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In this study, we evaluated several agronomic characteristics of S. divaricata when grown in various regions in Vietnam, including Northeast, Northwest, and Central Highlands. Among the three regions, S. divaricata grown in Northwestern Vietnam has the highest yield reaching 6.21 tons/ha, with a total active ingredient content of 0.655% (Prim-O-glucosylcimifugin 0.383% and 5-O-methylvisamminoside 0.272%). This is followed by the Central Highlands of which S. divaricata yielded 4.12 tons/ha, with a total active ingredient content is 0.543% (Prim-O-glucosylcimifugin 0.292% and 5-O-methylvisamminoside 0.251%). The lowest yield of S. divaricata was recorded in Northeast with 3.13 tons/ha, of which a total active ingredient content was 0.394% with 0.253% for Prim-O-glucosylcimifugin and 0.141% for 5-O- methylvisamminoside. With the applied analytical conditions, HPLC - DAD chromatograms are obtained with sharp peaks of prim-O-glucosylcimifugin (tR = 7.51 min) and 5-O-methylvisamminoside (tR = 20.23 min) , balanced, clear on the background of the medicinal plants Saposhnikovia divaricata (Turcz.) Schischk. In particular, the applicable analytical conditions allow for simultaneous qualitative and quantitative analysis of both prim-O-glucosylcimifugin and 5- O-methylvisaminoside. The results of building the standard curve prim-O-glucosylcimifugin Y = (16146)X – 4020.3, R2 = 0.9992, standard curve 5-O- methylvisamminoside Y = (20490)X – 6921.8, R2 = 0.9999. The quantitative results of prim-O-glucosylcimifugin and 5-O- methylvisamminoside in all regions were slightly higher than that of the pharmacopoeias of Vietnam, China and Hong Kong with the total active ingredient content not less than 0.24%. Thus, The study concluded that Vietnam is a country that can develop medicinal plants Saposhnikovia divaricata (Turcz.) Schischk
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Saposhnikovia divaricata (Turcz.) Schischk (S. divaricata, Fangfeng) is a herb in the Apiaceae family, and its root has been used since the Western Han Dynasty (202 B.C.). Chromones and coumarins are the pharmacologically active substances in S. divaricata. Modern phytochemical and pharmacological studies have demonstrated their antipyretic, analgesic, anti-inflammatory, antioxidant, anti-tumor, and anticoagulant activities. Technological and analytical strategy theory advancements have yielded novel results; however, most investigations have been limited to the main active substances-chromones and coumarins. Hence, we reviewed studies related to the chemical composition and pharmacological activity of S. divaricata, analyzed the developing trends and challenges, and proposed that research should focus on components' synergistic effects. We also suggested that, the structure-effect relationship should be prioritized in advanced research.
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Médicaments issus de plantes chinoises/pharmacologie , Coumarines/pharmacologie , Apiaceae/composition chimique , 4H-1-Benzopyran-4-onesRésumé
OBJECTIVE@#Early bolting of Saposhnikovia divaricata has seriously hindered its medicinal value and sustainable development of resources. The molecular mechanism of bolting and flowering of S. divaricata is still unclear and worth of research. In our study, we explored the transcriptome of the genes related to the bolting and flowering of S. divaricata.@*METHODS@#The transcriptome library was constructed, sequenced, assembled and annotated from the bolting and unbolting leaves of S. divaricata by high-throughput sequencing at the bud and flowering stage. Focus on the pathways related to bolting and flowering in plants, and exploring genes. The expression of seven candidate genes was verified by real-time fluorescence quantitative PCR (qRT-PCR).@*RESULTS@#Transcriptome results showed that 249 889 422 high-quality clean reads were obtained. A total of 67 866 unigenes were assembled with an average length of 948.1 bp. Trinity de Novo assembly produced 67 866 unigenes with an average length of 948.1 bp. Among 993 differentially expressed genes, 484 genes were significantly up-regulated and 509 genes were down-regulated in the SdM group. A total of 79 GO terms were significantly enriched for differentially expressed genes. KEGG results showed that 11 154 unigenes were enriched in 89 pathways. And 21 candidate genes related to bolting and flowering of S. divaricata were excavated. The qRT-PCR results showed that expression trends of HDA9, PHYB, AP2, TIR1, Hsp90, CaM, and IAA7 were consistent with transcriptomic sequencing results. In addition, RNA-seq had identified 10 740 transcription factors and classified them into 58 families by their conserved domains. Further studies showed that the transcription factors regulating the flowering of S. divaricata were mainly distributed in the NAC, MYB_related, HB-other, ARF, and AP2 families.@*CONCLUSION@#Based on the results of this study, it was found that the plant hormone signal transduction pathway was one of the decisive factors to control bolting and flowering. Among them, auxin related genes IAA and TIR1 are the key genes in the bolting and flowering process of S. divaricata.
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Objective: Saposhnikoviae Radix (Fangfeng in Chinese), the roots of Saposhnikovia divaricata, lacks commodity specification and grade standardization in the current market. This study investigated the existing specifications and grades of Saposhnikoviae Radix to provide a standardized scientific reference for its market use. Methods: Based on a textual research of Chinese herbal medicine from the Han Dynasty to the present, medicinal materials of different specifications and grades obtained from Saposhnikoviae Radix in the main producing areas of China were collected and the markets for these materials were investigated. Field investigations were performed in the major producing areas such as Northeast China, Hebei Province, and Inner Mongolia. Four major Chinese herbal medicine markets in China were investigated. Sensory indices were used to categorize the two specifications (wild and cultivated) according to the shape, color, texture, and cross-section. High-performance liquid chromatography was performed to determine the active components. Vernier calipers and measuring tape were used to measure the diameter and length, respectively, of 41 samples. Using Excel and the R Language software, cluster analysis and descriptive statistical analysis were performed to assist in the application of new specifications and grades based on physical characteristics, pharmacological activity, and chemical composition. Results: The two specifications (wild and cultivated) of Saposhnikoviae Radix were divided into three grades each based on the length and diameter. Prim-O-glucosylcimifugin, 5-O-methylvisamminoside, and the length of Saposhnikoviae Radix can be used as a basis for classifying the commodity specifications and grades. The specifications and grade standards of Saposhnikoviae Radix were established based on the following eight aspects: shape, surface characteristics, texture, cross section, taste, prim-O-glucosylcimifugin content, 5-O-methylvisamminoside content and length. Conclusion: The formulation of this standard stipulates the commodity specification level of Saposhnikoviae Radix. It is also suitable for the evaluation of commodity specifications in the process of production, circulation and use of Saposhnikoviae Radix.
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Saposhnikovia divaricata (Turcz.) Schischk., a perennial herb belonging to the family Umbelliferae, is widely distributed in Northeast Asia. Its dried root (Radix Saposhnikoviae) is used as a Chinese herbal medicine for the treatment of immune system, nervous system, and respiratory diseases. Phytochemical and pharmacological studies have shown that the main constituents of S. divaricata are chromones, coumarins, acid esters, and polyacetylenes, and these compounds exhibited significant anti-inflammatory, analgesic, antioxidant, antiproliferative, antitumor, and immunoregulatory activities. The purpose of this review is to provide comprehensive information on the botanical characterization and distribution, traditional use and ethnopharmacology, phytochemistry, and pharmacology of S. divaricata for further study concerning its mechanism of action and development of better therapeutic agents and health products from S. divaricata.
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OBJECTIVE： To analyze chemical components as coumarin in Saposhnikovia divaricata， and to provide reference for comprehensive analysis of pharmacodynamic material base in S. divaricata. METHODS： UPLC-Q-TOF-MS method was adopted. Chromatographic condition： the determination was performed on ACQUITY UPLC BEH C18 column with mobile phase consisted of 0.1％ formic acid water-0.1％ formic acid acetonitrile （gradient elution） at the flow rate of 0.3 mL/min. The column temperature was 35 ℃， and sample size was 3 μL. MS condition： ESI， in positive and negative ion mode， ESI+/ESI－ 5 500 V/－4 500 V， declustering potential of 80-－80 V， auxiliary heating gas pressure of 55.00 psi， atomizing gas pressure of －55 psi， curtain gas pressure of －35.00 psi， desolvent temperature of 550 ℃， collision activation scanning energy of 15 eV， collision voltage of 35 psi. Component analysis was performed by comparing with the related literature data and the standard chromatogram control combined with accurate relative molecular mass of compounds provided by UPLC-Q-TOF-MS. RESULTS： Totally 135 chemical components were analyzed in ESI+ mode， and the 105 chemical components were analyzed in ESI－ mode. 11 chemical components as coumarin were identified in ESI+ mode， such as isoimperatorin， umbelliferone， scopolamine， xanthotoxin， psoralen， Ostenol， fraxidin， isoimperatorin， 5-hydroxyl-8-methoxypsoralen， phellopterin， decursin. CONCLUSIONS： The method is accurate and rapid， and can be used for the analysis of chemical components as coumarin in S. divaricata.
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This study aims to compare the internal chemical composition and appearance indifferent growth patterns and years of Saposhnikovia divaricata decoction pieces,which was applied to explore the effect of growth patterns and years on its quality. The appearance characteristic data of 55 batches of different growth patterns and years of S. divaricata were collected using PANTONE color card.High performance liquid chromatography( HPLC) was used to determine the contents of prim-O-glucosyl-cinmifugin,cimifugin,4-O-β-D-glucosyl-5-O-methylvisamminol and sec-O-glucosylhamaudol. The content of alcohol soluble extract and water-soluble extract were determined by hot-dip method. The content of volatile oil was determined by steam distillation. The correlation between growth patterns and years and the contents of 4 chromones,extracts and volatile oil were analyzed by modern statistical methods. Also,the method of comprehensively evaluating the quality of Chinese herbal pieces was developed by combining the growth patterns and years,appearance and chemical indexes. MTT assay was used to evaluate the effects on the survival rate of RAW264. 7 cells at four different concentrations of chromones and LPS was used to stimulate well-growing RAW264. 7 cells to establish an inflammatory model. The contents of NO and TNF-α in cell supernatant were detected by NO test kit and ELISA method. The contents of alcohol soluble extracts and water-soluble extracts in different growth patterns and years are: wild products<perennial cultivation<annual cultivation; the contents of volatile oil are: wild products>perennial cultivation>annual cultivation; the contents of four chromones are: wild products>perennial cultivation and annual cultivation. There was no significant difference between the sum of the two indexes in the Pharmacopoeia of perennial cultivation and wild products. 4 chromones showed no toxicity to RAW264. 7 cells at 5 mg·L-1. The release of NO and TNF-α was inhibited by 4 chromones and the anti-inflammatory effect of cimifugin was the best. In summary,there are obvious differences in appearance characteristics,internal quality and effects between different growth patterns and years. It showed that the wild products were superior to the perennial cultivation and the perennial cultivation was superior to the annual cultivation. In order to alleviate the shortage of wild S. divaricata resources,it is suggested that the Chinese Pharmacopoeia standard should increase the character of decoction pieces of perennial cultivation,and properly raise the limit requirement of the sum of the two indexes in the Chinese Pharmacopoeia to ensure the clinical demands and effect.
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Animaux , Souris , Apiaceae/croissance et développement , Chromatographie en phase liquide à haute performance , Médicaments issus de plantes chinoises/normes , Monoxyde d'azote/métabolisme , Huile essentielle/analyse , Facteur de nécrose tumorale alpha/métabolismeRésumé
The planting area of Chinese medicinal materials is an important basis for formulating policies such as production and poverty alleviation of Chinese medicinal materials and is determining the quantity of medicinal materials trade. Accurately mastering the information of the distribution,area and yield of Chinese medicinal materials cultivation is the basis of the adjustment of the planting structure of traditional Chinese medicine. It is now the largest planting place of Mongolian traditional Chinese medicinal materials in Naiman banner that is belonging to Tongliao city,Inner Mongolia. It is of great significance to obtain the planting area of Mongolian Chinese medicinal materials in Naiman banner in time and effectively for the development of subsequent industries. In this study,Saposhnikovia divaricata,a medicinal plant planted in Naiman banner,was selected as an example,and the fusion 2 m resolution ZY-3 remote sensing image was used as the data source. Based on the ground survey data,the sample data of each typical ground object were selected,and the spectral characteristic curves of different ground objects were obtained,and the S. divaricata spectral information was obtained. Using the filtering texture analysis method based on probability statistics,five kinds of texture image display results under different texture filtering were compared and analyzed,and finally the S. divaricata texture features based on information entropy are determined. The distribution range and planting area of S. divaricata in Naiman banner were extracted and interpreted by using the texture and spectral information of remote sensing images. The results showed that: S. divaricata was mainly distributed in the northeast and central south of Naiman banner,and the planting area was 5 336 mu( 1 mu≈667 m2). The field verification data were in good agreement with the remote sensing interpretation results,and the difference was small. It shows that the combination of spectral information and texture information can realize the discrimination of S. divaricata,and the interpretation results can provide a reference for the county to formulate the poverty alleviation action of Chinese medicinal material industry and the economic development plan of agricultural producing areas.
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Agriculture , Apiaceae , Chine , Médecine traditionnelle chinoise , Plantes médicinalesRésumé
Polysaccharide from traditional Chinese herb, Saposhnikovia divaricata (Turcz.) Schischk. (SD) was extracted, fractionated and characterized in this work. Four fractions were prepared. Their molecular weight, monosaccharide compositions, linkage modes and structural properties were characterized with SEC-MALS-RI, HPAEC-PAD, GC-MS and NMR. SDP1 was assigned as a 1, 4-α-glucan with small amount of O-6 linked branches. SDP2 contained a big amount of the 1, 4-α-glucan and a small amount of arabinogalactan, while SDP3 possessed relatively lower amount of the 1, 4-α-glucan and a big amount of the arabinogalactan. SDP4 was defined as a pectic arabinogalactan. Four fractions showed antioxidant activities in both molecular and cellular levels and their activity was ranked as SDP4 ≈ SDP3>SDP2>SDP1. The 1, 4-α-glucan in SDP1 had the weakest, while SDP3 and SDP4 showed similar and the highest antioxidant activity. The arabinogalactan was the major component of both SDP3 and SDP4, which significantly contributed to the antioxidant activity of SDP.
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Saposhnikovia divaricata is a valuable Chinese medicinal herb; the transformation from vegetative growth to reproductive growth may lead to the decrease of its pharmacological activities. Therefore, the study of bolting and flowering for Saposhnikovia divaricata is warranted. The present study aimed to reveal differentially expressed genes (DEGs) and regularity of expression during the bolting and flowering process, and the results of this study might provide a theoretical foundation for the suppression of early bolting for future research and practical application. Three sample groups, early flowering, flower bud differentiation, and late flowering (groups A, B, and C, respectively) were selected. Transcriptomic analysis identified 67, 010 annotated unigenes, among which 50, 165 were differentially expressed including 16, 108 in A vs B, and 17, 459 in B vs C, respectively. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway functional classification analysis were performed on these differentially expressed genes, and five important pathways were significantly impacted (P ≤ 0.01): plant circadian rhythm, other glycan degradation, oxidative phosphorylation, plant hormone signal transduction, and starch and sucrose metabolism. Plant hormone signal transduction might play an important role in the bolting and flowering process. The differentially expressed indole-3-acetic acid (IAA) gene showed significant down-regulation during bolting and flowering, while the transport inhibitor response 1 (TIR1) gene showed no significant change during the bolting process. The expression of flowering related genes FLC, LYF, and AP1 also showed a greater difference at different development stages. In conclusion, we speculate that the decrease in auxin concentration is not caused by the degrading effect of TIR1 but by an alternative mechanism.
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Apiaceae , Génétique , Fleurs , Génétique , Analyse de profil d'expression de gènes , Régulation de l'expression des gènes végétaux , Réseaux de régulation génique , Gènes de plante , ARN des plantes , Génétique , Reproductibilité des résultatsRésumé
AIM To study the chemical constituents of the secondary metabolites of endophytic fungi DL02 from Saposhnikovia divaricata (Turcz.) Schischk.and their biological activities.METHODS The ethyl acetate and methanol extract leavening from S.divaricata was isolated and purified by silica and Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The antifungal and antibacterial activities were determined by the minimum inhibitory concentration method.RESULTS Seven compounds were isolated and identified as β-sitosterol (1),kojic acid (2),stigmasta-7,22-diene-3β,5α,6α-triol (3),uracil (4),allantoin (5),erythritol (6) and adenosine (7).Compound 4 had strong inhibitory effects on pseudomonas aeruginosa with the MIC values of 31.3 μg/mL.CONCLUSION Compounds 2-5 and 7 are isolated from this fungus for the first time.Compounds 4-5 have strong biological activities.
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Saposhnikovia divaricata is a valuable Chinese medicinal herb; the transformation from vegetative growth to reproductive growth may lead to the decrease of its pharmacological activities. Therefore, the study of bolting and flowering for Saposhnikovia divaricata is warranted. The present study aimed to reveal differentially expressed genes (DEGs) and regularity of expression during the bolting and flowering process, and the results of this study might provide a theoretical foundation for the suppression of early bolting for future research and practical application. Three sample groups, early flowering, flower bud differentiation, and late flowering (groups A, B, and C, respectively) were selected. Transcriptomic analysis identified 67, 010 annotated unigenes, among which 50, 165 were differentially expressed including 16, 108 in A vs B, and 17, 459 in B vs C, respectively. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway functional classification analysis were performed on these differentially expressed genes, and five important pathways were significantly impacted (P ≤ 0.01): plant circadian rhythm, other glycan degradation, oxidative phosphorylation, plant hormone signal transduction, and starch and sucrose metabolism. Plant hormone signal transduction might play an important role in the bolting and flowering process. The differentially expressed indole-3-acetic acid (IAA) gene showed significant down-regulation during bolting and flowering, while the transport inhibitor response 1 (TIR1) gene showed no significant change during the bolting process. The expression of flowering related genes FLC, LYF, and AP1 also showed a greater difference at different development stages. In conclusion, we speculate that the decrease in auxin concentration is not caused by the degrading effect of TIR1 but by an alternative mechanism.
Sujets)
Apiaceae , Génétique , Fleurs , Génétique , Analyse de profil d'expression de gènes , Régulation de l'expression des gènes végétaux , Réseaux de régulation génique , Gènes de plante , ARN des plantes , Génétique , Reproductibilité des résultatsRésumé
BACKGROUND: 70-80% of the world’s population uses herbal therapy and drug preparations of traditional medicine for health and first aid treatment [1]. SDS is used for relieving fever and also used as an anti-inflammatory medicine.1-2 It is also used in the treatment of cancer and infectious disease in Eastern and Chinese medicine. It is important to study the pharmacological actions and do a genome study in order to use natural resources wisely and to access biological effects on the plant. AIM: The study of the herbal preparation Saposhnikovia Divaricata on joint inflammation MATERIALS AND METHODS: All the experimental procedures and protocols used in the study were reviewed and approved by the Bio-Medical Ethical Committee of Mongolian National University of Mongolia. A dry extract of SDS is root was prepared by the lyophilization method and used in the study. CIA pathology model were determined by the David D Brand (2005) method. Statistical Analysis: Statistical analysis of data was performed by SPSS 16.0 program and analyzed statistically using criteria of Student t test. RESULTS: As a result the treatment group of mice with the use of the drug Saposhnikovia divaricata and groups of mice were treated with sodium salicylate, the body weight of mice increased significantly, and histological signs of inflammation were pronounced. In the mice of the control group who did not receive treatment, body weight decreased significantly, and histological signs of inflammation of the joints were expressed significantly. CONCLUSION: Preparation of Saposhnikovia divaricata decreased inflammation of arthritis.
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ObjectiveTo investigate the effects of Saposhnikovia divaricata extract combined with arsenic trioxide (ATO) on the proliferation and apoptosis in chronic myelogenous leukemia K562 cells. MethodsSaposhnikovia divaricata extract was prepared.Cultured K562 cells were treated with different concentration of Saposhnikovia divaricataextract or/and ATO for 48h. Cell proliferation was determined using the MTT assay. Apoptosis and cell cycle were detected using flow cytometry.ResultsThe MTT assay showed that Saposhnikovia divaricata extract of 750,1 000,1 250,1 500 μg/ml had a significantly proliferation inhibitory effect compared with control group, the inhibitory rates were 23.29% ± 3.31%, 48.30% ± 2.50%, 79.62% ± 3.41% and 88.94% ± 0.06%, respectively (allP<0.05); Saposhnikovia divaricata extract of 500 μg/ml combined with ATO of 1.0, 0.5 μg/ml significantly increased inhibitor rates compared with ATO of 1.0, 0.5 μg/ml (64.99% ± 5.18%vs. 44.48% ± 3.31%,38.59% ± 3.88%vs.26.30% ± 5.03%; allP<0.05). FCM showed that Saposhnikovia divaricata extract of 500 μg/ml combined with ATO of 2.0, 1.0, 0.5 μg/ml significantly increased apoptotic rate compared with ATO group of 2.0, 1.0, 0.5 μg/ml (33.97% ± 0.59%vs.20.97% ± 2.17%, 13.53% ± 0.47%vs.9.77%±0.64%、6.63%±&0.40%vs.4.00%±0.46%; allP<0.05 ). Cell cycle results showed that Saposhnikovia divaricata extract of 500μg/ml combined with ATO of 2.0,1.0, 0.5μg/ml significantly increased the rate of S phase compared with ATO group of 2.0, 1.0, 0.5 μg/ml (60.25 ± 2.59%vs.55.61 ± 1.28%, 60.89 ± 1.53%vs.37.96 ± 1.02%, 47.76 ± 0.87%vs.39.90 ± 0.92%; allP<0.05).ConclusionsSaposhnikovia divaricataextract could obviously inhibit the proliferation of K562 cells and enhance the apoptotic effect of ATO. ATO could induce a G2/M phase arrest, while Saposhnikovia divaricata extract combined with ATO could induce a S phase arrest in K562 cells.
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Objective: To extract Saposhnikovia divaricata polysaccharides (SPS) and to analyze the characteristics of their physicochemistry and structures. Methods: Two main polysaccharides, SPS0 and SPS1, were obtained by DEAE-52 and Sephadex G-200 column chromatography; UV spectrophotometer, heat stability test, IR spectrometry, Congo-red test, environmental scanning electron microscope (ESEM), and atomic force microscope (AFM) were used to determine the physicochemical, morphological, and structural properties of polysaccharides. Results: The uronic acid contents of SPS0 and SPS1 were 12.07% and 0.95%, respectively. IR spectrometry showed SPS0 had a specific absorption peak of -COOH. Congo-red test indicated that only SPS0 had a triple helix conformation. ESEM and AFM showed that SPS0 was neat and orderly, with a hyper branched structure, but irregular figure and fewer branches were observed in SPS1. Conclusion: SPS0 is an acidic and hyper branched polysaccharide, while SPS1 is a homogeneous polysaccharide including β-D-pyranose sugar residues with better heat stability.
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Objective: To study the correlation between the content of volatile oil and the rhizosphere soil from the roots of Saposhnikovia divaricata collected from different regions at different harvest time. Methods: The volatile oil from the roots of S. divaricata was extracted by steam distillation and analyzed. According to the National Standard of the People's Republic of China, we determined the pH value, total nitrogen, hydrolysable nitrogen, available phosphorus, available potassium, and organic matter in the rhizosphere soil of S. divaricata. The obtained data were statistically analyzed with SPSS 13.0 software. Results: The content of volatile oil in the roots of S. divaricata presented a trend of decrease with different collection regions from south to north and the main chemical components were significantly different, in which 1-(1-formylethyl)-4-(1-buten-3-yl)-benzene and β-bisabolene were found as the principal components. There was a significantly negative power function relationship between the content of volatile oil and the total nitrogen, a significantly non-linear correlation between the content of volatile oil and the hydrolysable nitrogen, but a good correlation between content of volatile oil and available phosphorus, available potassium, and the content of organic matter was found only in the first sample. There were significant good correlations of the pH value and hydrolysable nitrogen with available phosphorus, hydrolysable nitrogen, and available phosphorus with available potassium, hydrolysable nitrogen, available phosphorus, and available potassium with organic matter in rhizosphere soil (P < 0.01). The significant correlation of hydrolysable nitrogen, available potassium with pH was found (P < 0.05). Conclusion: There is no significant difference among the contents of volatile oil extracted from S. divaricata collected at different harvest time. There is a negative correlation between the total nitrogen content in rhizosphere soil and the content of volatile oil, namely, the productivity of volatile oil decreases with the increase of total nitrogen content, while the optimal contents of hydrolysable nitrogen, available phosphorus, available potassium, and organic matter could accumulate the content of volatile oil. On the other hand, pH value may affect the content of volatile oil by modulating the other chemical properties of the rhizosphere soil.
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Objective: To develop an HPLC-DAD method for the simultaneous determination and comparative analysis on the contents of prim-O-glucosylcimifugin, 4′-O-β-D-glucosyl-5-O-methylvisamminol, cimifugin, sec-O-glucosylhamaudol, psoralen, imperatorin, bergapten, and xanthotoxin in the roots of Saposhnikovia divaricata from Longxi areas. Methods: The extracts were obtained by methanol reflux method and the contents of the eight compounds were determined by HPLC-DAD. The mobile phase, consisting of acetonitrile-water, was programmed for a gradient elution. The flow rate was 1.0 mL/min, the detection wavelength was 254 nm, and the column temperature was 25°C. Results: Excellent linearity with correlation coefficents (r) of 0.9992-0.9999 was obtained. The average recoveries of the eight compounds were 96.2%-104.1% and all RSD values were less than 3%. The contents of the four coumarins in the roots of S. divaricata were much less than those four chromones. The contents of the four coumarins in the roots of Carum carvi and Peucedahum ledebourielloides were more than those in S. divaricta, while no chromones were detected except sec-O-glucosylhamaudol in P. ledebourielloides. Conclusion: The method appears to be simple, accurate, and well reproducible, which could be used for the simultaneous determination of the above-mentioned eight compounds in S. divaricata. According to the above analysis, C. carvi and P. ledebourielloides could not be used as the succedaneum of S. divaricata on the basis of the eight compound contents.
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High-speed counter-current chromatographic ( HSCCC ) method was successfully used to separate and purify prim-O-glucosylcimifugin and 5-0-methylvisammioside from Saposhnikovia divaricata(Thicz. ) Schis-chk. ,a traditional Chinese herb,with a solvent system composed of ethyl acetate-n-butanol-water(2:7:9,V/V). The lower phase of the system was used as the mobile phase at the flow rate of 2.0 mL/min,and the upper phase was used as the stationary phase. The separation produced a total 13.9 mg prim-O-glucosylcimifugin and 25.0 mg 5-0-methylvisammioside with the purity of 98. 1% and 99.2% determined by high performance liquid chromatography (HPLC) from 316 mg of crude sample from S. divaricata. The structures of isolated compounds were identified by ESI-MS,~1H NMR and ~(13)C NMR.
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Object To investigate the reason of the low germination and sprout rates by studying on inhibitory activities of the crude extracts of the seed of the dried root of Saposhnikovia divaricata (Turcz.) Schischk (DRSD) on Brassica, wheat and the seed of DRSD. Methods The ether extract and residue of the crude extract were prepared and treated by paper chromatography. The inhibitory activities of different fractions with different Rf value were determined. Effect of steeping the seed in warm water at 41 ℃ and 45 ℃ for different periods of time was also studied, respectively. Results There are intrinsic inhibitor in the seed of DRSD with strong activity. The portion of its ether extract with Rf 0.6 showed the strongest inhibitory activity for the germination of Brassica seed. The crude extract of the seed of DRSD also has the stronger inhibitory activity to the germination and growth of tender root of S. divaricata and has the same effect on aerial part of wheat sprouts. Steeping with warm water and solvent can remove most of the intrinsic inhibitor. Conclusion Intrinsic inhibitor in the seed of DRSD is the main factor that results in the low germination and sprouting slowly after seeding.
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Objective: A reversed phase HPLC method was described for determination of prim O glucosylcimifugin and 4′ O ? D glucosyl 5 O methylvisamminol in Ganmaoqingre Granules. Methods:The sample was separated on ODS column with mobile phase of methanol 40 mmol?L -1 sodium acetate pH 6.9 (35∶65) for prim O glucosylcimifugin and H 2O methanol THF(62∶38∶1) for 4′ O ? glucosyl 5 O methylvisamminol. The flow rate was 0.8 mL?min -1 , and the detection was set at 254 nm. Results: The calibration curves were linear in the range of 0.72 ?g?mL -1 ～6.5?g?mL -1 for prim O glucosylcimifugin and 0.92?g?mL -1 ～16.5?g?mL -1 for 4′ O ? D glucosyl 5 O methylvisamminol( r =0.9999). The average recovery was 100.3% and 94.7%. The content of prim O glucosylcimifugin and 4′ O ? D glucosyl 5 O methylvisamminol in Ganmaoqingre Granules was 0.133 mg?g -1 and 0.167 mg?g -1 , respectively. Conclusion: The method is fast and specific for both constitutents of Ganmaoqingre Granules.