Chemical constituents from Callicarpa kwangtungensis and their hemostatic activities / 中国中药杂志

This study aims to identify the chemical constituents from Callicarpa kwangtungensis and determine their activities. MCI, ODS, and Sephadex LH-20 chromatography and semi-preparative HPLC were employed to separate the chemical constituents. A total of 15 compounds were separated, and their structures were identified on the basis of spectroscopic analysis and comparison with the data in relevant literature. Specifically, the 15 compounds were 3-O-α-L-rhamnopyranosyl-6-O-β-D-apiofuranosyl-4-O-E-caffeoyl-D-glucopyranoside(1), 3,6-O-α-L-dirhamnopyranosyl-4-O-E-caffeoyl-D-glucopyranoside(2), β-OH-forsythoside B(3), β-OH-poliumoside(4),(+)-lyoniresinol-3α-O-β-D-apiofuranosyl-(1→2)-β-D-glucopyranoside(5),(+)-lyoniresinol-3α-O-β-D-glucopyranoside(6),(-)-lyoniresinol-3α-O-β-D-glucopyranoside(7), kelampayoside A(8), descaffeoylpoliumoside(9), acteoside(10), alyssonoside(11), poliumoside(12), isacteoside(13), acetyl forsythoside B(14), and forsythoside B(15). Compounds 1 and 2 were novel, and the NMR data of compounds 3 and 4 were reported here for the first time. Furthermore, the hemostatic activities of the extract and abundant ingredients(compounds 12 and 15) of C. kwangtungensis were determined with Yunnan Baiyao as the positive control and normal saline as the negative control. The extract and compounds 12 and 15 significantly shortened the tail tip bleeding time in mice.

A new labdane-type diterpenoids from Callicarpa nudiflora / 中国中药杂志

The purpose of the research is to study the bioactive constituents of Callicarpa nudiflora. From the 65% ethanol extract of C. nudiflora leaves, ten compounds were isolated by macroporous adsorption resin, Sephadex LH-20, ODS, silica gel, and preparative HPLC. These compounds were identified as callicapene M6(1), sterebin A(2), isomartynoside(3), crenatoside(4), luteolin-7-O-neohesperidoside(5), apigenin-7-O-β-D-neohesperidoside(6), isoacteoside(7), acteoside(8),(7R)-campneoside I(9), and(7S)-campneoside I(10) on the basis of NMR, HR-ESI-MS, and optical rotation data. Compound 1 was obtained as a new compound. Compounds 2 and 4 were isolated from the genus Callicarpa for the first time. Compounds 9 and 10 were isolated from C. nudiflora for the first time.

3, 4-seco-Isopimarane and 3, 4-seco-pimarane diterpenoids from Callicarpa nudiflora / 中国天然药物

A phytochemical investigation was carried out on the extract of a medicinal plant Callicarpa nudiflora, resulting in the characterization of five new 3, 4-seco-isopimarane (1-5) and one new 3, 4-seco-pimarane diterpenoid (6), together with four known compounds. The structures of the new compounds were fully elucidated by extensive analysis of MS, 1D and 2D NMR spectroscopic data, and time-dependent density functional theory (TDDFT) calculation of electronic circular dichroism (ECD) spectra, and DFT calculations for NMR chemical shifts and optical rotations.

Comparative study on distribution of Callicarpa nudiflora by different regionalization methods / 中国中药杂志

This article is based on basic data such as field surveys and literature surveys, contrasting and analyzing the distribution of Callicarpa nudiflora by different zoning methods, different data sources, and different spatial scales. The results showed that there were certain differences in the distribution results obtained by using different methods, such as qualitative description, similar ecological environment, and niche model, to divide the distribution of the C. nudiflora, but all of them could reflect the distribution of C. nudiflora to different degrees. Among them, the qualitative description division method has certain advantages in macro guidance in a large scale. The distribution range obtained by the ecological environment similar division method is wider than that obtained by applying the qualitative description method and the niche model method. The results of the zoning of the distribution of the C. nudiflora obtained from different data sources were different. The number and representativeness of the survey data have an impact on the zoning results. Through the analysis of the distribution of different spatial scales, the ecological factors and contribution rates that affect the distribution of C. nudiflora are different in China and in the world. The comprehensive multi-source data analysis showed that C. nudiflora mainly distributed in southern coastal provinces such as Hainan, Guangdong, Guangxi and Fujian in China, and also in Jiangxi, Guizhou, Yunnan, Sichuan, Chongqing, Hunan, Gansu, Taiwan and other provinces. Globally, C. nudiflora are suitable for distribution in Southeast Asia, such as China, Vietnam, Laos, Myanmar, India, etc. There are also potential distribution areas in the southern United States and Mexico.

Genome survey and characteristic analysis of SSR in Callicarpa nudiflora / 中国中药杂志

Callicarpa nudiflora,which is a big brand of Li nationality medicine with Hainan characteristics,has the effects of dissolving stasis,hemostasis,anti-inflammatory and antibacterial. At present,there is a lack of information about the reference genome of C. nudiflora. The study of the genome size,heterozygosity rate and characteristics of SSR of C. nudiflora,can provide an effective basis for the formulation of the whole genome de novo sequencing strategy and development of SSR molecular markers of C. nudiflora. To realize this purpose,high throughput sequencing platform Illumina Hiseq was used to sequence the genome structure of C. nudiflora and K-mer analysis was applied to estimate genome size,repeat sequences and heterozygosity rate. Simple-sequence repeat( SSR) loci that are suitable as markers were identified by MISA software. The results showed the estimated genome size of C. nudiflora was 822. 43 Mb,with a 0. 85% heterozygosity rate and 71. 67% repeats,and the GC content of genome was about 49. 20%. Therefore,C. nudiflora belongs to a complex genome with high heterozygosity and repetition. SSR molecular genetic markers were analyzed in the genome sequence,and a total of 206 049 SSRs were identified,among which mono-nucleotide,di-nucleotide and tri-nucleotide repetitive motifs summed up to 198 993,accounting for 96. 57% of the total SSRs. Among the 2-6 nucleotide repeats,AT/AT,AAT/ATT,AGCC/CTGG,AAAAT/ATTTT and AGATAT/ATATCT have the largest number,respectively. This report represents the first genome-wide characterization of C. nudiflora,and provides a reference for the construction of the library for the fine sequencing of the genome,and a molecular basis for the development of SSR molecular markers as well as for the protection and utilization of gene resources.

Application of UAV remote sensing in monitoring of Callicarpa nudiflora / 中国中药杂志

In order to solve the problem of manual area measurement,the traditional methods of medicinal planting area statistics are difficult to meet the needs of rapid area survey application. This paper uses the UAV remote sensing method with the advantages of unmanned,automatic,high efficiency,high score and short production cycle to monitor the shape of Callicarpa nudiflora. A solution for aerial photography,image data acquisition and data processing of drones were designed for characteristics and planting conditions. After data processing and statistical analysis,detailed information on the location and area of the C. nudiflora in the target area was obtained. Then the accuracy comparison analysis was carried out with the measured results of the C. nudiflora. The results show that the UAV is feasible for the monitoring of C. nudiflora,and has a good application prospect in the monitoring of Chinese herbal medicine planting.

Random forest classification of Callicarpa nudiflora from WorldView-3 imagery based on optimized feature space / 中国中药杂志

Taking the Xiushui township of Baisha county in Hainan province as the research area,the random forest algorithm with obvious advantages in feature selection and classification extraction was used to extract the information of the Callicarpa nudiflora planting in the study area. Firstly,four kinds of different characteristic variables were generated based on World View-3 data,including spectral features,principal component features,vegetation index and texture features. Secondly,the spatial distribution of the C. nudiflora in the study area was extracted by remote sensing by random forest classification algorithm. Finally,the feature space of the random forest classification algorithm was optimized based on the feature importance to obtain the best random forest classification results,and this result is compared with the classification result of the random forest algorithm of the unoptimized feature space. The results showed that:①The overall accuracy of the C. nudiflora extracted by World View-3 image was 89. 97%,and the Kappa coefficient was 0. 84,which indicates that the random forest algorithm had higher classification accuracy and better applicability in Hainan C. nudiflora recognition.② The overall accuracy of extracting C. nudiflora with the dimension reduction feature was 90. 4,and the Kappa coefficient was 0. 85,which indicates that the random forest algorithm can effectively select features. At the same time as the feature variable data mining,the precision of the information extraction of the C. nudiflora was still guaranteed,and the operation efficiency was improved. This study provides a new idea,method and technical means for information extraction of cultivated medicinal plant resources in terms of feature selection and method selection.

Analysis of constituents from different parts of Callicarpa nudiflora by UPLC-Q-TOF-MS/MS / 中国中药杂志

The aim of this study was to clarify the main components of the green leaves of Callicarpa nudiflora,and to compare the difference of main components between the green leaves,yellow leaves,branches and seeds. In this study,ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) coupled with the UNIFI scientific information system was adopted. And the identification of the main chemical components of C. nudiflora was combined with reference materials,literatures and online database. In addition,the difference of main components was analyzed by Progenesis QI,principal component analysis(PCA) and orthogonal partial least squares discriminant(OPLS-DA). A total of 57 compounds were identified in green leaves,including phenylpropanoids,flavonoids and iridoids. Among them,the relative content of phenylethanoid glycosides was highest. Furthermore,the PCA analysis showed that there are significant differences in main components of the branches and other parts of C. nudiflora. Combined with OPLS-DA analysis,nudifloside,parvifloroside B and β-hydroxysamioside were selected as the characteristic components for distinguish the leaves and branches of C. nudiflora. Our study provided a scientific basis for the collection and identification of C. nudiflora.

Identification of poliumoside metabolites in rat plasma, urine, bile, and intestinal bacteria with UPLC/Q-TOF-MS / 中国天然药物

Poliumoside is representative of phenylethanoid glycosides, which are widely found in many plants. Poliumoside is also regarded as the main active component of Callicarpa kwangtungensis Chun (CK), though its oral bioavailability in rat is extremely low (0.69%) and its in vivo and in vitro metabolism has not yet been systematically investigated. In the present study, an ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS) method was employed to identify the metabolites and investigate the metabolic pathways of poliumoside in rat after oral administration 1.5 g·kg of poliumoside. As a result, a total of 34 metabolites (30 from urine, 17 from plasma, and 4 from bile) and 9 possible metabolic pathways (rearrangment, reduction, hydration, hydrolyzation, dehydration, methylation, hydroxylation, acetylation, and sulfation) were proposed in vivo. The main metabolite, acteoside, was quantified after incubated with rat intestinal bacteria in vitro. In conclusion, the present study systematically explored the metabolites of poliumoside in vivo and in vitro, proposing metabolic pathways that may be significant for further metabolic studies of poliumoside.

Identification of poliumoside metabolites in rat plasma, urine, bile, and intestinal bacteria with UPLC/Q-TOF-MS / 中国天然药物

Poliumoside is representative of phenylethanoid glycosides, which are widely found in many plants. Poliumoside is also regarded as the main active component of Callicarpa kwangtungensis Chun (CK), though its oral bioavailability in rat is extremely low (0.69%) and its in vivo and in vitro metabolism has not yet been systematically investigated. In the present study, an ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS) method was employed to identify the metabolites and investigate the metabolic pathways of poliumoside in rat after oral administration 1.5 g·kg of poliumoside. As a result, a total of 34 metabolites (30 from urine, 17 from plasma, and 4 from bile) and 9 possible metabolic pathways (rearrangment, reduction, hydration, hydrolyzation, dehydration, methylation, hydroxylation, acetylation, and sulfation) were proposed in vivo. The main metabolite, acteoside, was quantified after incubated with rat intestinal bacteria in vitro. In conclusion, the present study systematically explored the metabolites of poliumoside in vivo and in vitro, proposing metabolic pathways that may be significant for further metabolic studies of poliumoside.

Study on HPLC fingerprint of Callicarpa nudiflora and determination of ten components / 中国中药杂志

This study is to establish the HPLC fingerprint and determine eight components of Callicarpa nudiflora, and provide a scientific basis for the identification and quality control. The Waters sunfire C18 column (4.6 mm x 250 mm, 5 µm) was used and the detection wavelength was 330 nm . The column temperature was 30 °C. The mobile phases were acetonitrile (A) and 0.1% formic acid (B) eluting in a gradient program at a flow rate of 1.0 mL · min(-1). The chromatographic fingerprint similarity evaluation system for tradition Chinese medicine(2012) was used for analysis. C. nudiflora from different samples were of high similarity in fingerprint and the separation of ten components was good. There was an obvious difference between other samples and C. nudiflora leaves. In quantitative analysis, the ten components showed good regression(R2 > 0 999 0) with linear ranges, and their recoveries were in the range of 96.0%-105.0%. The established qualitative and quantitative methods are highly specific, simple and accurate, which can be used for the identification and quality control of C. nudiflora.

Chemical constituents from Callicarpa nudiflora and their cytotoxic activities / 中国中药杂志

The chemical consitituents from cytotoxic fraction of the Callicarpa nudiflora extract were isolated and purified by a combination of HP-20 macroporous resin, silica gel and Sephadex LH-20 column chromatographies. The structures were elucidated on the basis of the spectroscopic data and comparison of their spectroscopic data with reported data. The cytotoxicity was evaluated by the MTT assay. The 50% and 70% EtOH elutions of EtOH-extract showed significant cytotoxic activities, leading to the isolation of twelve compounds, which were identified as luteoloside(1), lutedin-4'-O-β-D-glucoside(2), 6-hydroxyluteolin-7-O-β-glucoside(3), lutedin-7-O-neohesperidoside(4), rhoifolin (5), luteolin-7, 4'-di-O-glucoside (6), forsythoside B (7), acteoside (8), alyssonoside (9), catalpol(10), nudifloside(11), and leonuride(12). Compounds 3-6, 10 and 12 were isolated from this genus for the first time, and compound 9 was isolated from this plant for the first time. The cytotoxicity assay demonstrated that flavonoids 1-6, in various concentrations, showed monolithic proliferation inhibitory activities against Hela, A549 and MCF-7 cell lines. Compounds 3, 5 and iridoid glycoside 11 possessed higher cytotoxicacivities. In short, flavonoids are the main components of cytotoxic extract from C. nudiflora, while phenylethanoid glycosides are the predominant ingredient but inactive to cancer cell lines. In addition, the minor iridoid glycoside expressed weak cytotoxic activity.

Determination of forsythiaside B and poliumoside in different origin and parts from Callicarpa kwangtungensis / 中国中药杂志

The RP-HPLC method was used to determinate the contents of forsythiaside B and poliumoside in different origin and parts from Callicarpa kwangtungensis. The linear ranges of forsythiaside B and poliumoside were 0. 106-3. 18 and 0. 105 2-3. 156 microg, respectively. The average recoveries of forythiaside B and poliumoside were 99. 01% ( RSD 1. 2%) and 100. 13% (RSD 0. 90% ), respectively. The contents of forsythiaside B and poliumoside were changed in different origin and parts from C. kwangtungensis. The sample from the area of Luxi, Pingxiang City, Jiangxi Province has the highest contents of forsythiaside B and poliumoside. The contents of forsythiaside B and poliumoside in different parts from C. kwangtungensis in Luxi are: leaf > stem > fruit. This result will provide a scientific basis for quality control and reasonable utilzation of C. kwangtungensis.

Triterpenes from Callicarpa integerrima Champ / 药学学报

A new triterpenoid saponin and fourteen known triterpenoids were isolated from the methanol extract of the stems and leaves of Callicarpa integerrima Champ, which is used in Chinese folk medicine for stopping bleeding, expelling the wind, dissipating stagnation, and treating scrofula, by using various chromatographies, such as silica gel, Sephadex LH-20 and RP-C18 column chromatography. Their structures were identified as a new compound 2alpha, 3beta, 19alpha, 23-tetrahydroxy-olean-12-en-28-oic acid-28-O-beta-D-glucopyranosyl-(1 --> 4)-beta-D-glucopyranoside (1), together with fourteen known compounds: oleanolic acid (2), 3-acetyl oleanolic acid (3), 3beta-O-acetyl ursolic acid (4), 2alpha-hydroxy-ursolic acid (5), 2alpha, 3beta, 19alpha, 23-tetrahydroxy-urs-12-en-28-oic acid (6), alpha-amyrin-3-O-beta-D-glucopyranoside (7), pomolic acid (8), betulinic acid (9), ursolic acid (10), 2alpha, 3beta, 19alpha, 23-tetrahydroxy-olean-12-en-28-oic acid (arjungenin) (11), 2alpha-hydroxy-oleanolic acid (12), hederagenin (13), 2alpha, 19alpha-dihydroxy-ursolic acid (14) and pruvuloside A (15), by the spectroscopic techniques of NMR, HMBC, IR and MS, separately. All these compounds were obtained from this plant for the first time, and compounds 3, 4 and 15 were isolated from genus Callicarpa L. for the first time.

Chemical constituents from Callicarpa nudiflora and their hemostatic activity / 中国中药杂志

<p><b>OBJECTIVE</b>To study the hemostatic effect of chemical constituents from Callicarpa nudiflora.</p><p><b>METHOD</b>The chemical constituents were isolated and purified via silica gel and Sephadex LH-20 column chromatography. Their structures were determined on the basis of spectral analysis. prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen (FIB) of the constituents rabbit blood samples were tested with rabbit blood in vitro.</p><p><b>RESULT</b>Eleven compounds were isolated and identified as two diterpenens: 7alpha-hydroxy sandaracopimaric acid (1), 16, 17-dihydroxy-3-oxophyllocladane (2). Two phenoic glycosides: acteoside (3), samioside(4). Three triterpenes: 2alpha, 3alpha, 24-trihydroxy-ursa-12-en-28-oic acid (5), 2alpha, 3alpha, 19alpha-trihydroxyursa-12-en-28-oic acid-28-0-beta-D-glucopyranosyl ester (6), and 2alpha, 3alpha, 19alpha, 23-tetrahydroxy-ursa-12-en-28-oic acid-28-0-beta-D-glucopyranosyl ester (7). Four flavones: rhamnazin (8), 5-Hydroxy-3, 7, 4'-trimethoxy-flavone (9) , 5-Hydroxy-3, 7, 3', 4'-tetramethoxyflavone (10), and luteoloside (11). All Compounds cannot significantly shorten the PT (P < 0.01), compounds 3, 4, 7, 10 can remarkedly increase APTT (P < 0.01), compound 5 can prolong the T( P < 0.01) obviously, and compound 8 can significantly increase the contents of FIB (P < 0.01).</p><p><b>CONCLUSION</b>Compounds 2, 4 and 10 were isolated from this genus for the first time, and compounds 1, 3, 5, 6, 7 and 9 had been isolated from this plant for the first time. The hemostatic effect of C. nudiflora may be related to the activation of the intrinsic blood coagulation system.</p>

RP-HPLC determination of betulinic acid in Callicarpa macrophylla / 中国中药杂志

<p><b>OBJECTIVE</b>To establish an RP-HPLC method for determination of betulinic acid in Callicarpa macrophylla, a commonly used herbal in Yunnan.</p><p><b>METHOD</b>A Kromasil-C18 column (4.6 mm x 200 mm, 5 microm) and a mobile phase consisted of acetonitrile-0.1% phosphoric acid (63: 37) were used. The flow rate was 1.0 mL x min(-1) and the UV detector wavelength was 205 nm.</p><p><b>RESULT</b>Betulinic acid was well separated from other compounds in C. macrophylla. The content of betulinic acid in C. macrophylla from different origins showed apparent differences, the content of betulinic acid in C. macrophylla from Yunnan was the highest. The calibration curve was linear in the range of 0.016 6-0.332 mg x mL(-1) of betulinic acid with correlation coefficient 0.999 8. The average recovery of betulinic acid was 98.5%.</p><p><b>CONCLUSION</b>The method is simple, accurate and reproducible, and is suitable for the quality control of C. macrophylla.</p>