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AIM To establish an UPLC-MS/MS method for simultaneous content determination of protocatechuic acid,epicatechin,chlorogenic acid,quercitrin,gaultherin and gaultheroside A in Gaultheria leucocarpa var.yunnanensis.METHODS The analysis was performed on a 40℃thermostatic Waters BEH C18 column(100 mm×2.1 mm,1.7 μm),with the mobile phase comprising of water(containing 0.1%formic acid)-acetonitrile(containing 0.1%formic acid)flowing at 0.3 mL/min in a gradient elution manner,and electron spray inoization source was adopted in positive and negative ion scanning with multiple reaction monitoring(MRM)mode.Hierarchical cluster analysis(HCA)and principal component analysis(PCA)was used to screen important components that affect the quality of medicinal materials.RESULTS Six constituents showed good linear relationships within their own ranges(R2≥0.998 2),whose average recoveries were 98.76%-101.88%with the RSDs of 1.0%-2.5%.The constituents of G.leucocarpa in the roots and aerial parts were quite different.Gaultherin,epicatechin and protocatechuic acid may be the quality mark constituents of G.leucocarpa.CONCLUSION This accurate and efficient method can be used for the quality control of G.leucocarpa.
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Objective The contents of 11 nucleosides and base components in 10 batches of samples from 5 provinces(cities)including Chongqing,Yunnan and Shaanxi were determined,and the differences in nucleosides and base components in Fritillaria taipaiensis were compared by chemometric analysis,and the quality was comprehensively evaluated,so as to provide a reference for the cultivation of excellent varieties and the selection of medicinal materials.Methods Nucleoside and base components were extracted from Fritillaria taipaiensis by ultrasonication in aqueous solutions,and the content of each component was determined by HPLC-DAD method.The origin was classified by principal component analysis(PCA)and hierarchical cluster analysis(HCA).Partial least squares discriminant analysis(PLS-DA)was used to determine the differentiated index components in Fritillaria taipaiensis.Then the differences in the contents of the index components among samples from different origins were compared.Results It was found that 11 nucleoside and base components differed significantly among different origins of Fritillaria taipaiensis.Principal component analysis and hierarchical cluster analysis indicated that all samples could be clustered into 4 categories.Five characteristic components,including uracil,cytosine,uridine,inosine,and adenosine,were identified by PLS-DA.The nucleosides and bases in samples from Chongqing and Hubei were relatively high,and the quality of the samples was comparatively superior.Conclusion This method is simple,reproducible,accurate and reliable.It has screened out the index nucleoside and base components in the identification of Fritillaria taipaiensis of different origins,which can be used to initially elucidate the differences of samples between different origins.Additionally,it can better reflect the quality of Fritillaria taipaiensis,and can provide reference for the selection of procurement origin and the quality control for Fritillaria taipaiensis.
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Objective To explore the classification and characteristics of common syndromes of primary liver cancer and provide reference for clinical treatment.Methods Collect the four diagnostic information of patients with primary liver cancer from two top three TCM hospitals in Henan Province,and build a database.Using Lantern 5.0 software,based on two-step hidden tree analysis,a hidden structure model was constructed,and common syndromes of primary liver cancer were extracted through comprehensive clustering.SPSS 23.0 software was used for factor analysis and systematic cluster analysis to infer the potential syndromes.Combined with the results of different methods and professional knowledge,the syndrome classification of primary liver cancer was determined.Results A total of 1353 patients with 105 symptoms of primary liver cancer were included.59 symptoms with an analysis frequency≥40 were included to construct a hidden structure model,24 hidden variables were obtained,and 5 common syndromes were obtained by comprehensive clustering,namely,qi deficiency syndrome,liver depression and qi stagnation syndrome,blood stasis syndrome,water dampness stagnation syndrome,liver and gallbladder damp heat syndrome.20 common factors were obtained by factor analysis for symptoms with frequency>3%,and 8 common syndromes were inferred by cluster analysis with common factors.7 common syndromes and characteristics were finally determined by combining different methods and expertise.Conclusion The common syndromes of primary liver cancer are qi deficiency syndrome,liver depression and qi stagnation syndrome,blood stasis syndrome,water dampness stagnation syndrome,yin deficiency syndrome,liver and gallbladder damp heat syndrome,spleen deficiency and dampness stagnation syndrome.The results objectively reflect the actual situation of patients with primary liver cancer,and can provide reference for the treatment of primary liver cancer based on syndrome differentiation.
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ObjectiveTo establish the identification method of Dalbergiae Odoriferae Lignum(DOL) and its counterfeits by nuclear magnetic resonance hydrogen spectrum(1H-NMR) combined with multivariate statistical analysis. Method1H-NMR spectra of DOL and its counterfeits were obtained by NMR, and the full composition information was established and transformed into a data matrix, and the detection conditions were as follows:taking dimethyl sulfoxide-d6(DMSO-d6) containing 0.03% tetramethylsilane(TMS) as the solvent, the constant temperature at 298 K(1 K=-272.15 ℃), pulse interval of 1.00 s, spectrum width of 12 019.23 Hz, the scanning number of 16 times, and the sampling time of 1.08 s. Similarity examination and hierarchical cluster analysis(HCA) were performed on the data matrix of DOL and its counterfeits, and orthogonal partial least squares-discriminant analysis(OPLS-DA) was used to analyze the data matrix and identify the differential components between them. In the established OPLS-DA category variable value model, the category variable value of DOL was set as 1, and the category variable value of the counterfeits was set as 0, and the threshold was set as ±0.3, in order to identify the commercially available DOL. The OPLS-DA score plot was used to determine the types of counterfeits in commercially available DOL, and it was verified by thin layer chromatography(TLC). ResultThe results of similarity analysis and HCA showed that there was a significant difference between DOL and its counterfeits. OPLS-DA found that the differential component between DOL and its counterfeits was trans-nerolidol. The established category variable value model could successfully identify the authenticity of the commercially available DOL. The results of the OPLS-DA score plot showed that there were heartwood of Dalbergia pinnata and D. cochinchinensis in the commercially available DOL, and were consistent with the TLC verification results. ConclusionThere is a phenomenon that heartwood of D. pinnata and D. cochinchinensis are sold as DOL in the market. 1H-NMR combined with multivariate statistical analysis can effectively distinguish DOL and its counterfeits, which can provide a reference for the identification of them.
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The present study established the ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method for simultaneous determination of the content of eight major active components in Caesalpinia decapetala and performed the quality evaluation of C. decapetala from different habitats with the chemical pattern recognition. The analysis was carried out on a Waters BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) at 40 ℃, with the mobile phase of water containing 0.1% formic acid(A) and acetonitrile containing 0.1% formic acid under gradient elution, the flow rate of 0.3 mL·min~(-1), and the injection volume of 1 μL. The electrospray ionization(ESI) source in the negative mode and multiple reaction monitoring(MRM) were used for MS quantitative analysis. The content results were analyzed by the hierarchical cluster analysis(HCA) and principal component analysis(PCA) for the evaluation of the quality difference. Eight components showed good linear relationships within their respective concentration ranges(r>0.999), with the average recoveries of 96.85%-103.4% and RSD of 0.52%-2.8%. The analysis results showed that the quality of samples from different batches was different. The samples were classified into three clusters by HCA and PCA. The method is simple, sensitive, accurate, and efficient, and can be used for the quality evaluation of C. decapetala.
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Caesalpinia , Chromatography, High Pressure Liquid , Chromatography, Liquid , Principal Component Analysis , Tandem Mass SpectrometryABSTRACT
The fruits of leguminous plants Cercis Chinensis Bunge are still overlooked although they have been reported to be antioxidative because of the limited information on the phytochemicals of C.chinensis fruits.A simple,rapid and sensitive HPLC-MS/MS method was developed for the identification and quantitation of the major bioactive components in C.chinensis fruits.Eighteen polyphenols were iden-tified,which are first reported in C.chinensis fruits.Moreover,ten components were simultaneously quantified.The validated quantitative method was proved to be sensitive,reproducible and accurate.Then,it was applied to analyze batches of C.chinensis fruits from different phytomorph and areas.The principal components analysis (PCA) realized visualization and reduction of data set dimension while the hierarchical cluster analysis (HCA) indicated that the content of phenolic acids or all ten components might be used to differentiate C.chinensis fruits of different phytomorph.
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Identification of the chemical compositionof essential oils is very important for ensuring the quality of finished herbal products. The objective of the study was to analyze the chemical components present in the essential oils of five Beilschmiediaspecies (i.e. B. kunstleri, B. maingayi, B. penangiana, B. madang, and B. glabra) by multivariate data analysis using principal component analysis (PCA) and hierarchical clustering analysis (HCA) methods. The essential oils were obtained by hydrodistillation and fully characterized by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). A total of 108 chemical components were successfully identified from the essential oils of five Beilschmiediaspecies. The essential oils were characterized by high proportions of ß-caryophyllene (B.kunstleri), δ-cadinene (B. penangianaand B. madang), and ß-eudesmol (B. maingayiand B. glabra). Principal component analysis (PCA) and hierarchical cluster analysis (HCA) revealed that chemical similarity was highest for all samples, except for B. madang. The multivariate data analysis may be used for the identification and characterization of essential oils from different Beilschmiediaspecies that are to be used as raw materials of traditional herbal products.
La identificación de la composición química de los aceites esenciales es muy importante para garantizar la calidad de los productos herbales terminados. El objetivo del estudio fue analizar los componentes químicos presentes en los aceites esenciales de cinco especies de Beilschmiedia (B. kunstleri, B. maingayi, B. penangiana, B. madangy B. glabra) mediante análisis de datos multivariados utilizando los métodos de análisis de componente principal (PCA) y análisis de agrupamiento jerárquico (HCA). Los aceites esenciales se obtuvieron por hidrodestilación y se caracterizaron completamente por cromatografía de gases (GC) y cromatografía de gases-espectrometría de masas (GC-MS). Se identificaron con éxito un total de 108 componentes químicos a partir de los aceites esenciales de las cinco especies de Beilschmiedia. Los aceites esenciales se caracterizaron por altas proporciones de ß-cariofileno (B. kunstleri), δ-cadineno (B. penangianay B. madang) y ß-eudesmol (B. maingayiy B. glabra). El análisis de componentes principales (PCA) y el análisis de conglomerados jerárquicos (HCA) revelaron que la similitud química fue más alta para todas las muestras, excepto para B. madang. El análisis de datos multivariados puede usarse para la identificación y caracterización de aceites esenciales de diferentes especies de Beilschmiedia que se utilizan como materias primas de productos herbales tradicionales.
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Oils, Volatile/chemistry , Lauraceae/chemistry , Sesquiterpenes/analysis , Cluster Analysis , Distillation , Multivariate Analysis , Chromatography, Gas/methods , Principal Component Analysis , Monoterpenes/analysisABSTRACT
Objective: Poria cocos and Polyporus umbellatus are similar medicinal fungi in traditional Chinese medicines. A method for fingerprint analysis of monosaccharide composition of polysaccharides by HPLC combined with chemometrics methods has been developed for characterization and discrimination of them in this research. Methods: The polysaccharides were extracted by decocting in water, and then completely hydrolyzed with hydrochloride. Monosaccharides in the hydrolyzates were derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP) for HPLC analysis. More than 20 batches of P. cocos and P. umbellatus from different regions were analyzed. Results: The fingerprints of P. cocos showed five common characteristic peaks, which were identified by comparing with the reference substances. The five peaks corresponded to the derivatives of mannose, ribose, glucose, galactose, and fucose. At the same time, the fingerprints of P. umbellatus showed eight common characteristic peaks, of which seven were identified as the derivatives of mannose, ribose, rhamnose, glucose, galactose, xylose, and fucose. Moreover, the similarity of their fingerprints was respectively calculated by the Similarity Evaluation System for Chromatographic Fingerprint of TCM published by China Pharmacopoeia Committee (Version 2004A). And the data were further processed by hierarchical cluster analysis (HCA) and principal component analysis (PCA). The similarity evaluation and HCA indicated that there were no significant difference in P. cocos or P. umbellatus samples from different geographical regions, but PCA was performed to characterize the difference in monosaccharide constituents between P. cocos and P. umbellatus, and linear discriminant analysis (LDA) showed the overall correct classification rate was 100%. Conclusion: The fingerprint analysis method of monosaccharide composition of water-soluble polysaccharides can distinguish P. cocos and P. umbellatus, and can be applied for the authentication or quality control for P. cocos and P. umbellatus.
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The traditional Chinese medicine of Radix Hedysari plays an important role in invigorating gas for as-cending, benefiting blood for promoting production of fluid, and promoting circulation for removing obstruction in collaterals, which is consistent with the principle of treatment for osteoporosis. This study is designed to investigate the bioactive components on increasing peak bone mass (PBM) by exploring the spectrum-effect relationship between chromatography fingerprints and effect. Multiple indicators are selected to evaluate the pharmacological activity. In fingerprints, 21 common peaks are obtained, five of which are identified. Furthermore, gray relational analysis (GRA) is a quantitative method of gray system theory and is used to describe the correlation degree of common peaks and pharmacological activities with relational value. 21 components are then divided into three different regions, of which ononin and calycosin play an extremely significant role in increasing PBM. In addition, factor analysis and hierarchical cluster analysis (HCA) are used to screen the optimal producing area for Radix Hedysari. This provides a comprehensive and efficient method to improve the quality evaluation of Radix Hedysari, confirming the bioactive components for PBM-enhancement and further develop its medicinal value.
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Objective: The HPLC fingerprints of water extracts and alcohol extracts of Glycyrrhiza uralensis from different producing areas were established in order to provide reference for the quality control and the extraction and processing methods. Methods: The HPLC fingerprints of water extracts and alcohol extracts of 10 batches of G. uralensis from different producing areas were established by HPLC method, and the similarity evaluation and hierarchical cluster analysis of the obtained fingerprint data were carried out. Results: The fingerprint of water extracts and ethanol extracts of G. uralensis was established. There were 16 common peaks were identified from fingerprints of water extracts, and six components were identified, meanwhile, the fingerprints of water extracts showed that the similarity of 10 batches of G. uralensis was in the range of 0.030 and 0.999. Sixty common peaks were identified from fingerprints of ethanol extracts, and seven components were identified, meanwhile, the fingerprints of ethanol extracts showed that the similarity of 10 batches of G. uralensis were all greater than 0.85. The results of hierarchical cluster analysis showed that the water extracts and alcohol extracts of G. uralensis in different areas could be well classified, suggesting that natural factors such as the climate, soil conditions and other natural factors had a significant impact on the internal quality of licorice. Conclusion: A stable and reliable HPLC fingerprint evaluation method for water extracts and alcohol extracts of G uralensis has been established, which can provide a powerful theoretical basis and a guidance for the study of quality standard of G. uralensis and the optimization of extraction and processing technology, and provide a scientific basis for the choice of clinical medication.
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Objective: Crude Leonuri Fructus (CLF), the fruits of the Leonurus japonicus Houtt, and processed Leonuri Fructus (PLF) by stir-baking as the important Chinese herbal medicines, have been used in China and other Asian countries for thousands of years. The objective of this research is to reveal the difference between CLF and PLF. Methods: The sensory technologies of the colorimetry, sensitive and validated HPLC-ELSD and GC combined with flame ionization detector (GC-FID) were employed to discriminate CLF and its processed product PLF. The color parameters of the samples were determined by colorimetric instrument CR-410. Moreover, the content of stachydrine and six fatty acids were determined by HPLC and GC. Subsequently, analysis of variance (ANOVA), principal components analysis (PCA), hierarchical cluster analysis (HCA), and Kendall's correlation test were performed for data analysis. Results: The CLF and PLF were divided into two categories by PCA and HCA in terms of their component content and color. The results distinctly demonstrated significant changes in color and the content of indicative components between CLF and PLF. Conclusion: The study revealed that HPLC, GC, and colorimetric method in combination with chemometric method could be used as comprehensive quality evaluation for CLF and PLF.
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Objective A method for classification and identification of Rhodiola quadrifida and Rhodiola crenulata was established based on nuclear magnetic resonance 1H-NMR fingerprints-chemical pattern recognition technique. Methods Using high resolution (600 MHz) NMR fingerprints pattern technique, the total component information 1H-NMR fingerprint of R. quadrifida and R. crenulata was determinated, combined with the similarity analysis, hierarchical cluster analysis, principal component analysis (PCA), and partial least squares discriminant analysis (PLS-DA) methods for chemical pattern recognition analysis. Results 1H-NMR fingerprints techniques combined with chemical pattern recognition analysis was an effective method to discriminate and identify R. quadrifida and R. crenulata. The difference of the 1H-NMR fingerprint of R. quadrifida and R. crenulata was obvious, which truly and comprehensively reflected the characteristic components and internal qualities of Rhodiola. The main different components of R. quadrifida and R. crenulata were terpenoids and flavonoids, in particular, crenulatin of the terpenoid was a characteristic ingredient in the identification of R. quadrifida and R. crenulata, which can be used as the identification and classification index of R. quadrifida and R. crenulata. Conclusion 1H-NMR fingerprints techniques combined with chemical pattern recognition analysis method is an effective method for classification and identification of Rhodiola, which lays the foundation for variety identification and quality evaluation of medicinal plants of Rhodiola.
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@#Introduction: This study determined nitrate concentration and identify the classifying sources of nitrate pollution in the alluvial deposit aquifer system in Bachok, Kelantan. Materials and Methods: A total of 300 groundwater samples were collected in two different areas; agricultural area (150 samples) and non-agricultural area (150 samples). The samples were analyzed for nitrate and other parameters such as pH, EC, NH4+, TDS, turbidity and salinity. The multivariate analyses were used to identify factors that govern the groundwater quality and potential nitrate sources in the study area. Results: Samples in the agricultural area were slightly acidic (5.89 ± 0.67), contained high nitrate (15.10 ± 15.90 mg/L NO3-N), NH4+ (0.82 ± 1.24 mg/L) and turbidity (3.25 ± 2.78 NTU). The principal component analysis (PCA) have identified the groundwater quality in the study area was influenced by the natural processes and anthropogenic activities. Based on the hierarchal cluster analysis (HCA), Cluster II in the agricultural area was identified to be most heavily nitrate contamination, while Cluster III in the non-agricultural area was identified to be strongly affected by seawater intrusion. Conclusion: The findings of this study are useful for developing protection alternatives of private well waters to prevent further deterioration of groundwater quality by nitrate such as control of nitrogen fertilizer use, manure applications and other agricultural practices in the agricultural area. In order to reduce the health risk of nitrate, private well water users in this area should be advised to treat their water or find alternative sources for drink
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Objective: To compare the contents of phenolic acids and flavonoids in wild grown and cultivated Prunella vulgaris from various habitats, an HPLC method was established for simultaneous determination of five bioactive compounds (rosmarinic acid, salviaflaside, caffeic acid, rutin, and luteolin). Methods: The Agilent 5HC-C18 (250 mm × 4.6 mm, 5 μm) was adopted with a gradient eluent system composed of acetonitrile and 0.1% phosphoric acid aqueous solution at the temperature of 30℃. The flow rate was 1.0 mL/min. The detection wavelength was 280 nm. Independent t-test (t-test), hierarchical clustering analysis (HCA), and correlation analysis were applied to analyzing and evaluating wild grown and cultivated P. vulgaris. Results: The t-test results showed that the contents difference of rosmarinic acid, rutin, caffeic acid, and luteolin had statistical significance (P 0.05). According to the result of HCA, most of the cultivated and wild materials could be differentiated. The result of correlation analysis showed that the ear length has no significant influence on the contents of main compounds in P. vulgaris. Conclusion: The determination method is simple and feasible, and it can be used as one of the quality evaluation method of P. vulgaris.
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A new method was developed for the chromatographic fingerprint analysis of Toosendan Fructus by HPLC coupled with the charged aerosol detector (CAD) in the present study. Samples were well separated on an Agilent ZOBAX SB C18 column (4.6 mm×250 mm, 5 μm) by gradient elution using acetonitrile and water containing 0.1% formic acid (v/v) at the flow rate of 1.0 mL·min-1. The column temperature was 30℃ and the injection volume was 5 μL. The nitrogen inlet pressure of the charged aerosol detector (CAD) was 35 psi, and the nebulizer chamber temperature was 35℃. In addition, the method of the chromatographic fingerprints combined with multivariate statistical analysis was effective and reasonable lead to an accurate classification of 20 batches of samples from different locations. The results showed that 28 common peaks were observed in the fingerprint and the samples were classified into three clusters. The established method was well validated, and showed high precision, good repeatability, and satisfactory stability. It may serve in the quality control and evaluation of Toosendan Fructus.
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OBJECTIVE: To establish a rapid approach for quality evaluation of Huoxiang Zhengqi Tincture. METHODS: Fingerprint was established by ultra performance liquid chromatography (UPLC) method and method evaluation was performed. By comparing with reference substances, reference drugs and reference extractives, the characteristic peaks and their ascriptions were investigated. Twenty-eight batches of samples from 13 manufactures were determined and their fingerprints were compared. Then two dimensional hierarchical cluster analysis (2D-HCA) and principal analysis (PCA)were applied in pattern recognition. RESULTS: Eighteen characteristic peaks were mainly related to Magnoliae Officinalis Cortex, Citri Pericarpium Reticulatae, Angelicae Dahuricae Radix, Extractum Glycyrrhizae, Atractylodis Rhizoma and patchouli oil. The UPLC fingerprints of samples from different manufacturers differed a lot. The results of 2D-HCA and PCA were consistent, which indicated that the fingerprints of the samples from the same manufacturer could cluster to their own group and the characteristic peaks of honokiol, magnolol and hesperidin showed the greatest impact on the fingerprints. CONCLUSION: UPLC fingerprinting with aid of chemometric analysis is rapid, simple, objective and easy to realize digitalization, thus providing a novel way for quality evaluation of Huoxiang Zhengqi Tincture.
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Abstract Ipanema National Forest, southeastern Brazil, once contained 340 bird species. Forest cover suffered for centuries from log exploitation and, as a result, most of the remaining forests are now an impoverished subset of the original vegetation. We show how the bird community changed over time by comparing historical and recent records. Currently, 228 species can be recorded, for a compilation of 410 species, of which 359 are documented. Some 89 forest species with historical records failed to be detected in recent surveys. Of the 72 Atlantic Forest or Cerrado endemic species, no more than 29 (40%) are still found. The bird community changed from one which used to be related to coastline rain forests to another, which relates more to drier semideciduous forests of the interior.
Resumo A Floresta Nacional de Ipanema, sudeste do Brasil, já abrigou 340 espécies de aves. Sua cobertura florestal sofreu por séculos com a exploração de madeira e, desse modo, a maior parte da vegetação remanescente é uma sub-representação daquela original. Neste artigo é demonstrado como a comunidade de aves foi modificada com o passar do tempo por meio da comparação entre registros históricos e recentes. Atualmente, 228 espécies podem ser registradas, para um total de 410 espécies, das quais 359 possuem documentação. Das espécies registradas historicamente, 89 não foram mais detectadas. Das 72 espécies endêmicas da Mata Atlântica ou do Cerrado, apenas 29 (40%) ainda podem ser encontradas. A comunidade de aves, outrora similar à de florestas ombrófilas costeiras, atualmente é mais relaciona à comunidade de matas semideciduais mais secas do interior.
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ABSTRACT Plants from the Momordica genus, Curcubitaceae, are used for several purposes, especially for their nutritional and medicinal properties. Commonly known as bitter gourds, melon and cucumber, these plants are characterized by a bitter taste owing to the large content of cucurbitacin compounds. However, several reports have shown an undisputed correlation between the therapeutic activities and polyphenolic flavonoid content. Using ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry in combination with multivariate data models such as principal component analysis and hierarchical cluster analysis, three Momordica species (M. foetida Schumach., M. charantia L. and M. balsamina L.) were chemo-taxonomically grouped based on their flavonoid content. Using a conventional mass spectrometric-based approach, thirteen flavonoids were tentatively identified and the three species were found to contain different isomers of the quercetin-, kaempferol- and isorhamnetin-O-glycosides. Our results indicate that Momordica species are overall very rich sources of flavonoids but do contain different forms thereof. Furthermore, to the best of our knowledge, this is a first report on the flavonoid content of M. balsamina L.
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To compare the main nucleosides in Cordyceps genus herbs (C. sinensis, C. millitaris, Hirsutella sinensis and C. sobolifera), an HPLC method for simultaneous determination of uridine, inosine, guanosine, adenosine and cordycepine in Cordyceps genus herbs was developed. The sample was extracted with 0.5% phosphoric acid solution to prepare test solution. The separation was performed on a Zorbax SB-Aq (4.6 mm×150 mm, 5 μm) column with gradient elution by 0.04 mol•L⁻¹ potassium dihydrogen phosphate solution and acetonitrile, column temperature 30 ℃,flow rate 0.8 mL•min⁻¹,and detection wavelength 260 nm. The content of nucleosides in four Cordyceps genus herbs was evaluated by fingerprint analysis and hierarchical cluster analysis (HCA). The calibration curves of five nucleosides showed good linear regression (r>0.99) and the average recoveries were between 95.0% and 105.0%. The contents of the five nucleosides in the four Cordyceps genus herbs were different and could be obviously distinguished by HCA. The fingerprint analysis result showed that the similarity between C. sinensis and the others was less than 0.9. The method was accurate and reliable, which can be used for quality control of Cordyceps genus herbs.
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Objective To study the distinct clinical phenotype of chronic airway diseases by hierarchical cluster analysis and two-step cluster analysis.Methods A population sample of adult patients in Donghuamen community,Dongcheng district and Qinghe community,Haidian district,Beijing from April 2012 to January 2015,who had wheeze within the last 12 months,underwent detailed investigation,including a clinical questionnaire,pulmonary function tests,total serum IgE levels,blood eosinophil level and a peak flow diary.Nine variables were chosen as evaluating parameters,including pre-salbutamol forced expired volume in one second(FEV1)/forced vital capacity (FVC) ratio,pre-salbutamol FEV1,percentage of post-salbutamol change in FEV1,residual capacity,diffusing capacity of the lung for carbon monoxide/alveolar volume adjusted for haemoglobin level,peak expiratory flow (PEF) variability,serum IgE level,cumulative tobacco cigarette consumption (pack-years) and respiratory symptoms (cough and expectoration).Subjects' different clinical phenotype by hierarchical cluster analysis and two-step cluster analysis was identified.Results (1) Four clusters were identified by hierarchical cluster analysis.Cluster 1 was chronic bronchitis in smokers with normal pulmonary function.Cluster 2 was chronic bronchitis or mild chronic obstructive pulmonary disease (COPD) patients with mild airflow limitation.Cluster 3 included COPD patients with heavy smoking,poor quality of life and severe airflow limitation.Cluster 4 recognized atopic patients with mild airflow limitation,elevated serum IgE and clinical features of asthma.Significant differences were revealed regarding pre-salbutamol FEV1/FVC%,pre-salbutamol FEV1% pred,postsalbutamol change in FEV1 %,maximal mid-expiratory flow curve (MMEF)% pred,carbon monoxide diffusing capacity per liter of alveolar(DLCO)/(VA)% pred,residual volume(RV)% pred,total serum IgE level,smoking history (pack-years),St.George' s respiratory questionnaire (SGRQ) score,acute exacerbation in the past one year,PEF variability and allergic dermatitis (P < 0.05).(2) Four clusters were also identified by two-step cluster analysis as followings,cluster 1,COPD patients with moderate to severe airflow limitation;cluster 2,asthma and COPD patients with heavy smoking,airflow limitation and increased airways reversibility;cluster 3,patients having less smoking and normal pulmonary function with wheezing but no chronic cough;cluster 4,chronic bronchitis patients with normal pulmonary function and chronic cough.Significant differences were revealed regarding gender distribution,respiratory symptoms,pre-salbutamol FEV1/FVC%,pre-salbutamol FEV1 % pred,post-salbutamol change in FEV1 %,MMEF% pred,DLCO/VA% pred,RV% pred,PEF variability,total serum IgE level,cumulative tobacco cigarette consumption (pack-years),and SGRQ score (P < 0.05).Conclusion By different cluster analyses,distinct clinical phenotypes of chronic airway diseases are identified.Thus,individualized treatments may guide doctors to provide based on different phenotypes.