used for over 300 years and lead to the severe destruction of natural environment. Several ecological modes for cultivation of Coptis chinensis have been developed, which increase the yields of Coptis chinensis, protect the resources of forest, and obtain economic and ecologic benefit.

[Ecological technique of cultivating Coptis chinensis in woods and co-cultivating it with maize].

OBJECTIVE: To establish ecologically new modes of cultivating Coptis chinensis in woods and co-cultivating it with maize. METHODS: Based on the experience obtained from plot comparative test and production test, we used application-oriented research methods, and established new Coptis chinensis cultivation techniques that protect the natural environment. RESULTS: Coptis chinensis was harvested 6 years after cultivation. Yield of Coptis chinensis cultivated in forest (69.5 kg per 0.067 hm2) was higher with 3.7% than that in controls which cultivated under shed (67 kg). Yield of Coptis chinensis co-cultivated together with maize (168.4 kg per 0.067 hm2) was lower with 15.8% than that in controls which cultivated under shed (200 kg). CONCLUSION: The new cultivation modes of Coptis chinensis is an ecologically new technique that assures the good growth of medicinal plants, cereals, forest and animal husbandry.

[Quality assessment for Coptis chinensis planted with ecological techniques].

OBJECTIVE: To evaluate the quality difference between Coptis chinensis planted with ecological techniques and shelf planted Coptis chinensis. METHODS: Ultraviolet-visible spectrophotometry, alcohol extract mensuration, moisture mensuration, and ash mensuration were used to determine the contents of total alkaloids, alcohol extract, water, and total ash of Coptis chinensis, which were planted in shelf, Rhus chinensis wood, M mulbery wood, corn wood, Magnolia officinalis wood, fruiter wood, shading net, and firry wood, respectively. RESULTS: The contents of total alkaloids and alcohol extract of Coptis chinensis Table planted with ecological techniques were higher than those of Coptis chinensis planted in shelf. The contents of water and total ash were less than 12% and 5%, respectively, which met the provisions of the pharmacopoeia. CONCLUSION: The quality of Coptis chinensis planted with ecological techniques is similar to that of Coptis chinensis planted in shelf. These ecological techniques for Coptis chinensis have become mature and practical.

[Technique of storing Coptis chinensis seeds in damp sand under shed and fine cultivation of seedling].

OBJECTIVE: To develop technique of storing Coptis chinensis seeds in damp sand under shed and fine cultivation of seedling and to enhance the cultivation seedling rate from seeds of Coptis chinensis. METHODS: With the old technique of seedling in woods as control, we screened the new method for storing seeds and cultivating seedlings. RESULTS: Seeds were picked up at May and stored in damp sand under shed until November, and then planted with technique of fine cultivation of seedling. The germination rate of seeds was up to 98%. One hundred and forty thousands seedlings could be getting from per kilogram seeds. CONCLUSION: Compared to the old method of cultivating seedling in woods, the technique of fine cultivation of seedling significantly increased the rate of cultivation seedlings by 8 times. This technique has been widely applied.