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OBJECTIVE To establish the methods for simultaneous determination of rutin， forsythiaside A， （+）-pinoresinol-4- O-β-D-glucopyranoside， forsythin and forsythigenin in Forsythia suspensa flower. METHODS UPLC method was adopted. The determination was performed on ACQUITY UPLC HSS T3 C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution （gradient elution） at the flow rate of 0.3 mL/min. The detection wavelengths were set at 275 nm （0-8 min），330 nm （8-10.5 min），275 nm （10.5-32 min）， respectively. The column temperature was 25 ℃， and sample size was 1 μL. Taking rutin as reference， the content of each component was determined by quantitative analysis of multi-components by single-marker （QAMS） method， and then compared with external standard method. RESULTS The contents of forsythiaside A， （+）-pinoresinol-4-O-β-D- glucopyranoside， forsythin and forsythigenin by QAMS were 7.472-7.671， 2.919-2.986， 1.439-1.486， 1.523-1.566 mg/g； the results obtained by the external standard method were 7.454-7.664， 2.913-2.996， 1.444-1.484， 1.519-1.562 mg/g， respectively. There was no significant difference in the measurement results between the two methods， with a relative deviation less than 1.0%. CONCLUSIONS This study successfully establishes the UPLC-QAMS method for simultaneous determination of five components in F. suspensa flower， and the results obtained by this method are not significantly different from those obtained by the external standard method. It can be used for quality control of F. suspensa flower.
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OBJECTIVE To investigate the effects of Forsythia suspensa ethanol extract on the proliferation， migration and invasion of lung cancer cells NCI-H226. METHODS As research objects， lung cancer cells NCI-H226 were divided into control group， F. suspensa ethanol extract low-， medium- and high-concentration groups （5， 10， 20 mg/mL）， activator group [10 mg/mL F. suspensa ethanol extract+0.5 μmol/L nuclear factor kappa B （NF-κB） signaling pathway activator PMA]， inhibitor group （10 mg/mL F. suspensa ethanol extract+10 μmol/L NF-κB signaling pathway inhibitor BAY 11-7082） and positive control group （20 μg/mL cisplatin）. Except for the control group of cells without intervention， all other groups of cells were cultured with corresponding drugs for 24 hours； the proliferation， migration and invasion of cells were all detected， and the proliferation rate， migration rate， and the number of invading cells were also calculated； protein expressions of NF-κB p65， NF-κB inhibitory protein α （IκBα）， phosphorylated NF-κB p65 （p-NF-κB p65） and phosphorylated IκBα （p-IκBα） were determined. RESULTS Compared with control group， the proliferation rate， migration rate， and the number of invading cells as well as the protein expressions of p- IκBα and p-NF-κB p65 were decreased significantly in F. suspensa ethanol extract groups and positive control group （P＜0.05）. Compared with F. suspensa ethanol extract medium-concentration group， the proliferation rate， migration rate， and the number of invading cells as well as above protein expressions were all decreased significantly in inhibitor group （P＜0.05）， while those of activator group were increased significantly （P＜0.05）. CONCLUSIONS F. suspensa ethanol extract can inhibit the proliferation， migration and invasion of lung cancer cells NCI-H226， and the mechanism of which may be related to the inhibition of NF-κB signaling pathway.
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OBJ ECTIVE To predict the quality marker （Q-Marker）of Forsythia suspensa . METHODS The fingerprints of 10 batches of F. suspensa were established by high performance liquid chromatography. The common peaks were confirmed. The candidate components of Q-Marker in F. suspensa were screened. The volatile oil of F. suspensa were analyzed by gas chromatography-mass spectrometry （GC-MS），and the candidate components of Q-Marker in volatile oil were screened. The network pharmacology analysis was performed for the candidate components of Q-Marker. The network diagram of the “candidate components of F. suspensa Q-Marker-target-pathway”was constructed to predict the Q-marker of F. suspensa . RESULTS & CONCLUSIONS Twenty-one common peaks were obtained for 10 batches of F. suspensa ，and four components were identified as phillyrin，forsythoside A ，pinoresinol-4-O-β-D-glucoside and rutin. Seven candidate components were obtained by GC-MS analysis，such as β-pinene，α-pinene，terpinen-4-ol，limonene，γ-terpinene，α-phellandrene，β-myrcene. By network pharmacology analysis， 16 key targets and 17 pathways were obtained. It was preliminarily predicted that phillyrin ， forsythoside A ， pinoresinol-4-O-β-D-glucoside，rutin，terpinen-4-ol，α-phellandrene，α-pinene and β-pinene were Q-marker of F. suspensa .
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Forsythiae Fructus is the dried fruit of Forsythia suspensa and the volatile compounds are its main bioactive components. According to the different harvest periods, F. suspensa can be divided into Qingqiao(mature F. suspensa) and Laoqiao(ripe F. suspensa). To investigate dynamic changes of volatile components in Qingqiao and Laoqiao samples collected at different periods, the present study extracted and analyzed the total volatile oils in Qingqiao and Laoqiao samples(four harvest periods for Qingqiao and two for Laoqiao) by steam distillation method. The results indicated that the content of volatile oils in F. suspensa samples at different harvest periods was significantly different. The content of volatile oils in Qingqiao samples(except those harvested in the first period) was higher than that of Laoqiao, and the content of volatile oils in both Qingqiao and Laoqiao increased with the harvest period. Furthermore, volatile compounds in F. suspensa were qualitatively analyzed by the gas chromatography-mass spectrometry(GC-MS), and 28 volatile compounds were identified. Chemometrics analyses including principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) were further applied to explore differential markers and dynamic changes of volatile components in Qingqiao and Laoqiao samples at different harvest periods. Finally, four volatile compounds, including α-pinene, sabinene, β-pinene, and 4-terpenol were selected as potential differential markers. The relative content of α-pinene and 4-terpenol was consistent with that of total volatile oils in the changing trend.
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Chemometrics , Forsythia , Fruit , Gas Chromatography-Mass Spectrometry , Oils, VolatileABSTRACT
OBJECTIVE To study the quality grade stand ard of the premature Forsythia suspensa . METHODS A total of 138 batches of premature F. suspensa were collected from the main producing areas of F. suspensa in China. According to 2020 edition of Chinese Pharmacopoeia ，the contents of impurities ，moisture，ethanol-soluble extract ，volatile oil ，forsythin and forsythoside A in the premature F. suspense were determined ，and the qualified samples were screened. AHP-PCA mixed weighting method was used to give comprehensive weight to the indicators （except for the limit of impurity ）. The comprehensive score of the samples was calculated. The suggestions on the quality grade division of premature F. suspensa were put forward according to cluster analysis of K-mean value. RESULTS & CONCLUSIONS The contents of impurities ，moisture，ethanol-soluble extract ，volatile oil ，forsythin and forsythoside A in the premature F. suspense were 0-7.80％，1.60％-8.18％，13.13％-61.60％，0.21％-3.47％，0.02％-2.15％ and 0.79％-14.04％，respectively；average contents of them were 1.24％，4.97％，34.88％，2.01％，0.42％，6.86％，respectively. Totally 47 batches of 138 batches were qualified in all indexes. It is suggested that the quality grade of the premature F. suspense can be divided into three grades ：in first grade of F. suspense ，the contents of volatile oil ，forsythin，forsythoside A ， ethanol-soluble extract and moisture were ≥2.40％，≥0.59％，≥8.34％，≥38.66％ and ≤4.99％，respectively；in second grade of F. suspense ，the contents of above indicators were ≥2.26％，≥0.41％，≥7.47％，≥32.58％ and ≤5.33％，respectively；in third grade of F. suspense ，the contents of above indicators were ≥2.15％，≥0.32％，≥4.60％，≥31.52％ and≤7.23％，respectively.
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OBJECTIVE：To establish the method for the content det ermination of 4 components in Forsythia suspensa flowers by drying in shade ，vacuum freeze-drying ，oven（30，50，70 ℃）and sun ，so as to evaluate the effects of different drying methods on the main components of F. suspensa flowers and screen the optimal drying method. METHODS ：UPLC method was adopted. The determination was performed on Waters ACQUITY UPLC BEH C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid aqueous solution （gradient elution ）at the flow rate of 0.3 mL/min. The detection wavelength was set at 230 nm， and column was 35 ℃. The sample size was 1 μL. Euclidean closeness（C）i of different drying methods was calculated by TOPSIS comprehensive analysis method ，and the optimal drying method was defined. RESULTS ：The linear range of forsythiaside A ， rutin，forsythin，（+）-pinoresinol-4-O-β-D-glucopyranoside were 0.007 5-0.037 7，0.027 4-0.137 2，0.001 9-0.009 5，0.005 6-0.028 8 µg（all r＞0.999）. RSDs of precision ，stability（32 h）and reproducibility tests were all lower than 2％. The recoveries were 97.27％-102.53％，100.53％-104.11％，98.45％-104.02％，98.66％-104.82％，respectively；and all RSDs ＜3％（n＝3）. The contents were 1.645 8-4.987 9，11.730 2-20.978 0，0.875 5-2.005 0，2.366 0-5.535 7 mg/g. The content of forsythiaside A was the highest after drying at 30 ℃，rutin and （+）-pinoresinol-4- O-β-D-glucopyranoside were the highest after vacuum freeze-drying，forsythiaside was the highest after drying at 50 ℃ . Results of TOPSIS analysis showed that Ci of F. suspensa flowers by drying in shade ，vacuum freeze-drying ，oven（30，50，70 ℃）and sun were 0.079 9，0.553 5，0.495 4， 0.503 8，0.157 9，0.217 2，respectively；the order of Ci was vacuum freeze-drying ＞ 50 ℃ oven drying ＞ 30 ℃ oven drying＞sun drying ＞70 ℃ oven drying ＞ shade drying. CONCLUSIONS：Established method is simple ，reproducible and can be used for the content determination of 4 components in F. suspensa flowers. The samples are preferably dried by vacuum freeze-drying，followed by 50 ℃ oven drying ，30 ℃ oven drying ， and then dried in the sun and oven at 70 ℃ and finally in the shade.
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OBJECTIVE：To e stablish and compare HPLC fingerprints of green Forsythia suspensa and grown F. suspensa ，and to conduct cluster analysis and principle component analysis. METHODS ：HPLC method was adopted. The determination was performed on Hypersil C 18 column with mobile phase consisted of acetonitrile- 0.1％ formic acid （gradient elution ）. The detection wavelength was 235 nm and column temperature was 25 ℃ with the flow rate of 1.0 mL/min. The sample size was 10 μL. HPLC fingerprints of 8 batches of green F. suspensa （Q1-Q8）and 6 batches of grown F. suspensa （L1-L6）were drawn ，with phillyrin as reference；the similarity evaluation was conducted by using Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition），and common peak was confirmed. Cluster analysis and principal component analysis were carried out with SPSS 23.0 software. RESULTS ：There were 19 common peaks for green F. suspensa and grown F. suspensa ，among which 6 peaks were identified，i.e. forsythoside A ，rutin，pinoresinol-β-D-glucoside，phillyrin，quercetin and phillygenin ；the similarities of HPLC fingerprints from green F. suspensa and grown F. suspensa were 0.351-0.767；results of cluster analysis showed that green F. suspensa and grown F. suspensa were classified into 4 categories，among which L 1-L6 were clustered into one category ，Q1 was clustered into one category ，Q2-Q6 were clustered into one category ；Q7-Q8 were clustered into one category. The results of principal component analysis showed that the cumulative variance contribution rate of the first three principal components was 83.14％， L1-L6 distribution was close ，Q2-Q6 distribution was close ，Q7-Q8 distribution was close ，and Q 1 distribution was independent ， which was consistent with the results of cluster analysis. CONCLUSIONS ：There were significant differences in the common peaks of fingerprint of green F. suspensa and grown F. suspensa of similarity eraluation ，cluster analysis and principle component analysis，the established HPLC fingerprint can be used for comprehensive evaluation and quality comparison of green F. suspensa and grown F. suspensa .
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Objective: To screen inhibitors targeting SARS-CoV-2 S protein-ACE2 interaction by molecular docking. Methods: Candidate natural products were collected from Selleck China natural product library (Catalog No. L1400, 2 054 natural products). The structure of SARS-CoV-2 S protein-ACE2 had been determined by Qiang Zhou team (PDB: 6M17). The molecular docking was performed by Discovery Studio. Results: Based on the virtual amino acid mutation experiment which determined the key amino acids, the binding cavity was created. Then, 11 compounds were screened out from the natural compound library: digitonin, Lonicera grisea saponin A, forsythiaside B, L. grisea saponin B, Dipsacus asperges saponin B, hederacoside D, platycodon D, echinacoside, ginsenoside Rb2, ginsenoside Rc, and chlorogenic acid C. Conclusion: The 11 potential inhibitors targeting SARS-CoV-2 S protein-ACE2 interaction were screened out from natural products library, which provides a reference for the research of new anti SARS-CoV-2 drugs.
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Objective: To compare the antidiarrheal effects of Mongolian medicine, Holarrhena antidysenterica, Forsythia suspensa and Cynanchum thesioides on diarrhea model rats and investigate its effects on serum DAO (diamine oxidase), cAMP (cyclic adenosine phosphate), TNF-α (tumor necrosis factor-α), ATPase and calcium ions. Methods: The normal control group, model group, H. antidysenterica low-dose and high-dose groups, F. suspensa low dose and high-dose groups, C. thesioides low dose and high dose groups were set. Except the normal control group, the other groups were ig administrated water decoction of Cassia angustifolia to establish diarrhea model; After the success of the model, the rats in treatment groups were administrated by gastric drug for 7 d, the type mental state, diarrhea and body weight changes were observed. the abdominal aortic blood was obtained at the last day of fasting 12 h after the administration. DAO, cAMP, TNF-α, ATP enzyme, OD value of the calcium ions in serum were determined by using ELISA (enzyme-linked immune detection reagent) kits. Results: General status: except for the normal group, the mental state of the rats in the other groups was depressed after modeling, the fur color of them was significantly decreased and the body weight was decreased. The diarrhea rate was 100% on the 4th day after modeling. Compared with the model group, there were significant differences in the number of loose stools, grade of loose stools and diarrhea index (P < 0.05, 0.01) in each administration group. The serum DAO, TNF-α, ATPase, cAMP and calcium ion OD values were compared: the serum concentrations of DAO in H. antidysenterica low dose group, C. thesioides low dose group and F. suspensa high and low dose groups of rats were significantly lower than model group with significant differences (P < 0.05), and were significantly higher than normal group. The serum concentration of TNF-α in C. thesioides high and low dose groups were lower compared with model group (P < 0.05). The serum ATPase in C. thesioides high and low dose groups had significant difference (P < 0.05) compared with model group. The serum concentration of cAMP in H. antidysenterica high-dose group and F. suspensa low-dose group was significantly lower compared with model group with significant differences (P < 0.05). Serum Ca2+ concentration in the drug administration groups was significantly different from that in the model group (P < 0.05). Conclusion: The antidiarrheal effect of C. thesioides is better than that of H. antidysenterica and F. suspensa.
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Objective: To investigate the genetic diversity of Forsythia suspensa from different populations. Methods: ISSR markers were used to analyze the genetic diversity of 25 populations of F. suspensa. Percentage of species level polymorphic bands (P), Nei’s genetic diversity index (H), and Shannon’s information index (I) of genetic information were calculated by POPGEN 32. UPGMA relationship dendrogram was clustered by NTSYS. Results: Thirteen primers produced 353 bands and P was 100%. H and I were 0.252 3 and 0.394 0, and genetic differentiation coefficient (Gst) and gene flow (Nm) were 0.331 8 and 1.007 0 within the population levels. The genetic distance varied from 0.031 0 to 0.155 5. NTSYSpc software was used to cluster the system. A total of 25 clusters were divided into two categories and seven groups. Cluster analysis was conducted among Forsythia individuals. A total of 195 samples were divided into two categories and five groups. Conclusion: The genetic diversity among the populations of F. suspensa is at higher level. However, the genetic distance among the Forsythia population is not related to the geographical distance, which is mainly dependent on the ecological factors and the growth environment.
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OBJECTIVE： To establish a method for simultaneous determination of α-pinene， β-pinene， limonene and α-terpineol in volatile oil of Forsythia suspensa. METHODS： GC method was adopted. The determination was performed on HP-5 capillary column through temperature-programmed route. The inlet temperature was 230 ℃， and detector temperature was 250 ℃； split sampling was applied （split ratio of 8 ∶ 1）； the air flow rate was 300 mL/min， the hydrogen flow rate was 30 mL/min， the tail gas flow rate was 30 mL/min， and the injection volume was 1 μL. Using limonene as internal reference， relative correction factors of α-pinene， β-pinene and α-terpineol were established， and the reproducibility of relative correction factors were investigated by using different chromatographs and columns， and chromatographic peak location of components was measured. The contents of above components were calculated with QAMS， and then compared with the results of external standard method. RESULTS： The linear range of α-pinene， β-pinene， limonene and α-terpineol were 16.5-990.0， 38.1-2 287.5， 8.2-491.2， 2.4-142.5 μg/mL， respectively （r≥0.999 1）. RSDs of precision， reproducibility and stability tests were all lower than 3％ （n＝6）. Average recoveries were 99.7％-105.5％（RSD＜4％，n＝9）. Compared with limonene （1.00），the average relative correction factors of α-pinene， β-pinene and α-terpineol were 0.91，0.86 and 1.11（n＝3）； relative retention time were 0.69-0.74， 0.81-0.86， 1.25-1.35（RSD＜3％，n＝3）. By using different chromatographs and columns， RSDs of relative correction factors were 0.21％-4.65％（n＝6）. Compared with external standard method， determination results of above 4 components were consistent （the absolute value of relative error were all less than 7％）. CONCLUSIONS： QAMS can be used for simultaneous determination of 4 kinds of effective components in volatile oil from F. suspensa.
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OBJECTIVE： To establish the method for content determination of volatile oil of Schizonepeta tenuifolia and Forsythia suspensa， and to optimize the extraction technology of the volatile oil. METHODS： The contents of β-pinene and pulegone were determined by GC method. The determination was performed on Hp-5 capillary column. The detector was hydrogen flame ion detector with programmed temperature. The sample size was 0.5 μL， the split ratio was 70 ∶ 1， the carrier gas was nitrogen， the inlet temperature was 250 ℃， the detector temperature was 280 ℃， the air flow rate was 390 mL/min， the hydrogen flow rate was 36 mL/min， the tail flow rate was 15 mL/min， and the nitrogen flow rate was 1 mL/min. Based on single factor test， orthogonal test combined with information entropy method were used to optimize the extraction technology of S. tenuifolia and F. suspensa using soaking time， extraction time， material-liquid ratio and forsythia grain size as factors， with the extraction amount of volatile oil， the content of β-pinene and pulegone and their comprehensive score as indexes. RESULTS： The linear range of β-pinene and pulegone 1.575-7.875（r＝0.999 9） and 1.892-9.46 μg（r＝0.999 7）， respectively. The limits of quantitation were 0.10 and 0.25 μg； the limits of detection were 0.03 and 0.08 μg； RSDs of precision， stability and reproducibility tests were less than 2％ （n＝6）； the recoveries were 97.77％-100.01％ （RSD＝0.93％，n＝9） and 96.47％-99.00％（RSD＝0.89％， n＝9）. The optimal extraction technology was soaking 2 h， extracting for 6 h， 10-fold water （mL/g）， half a clove of granularity. Under this condition， the extraction amount of volatile oil， the contents of β-pinene and pulegone were 3.6 mL， 1 450.4 mg， 127.6 mg， respectively. RSD were 1.62％， 0.20％， 1.42％. CONCLUSIONS： Established method is simple， accurate and reproducible， and the optimal extraction technology is stable and feasible.
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Forsythia suspensa is a herbal medicine that widely used for heat-clearing and detoxification in clinical practice. However, the molecular mechanism of its heat-clearing and detoxifying effect is still unclear. Based on the theory and methods of network pharmacology, the efficacy of the heat-clearing and detoxification of Forsythia suspensa was analyzed in this study. A total of 114 of compounds in Forsythia suspensa were collected, and 15 of effective compounds were obtained by analyzing the bioavailability (OB) and drug-like properties (DL). Then 26 corresponding targets were obtained using reverse pharmacophore-docking method. Using the BioGPS database, the organ location of the target initially was revealed. The compound-targetdisease network model of Forsythia suspensa was constructed by using the Cytoscape, which showed that the material basis of the heat-clearing and detoxification of Forsythia suspensa was to synthesize and synergize the effects by combining various active ingredients of multiple targets, simultaneously. This study explains the scientific mechanism of the heat-clearing and detoxification of Forsythia suspensa, and provides a theoretical foundation for clinical rational usage of Forsythia suspensa.
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To investigate the feasibility of vapor permeation membrane technology in separating essential oil from oil-water extract by taking the Forsythia suspensa as an example. The polydimethylsiloxane/polyvinylidene fluoride (PDMS/PVDF) composite flat membrane and a polyvinylidene fluoride (PVDF) flat membrane was collected as the membrane material respectively. Two kinds of membrane osmotic liquids were collected by self-made vapor permeation device. The yield of essential oil separated and enriched from two kinds of membrane materials was calculated, and the microscopic changes of membrane materials were analyzed and compared. Meanwhile, gas chromatography-mass spectrometry (GC-MS) was used to compare and analyze the differences in chemical compositions of essential oil between traditional steam distillation, PVDF membrane enriched method and PDMS/PVDF membrane enriched method. The results showed that the yield of essential oil enriched by PVDF membrane was significantly higher than that of PDMS/PVDF membrane, and the GC-MS spectrum showed that the content of main compositions was higher than that of PDMS/PVDF membrane; The GC-MS spectra showed that the components of essential oil enriched by PVDF membrane were basically the same as those obtained by traditional steam distillation. The above results showed that vapor permeation membrane separation technology shall be feasible for the separation of Forsythia essential oil-bearing water body, and PVDF membrane was more suitable for separation and enrichment of Forsythia essential oil than PDMS/PVDF membrane.
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Objective To study the chemical constituents from the fruits of Forsythia suspensa. Methods Compounds were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, Sephadex LH-20 and reversed-phase HPLC. Their structures were elucidated by physiochemical properties and spectral analysis. Results Two new compounds from the 50% ethanol extract of F. suspensa were isolated and identified as (2R,3S)-3-(4-hydroxy-3-methoxyphenyl) - 3-methoxypropane-1,2-diol (1) and 2-methylhexane-1,2,3,6-tetraol (2). Conclusion Compound 1 is a new epimer of a known phenylpropanoid named (2R,3S)-forsythiayanoside D, and compound 2 is a new compound named rengyquaol.
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Objective: To study the chemical constituents from the fruits of Forsythia suspensa. Methods: Compounds were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by physiochemical property and spectral analysis. Results: Thirteen compounds were isolated and identified as salidroside (1), forsythoside E (2), quinolacetic acid (3), rengyolone (4), senecio lactone (5), cleroindicin C (6), 4-hydroxy-4-isopropylcyclohex-1-enecarboxylic acid (7), oleuropeic acid (8), rel-(1S,2S,4S)-trihydroxy-p-menthane (9), azelaic acid (10), 2-hydroxy-succinic acid 4-methylester (11), glochidioboside (12), and (+)-isolariciresino-9-O-β-D-glucopyranoside (13). Conclusion: Compounds 5-13 are isolated from genus Forsythia Vahl. for the first time.
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OBJECTIVE: To study the chemical constituents from the fruits of Forsythia suspensa (Thunb.) Vahl.. METHODS: Compounds were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, Sephadex LH-20 and reversed-phase HPLC. Their structures were elucidated by physiochemical properties and spectral analysis. RESULTS: Sixteen compounds were isolated and identified as 3, 4, α-trihydroxy-methyl phenylpropionate (1), protocatechaldehyde (2), vanillic acid (3), p-hydroxybenzoic acid (4), p-hydroxylbenzylalcohol (5), p-hydroxyphenylethyl alcohol (6), 2-(4-methoxyphenyl) acetaldehyde (7), 2-(4-hydroxyphenyl) acetic acid (8), 4-hydroxyphenethyl 2-(4-hydroxyphenyl) acetate (9), p-coumatic acid (10), methyl 2-(4-hydroxyphenyl) acetate (11), 3, 4-dihydroxyphenylethanol (12), 1, 2, 4-benzentriol (13), 1-(4- hydroxyphenyl)-2, 3 -dihydroxypropan-1-one(14), 4-hydroxybenzaldehyde (15), and isovanillic acid (16). CONCLUSION: Compounds 1-3, 5, 7, 9, 11, and 14 are isolated from this plant for the first time.
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Objective: RAPD technique has been used to construct DNA fingerprints and analyze the genetic diversity in 14 batches of Forsythia suspensa from different origins. Methods: Primers with good polymorphism were screened from 45 RAPD primers combinations by PCR, and the DNA fingerprints for 14 batches of F. suspensa were constructed by RAPD technique. Results: Eleven RAPD primers with good polymorphism were screened from 45 RAPD primers by PCR amplification, and 80 bands were amplified by the 11 RAPD primers with average 7.27 bands for each primer, of which 67 were polymorphic and PPB was 83.8%. The genetic similarity (GS) ranged from 0.4875 to 0.9625 in 14 batches of F. suspensa from different origins, which showed they had rich genetic diversity. The result of cluster analysis showed that F. suspensa was correlated with sample origin and was consistent with species distribution. Conclusion: The DNA fingerprints for 14 batches of F. suspensa are established based on the RAPD amplified bands.
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Objective: To study the chemical constituents from the fruits of Forsythia suspensa. Methods: Compounds were isolated by combination of various chromatographic techniques including column chromatography over macroporous resin, silica gel, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by physicochemical property and spectroscopic analysis. Results: Two compounds were isolated from the 50% ethanol extract of the fruits of F. suspensa and identified as 7R-suspensaside methyl ether (1) and 7S-suspensaside methyl ether (2). Conclusion: Compound 1 is a new compound named forsythoside K.
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OBJECTIVE:To study and compare the volatile constituents in Forsythia suspensa and old F. suspensa. METH-ODS:Steam distillation method was adopted to extract volatile oil. GC-MS was adopted to determine the content of volatile oil:the chromatographic conditions:the column was HP-5MS,inlet temperature was 200 ℃,the carrier gas was nitrogen,split ratio was 20 to 1,by programmed temperature,flow rate was 1 ml/min,the injection volume was 1 μl;MS conditions:ion source was electric bombardment ionization source,electron energy was 70 eV,the interface temperature was 280 ℃,ion source temperature was 230 ℃,quadrupole temperature was 150 ℃,electron multiplier voltage was 2 300 V,and mass scanning range was 29-350 amu. RESULTS:The extraction yield order of volatile oil was that F. suspensa heart was higher than F. suspensa,followed by F. suspensa shell and old F. suspensa;there were 18 common peaks in the volatile oil,and the content of β-pinene was the maxi-mum,followed by α-pinene,β-phellandrene,terpinen-4-ol and terpinene;in addition,4-methylene-1-cyclohexanone,3-cyclohex-ene-1-methanol,cumene formaldehyde,1,7-trimethyl-4-methylenedecahydro,palmitic acid,nutmeg aldehyde can only be detected in the volatile oil of old F. suspensa;while α-phellandrene,ocimene,α-terpineol,α-pinocarvone,cis-piperitol,bornyl acetate,α-cubebene and germacrene D can only present in F. suspensa. CONCLUSIONS:The main chemical components in the volatile oil of F. suspensa and old F. suspensa are basically the same,but there are also some differences. The discovery can provide some ba-sis for the material basic research in the pharmacodynamic differences of F. suspensa.