Forsythiaside B inhibits cerebral ischemia/reperfusion-induced oxidative stress injury in mice via the AMPK/DAF-16/FOXO3 pathway / 南方医科大学学报

OBJECTIVE@#To study the protective effect of forsythiaside B (FB) against cerebral oxidative stress injury induced by cerebral ischemia/reperfusion (I/R) in mice and explore the underlying mechanism.@*METHODS@#Ninety C57BL/6 mice were randomized into sham-operated group, middle cerebral artery occlusion (MCAO) model group, and low-, medium and highdose (10, 20, and 40 mg/kg, respectively) FB groups. The expression levels of MDA, ROS, PCO, 8-OHdG, SOD, GSTα4, CAT and GPx in the brain tissue of the mice were detected using commercial kits, and those of AMPK, P-AMPK, DAF-16, FOXO3 and P-FOXO3 were detected with Western blotting. Compound C (CC), an AMPK inhibitor, was used to verify the role of the AMPK pathway in mediating the therapeutic effect of FB. In another 36 C57BL/6 mice randomized into 4 sham-operated group, MCAO model group, FB (40 mg/kg) treatment group, FB+CC (10 mg/kg) treatment group, TTC staining was used to examine the volume of cerebral infarcts, and the levels of ROS and SOD in the brain were detected; the changes in the protein expressions of AMPK, P-AMPK, DAF-16, FOXO3 and P-FOXO3 in the brain tissue were detected using Western blotting.@*RESULTS@#In mice with cerebral IR injury, treatment with FB significantly reduced the levels of ROS, MDA, PCO and 8-OHdG, increased the activities of antioxidant enzymes SOD, GSTα4, CAT and GPx, and enhanced phosphorylation of AMPK and FOXO3 and DAF-16 protein expression in the brain tissue (P < 0.01). Compared with FB treatment alone, the combined treatment with FB and CC significantly reduced phosphorylation of AMPK and FOXO3, lowered expression of DAF-16 and SOD activity, and increased cerebral infarction volume and ROS level in the brain tissue of the mice (P < 0.01).@*CONCLUSION@#FB inhibits oxidative stress injury caused by cerebral I/R in mice possibly by enhancing AMPK phosphorylation, promoting the downstream DAF-16 protein expression and FOXO3 phosphorylation, increasing the expression of antioxidant enzymes, and reducing ROS level in the brain tissue.

Protective effect of forsythiaside A on acute lung injury in septic rats / 中华危重病急救医学

Objective:To observe the protective effect of forsythiaside A on acute lung injury (ALI) in septic rats.Methods:Male Sprague-Dawley (SD) rats were randomly divided into normal control group, sham operation group, sepsis model group, and forsythiaside A intervention group, with 10 rats in each group. The rats in the normal control group did not receive any intervention; the rats in the sham operation group only underwent abdominal surgery; and those in the model group and forsythiaside A intervention group received cecal ligation and puncture (CLP) to establish the sepsis rat model. The rats in the forsythiaside A intervention group were given 75 mL/kg of forsythiaside A within 0.5 hour after operation, and repeated after 6 hours. The rats in the sham operation group and model group were given the same amount of normal saline at the same time points. The lung tissues were collected for pathological examination 12 hours after operation. The lung homogenate was prepared, and enzyme-linked immunosorbent assay (ELISA) was used to detect tumor necrosis factor-α (TNF-α), interleukins (IL-1β, IL-6). The activity of superoxide dismutase (SOD) was detected by xanthine oxidase method, and the content of malonaldehyde (MDA) was detected by colorimetry. The expression of nuclear factor-κB p65 (NF-κB p65) was detected by Western blotting.Results:There was no significant pathological change of lung tissue in both normal control group and sham operation group, and there was no significant difference in each parameter between the two groups. The rats in the model group had interstitial infiltration of inflammatory cells, alveolar structure destruction, alveolar septum thicken, extensive alveolar hemorrhage, telangiectasia; the levels of TNF-α, IL-1β, IL-6, MDA and NF-κB p65 protein expression in lung tissue were significantly higher than those in the normal control group and sham operation group [TNF-α (ng/L): 132.81±16.15 vs. 45.08±5.98, 46.10±6.72, IL-1β (ng/L): 137.32±15.22 vs. 51.03±7.89, 50.92±8.13; IL-6 (ng/L): 138.39±14.28 vs. 51.68±7.03, 52.48±7.36; MDA (kU/g): 1.79±0.13 vs. 0.96±0.05, 0.97±0.05; NF-κB p65 protein (NF-κB p65/GAPDH): 2.82±0.23 vs. 1.76±0.12, 1.82±0.13; all P < 0.05], the activity of SOD decreased significantly (kU/g: 45.90±5.46 vs. 92.11±10.13, 93.36±10.56, both P < 0.05). The changes in lung histopathology in the forsythiaside A intervention group were obviously improved as compared with the model group, which showed less inflammatory cell infiltration, less alveolar septum thickening, less bleeding and more intact structures; the levels of TNF-α, IL-1β, IL-6, MDA and the expression of NF-κB p65 protein in lung tissue were significantly lower than those in the model group [TNF-α (ng/L): 72.48±9.78 vs. 132.81±16.15, IL-1β (ng/L): 83.85±12.46 vs. 137.32±15.22, IL-6 (ng/L): 81.88±11.89 vs. 138.39±14.28, MDA (kU/L): 1.29±0.09 vs. 1.79±0.13, NF-κB p65 protein (NF-κB p65/GAPDH): 2.29±0.19 vs. 2.82±0.23, all P < 0.05], SOD activity increased significantly (kU/g: 66.03±7.98 vs. 45.90±5.46, P < 0.05). Conclusions:Forsythiaside A can effectively alleviate ALI in septic rats. The mechanism may be related to down-regulate the expression of NF-κB p65 and reduce the level of inflammatory factors and free radicals in lung tissue, thereby against acute lung injury in septic rats.

Preparation and in vitro evaluation of forsythoside A-loaded exosomes / 中国中药杂志

A drug delivery system of forsythoside A-loaded exosomes(FTA-Exos) with high biocompatibility and low immunogenicity was established to investigate its impact on the migration of human lung epithelial adenocarcinoma A549 cells. The exosomes from A549 cells were extracted and purified by ultra-high speed centrifugation and ultrafiltration. FTA-Exos were prepared by ultrasonic incubation, and characterized by particle size analysis, transmission electron microscopy, and Western blot assay. The uptake of FTA-Exos by A549 cells was observed under the laser confocal microscope, and the impact of FTA-Exos on the migration of A549 cells was investigated by cell scratch assay. The results showed that the average particle size of the prepared FTA-Exos was(138.90±2.37) nm, which increased slightly after drug loading. The PDI was 0.291±0.013, and the average potential was(-10.1±0.66) mV. The FTA-Exos were spheroidal in appearance as observed by transmission electron microscope, with an obvious saucer-like double-layer membrane. Western blot assay indicated that the specific proteins CD63 and Alix were both expressed in exosomes. The laser confocal microscopy suggested that FTA-Exos were taken up by A549 cells and stably maintained in the cell for 4-8 h, and the fluorescence was significantly enhanced at 4 h. The scratch assay showed that the inhibitory effect of FTA-Exos on the migration of A549 cells was significantly stronger than that of forsythoside A(P < 0.05). In conclusion, the drug delivery system of FTA-Exos established in this study had good stability, reliable preparation process, and potent inhibitory effect on the migration of A549 cells in vitro, which can provide an important reference for subsequent in-depth research and application.

Effect of Forsythionin A on Gastrointestinal Motility Disorders in Mice Treated with Chemotherapy / 中国实验方剂学杂志

Objective:To observe the effect of forsythiaside A on gastrointestinal motility disorder induced by chemotherapy in mice， and explore the mechanism of forsythiaside A regulating gastrointestinal motility. Method:The 60 KM mice were randomly divided into normal group， model group， metoclopramide group （5 mg·kg-1） and forsythiaside A low， medium and high-dose groups （30， 60， 120 mg·kg-1）， 10 for each group， which include half male and half female. The above dose was given once a day for 4 consecutive days， which the intragastric volume was 10 mL·kg-1. One hour after 1rd day administration， equal volume of saline was intraperitoneally injected to the normal group， 2 mg·kg-1 cisplatin was intraperitoneally injected to the other groups with daily for 4 consecutive days. Observing the effects of forsythiaside A on gastric emptying and small intestinal propulsion on mice models， serum gastrin （GAS） and somatostatin （SS）， motilin （MTL）， vasoactive intestinal peptide （VIP） levels were examined by enzyme-linked immunosorbent assay （ELISA）. Activities of acetylcholinesterase （AChE） and total nitric oxide synthase （tNOS） in gastric antrum and ileum were detected by ELISA. The expression of AChE and inducible nitric oxide synthase （iNOS） in gastric antrum and ileum were detected by Western blot. Result:Compared with normal group， the gastric retention rate and small intestinal propulsion rate of the model group were significantly increased （P<0.01）， serum levels of MTL， GAS， SS and VIP， the AChE activity in the homogenate of ileum in the model group were significantly reduced （P<0.05，P<0.01）， while the tNOS activities in gastric antrum and ileum were significantly increased （P<0.05，P<0.01）. Protein expression of AChE in gastric antrum and ileum were significantly decreased （P<0.05）， and the expression level of iNOS protein was significantly increased in the model group （P<0.05）. Compared with model group， different doses of forsythiaside A can reduce the gastric residual rate and small intestinal propulsion rate of mice to varying degrees. Meanwhile forsythiaside A can increase the serum levels of MTL， GAS， SS， and VIP， and the AChE activity and protein expression levels in gastric antrum and ileum tissues were also increased， while tNOS activity and iNOS protein expression were decreased in gastric antrum and ileum （P<0.05，P<0.01）. Conclusion:Forsythiaside A can significantly ameliorate the delayed gastric emptying and small intestine hyperfunction induced by cisplatin in mice. Its mechanism to ameliorate gastrointestinal dysfunction caused by chemotherapy is related to the regulation of gastrointestinal AChE and NOS activity in gastric antrum and ileum and the regulation of gastrointestinal hormone levels.

Forsythiaside B inhibits the proliferation activity of HeLa cells though transcription factor NF-κB / 中国临床药理学与治疗学

AIM: To study the effects of forsythiaside B on HeLa cells proliferation by inhibiting NF-κB through p21/Cyclin E/CDK2 signaling pathway. METHODS: The cell activity of HeLa cells was observed by MTT assay after 24 h, 48 h and 72 h treatment with different concentration (1, 2, 4 μmol/L) of forsythiaside B. The clone formation of HeLa cells was evaluated with the condition of different concentrations of forsythiaside B. The expression of transcription factor NF-κB (p-p65 and p65) was determined, and the expression of p21, Cyclin E and CDK2 were detected as well.RESULTS:Forsythioside B inhibited the clone formation of HeLa cells in concentration dependent manner, and the inhibitory effect on HeLa cells' proliferation by forsythioside B was in both time and concentration dependent manner. Forsythioside B up-regulated p21 protein expression and down-regulated p-p65, Cyclin E and CDK2 protein expression. CONCLUSION:Forsythioside B inhibits the transcription factor NF-κB and affects the p21/Cyclin E/CDK2 signaling pathway, thus inhibiting the proliferation of HeLa cells.

Network pharmacology study of Yinqiao Jiedu Soft Capsules in treatment of COVID-19 / 中草药

Objective: To explore the mechanism of Yinqiao Jiedu Soft Capsules in the treatment of coronavirus disease 2019 (COVID-19). Methods: The interactions between 1 418 compounds of Yinqiao Jiedu Soft Capsules and 48 inflammatory target proteins related to COVID-19 were analyzed by molecule docking. The drug-target network was established to clarify the active compounds and potential targets. Results: The network analysis suggested 50 active compounds of Yinqiao Jiedu Soft Capsules, which were mainly flavonoids and triterpenoids, and 37 potential targets, mainly including MTOR, JAK3, ACE, ACE2, PIK3CA, TNF, AKT2, and MAP2K1. The results of molecular docking exhibited that forsythiaside and vitexin 2″-O-rhamnoside had good affinity with SARS-CoV-2 3CL hydrolase, and glycyrrhizic acid had good affinity with ACE2. Conclusion: The molecular mechanism of Yinqiao Jiedu Soft Capsules for COVID-19 may be involved in interfering SARS-CoV-2 replication and regulating the expression of inflammatory signaling pathway and the secretion of inflammatory cytokines.

Study on Changes of Extraction Rates of Forsythiaside A and Forsythin in Forsythia suspense Compatible with Other Medicinal Materials of Menshi Huwei Formula before and after Decoction / 中国药房

OBJECTIVE： To investigate the changes of extraction rates of forsythiaside A and forsythin in Forsythia suspensa compatible with other medicinal material of Menshi huwei formula before and after decoction. METHODS： HPLC method was used to determine the extraction amounts and to calculate the extraction rates of forsythiaside A and forsythin in F. suspensa （5 g×7 doses）， F. suspensa （5 g×7 doses） compatible with Pinelliae rhizoma praeparatum cum zingibere et alumine （PRZA）， Menshi huwei formula [including 6 ingredients as F. suspense （5 g×7 doses）， PRZA] after decocted with water. The determination was performed on Diamonsil C18 column with mobile phase consisted of acetonitrile-0.2％ formic acid solution （gradient elution）. The detection wavelength was set at 278 nm， and the column temperature was 30 ℃. The flow rate was 1.0 mL/min. RESULTS： The linear range of forsythiaside A and forsythin were 0.61-6.1， 0.246-2.46 μg （r＝0.999 7， 0.999 9）， respectively； RSDs of precision， stability （within 20 h） and reproducibility tests were all lower than 2％ （n＝6）. Average recovery rates were 96.10％-99.37％ （RSD≤2.36％，n＝6） respectively. In F. suspensa， extraction rates of forsythiaside A and forsythin were 96.90％ and 66.67％. In F. suspensa compatible with PRZA， extraction rates of them were 101.61％ and 54.55％. In Menshi huwei formula， extraction rates of them were 98.39％ and 84.85％. CONCLUSIONS： After F. suspensa is compatible with PRZA， the extraction rates of forsythiaside A is increased while forsythin is decreased. After compatible with other medicinal material in Menshi huwei formula， extraction rates of both are increased slightly.

Simultaneous Determination of 7 Components in Tanreqing Capsules by HPLC / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of 7 components in Tanreqing capsules.METHODS:HPLC method was adopted.The determination was performed on Ultimate XB C18 column with mobile phase consisted of acetonitrile-0.2％ formic acid (gradient elution) at a flow rate of 0.8 mL/min.The detection wavelength was set at 325 nm,and column temperature was 35 ℃.The sample size was 10 μL.RESULTS:The linear ranges of chlorogenic acid,isoforsythiaside A,forsythiaside A,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C and baicalin were 0.025-0.5 μg(r=0.999 6),0.025-0.5 μg (r=0.999 7),0.050-1.0 μg (r=0.999 9),0.025-0.5 μg((r=0.999 7),0.025-0.5 μg ((r=0.999 6),0.025-0.5 μg (r=0.999 6),0.750-1.5 μg(r=0.999 9),respectively.The limit of quantitation was no more than 1.5 ng,and the limit of detection was 0.5 ng.RSDs of precision,stability and reproducibility tests were all lower than 3.0％.The recoveries were 95.28％-106.30％ (RSD ranged 0.97％-2.14％,n=9).CONCLUSIONS:The method is simple,precise,stable and reproducible,and can be used for simultaneous determination of 7 components in Tanreqing capsules.

Simultaneous quantification of eight constituents in the seed of Forsythia suspense (Thunb.) Vahl by HPLC-MS/MS method / 中草药

Objective: To develop an HPLC-MS/MS method for the simultaneous determination of forsythoside A, phillyrin, forsythiaside, rutin, quercetin, isoquercitrin, ferulic acid, and hesperidin in the seed of Forsythia suspense (Thunb.) Vahl. Methods: Chromatographic separation was performed on a Diamonsil C18 column (150 mm × 4.6 mm, 5 μm) with linear gradient elution of methanol and 0.1% formic acid at a flow rate of 800 μL/min, and the injection volume was 10 μL. Multiple-reaction monitoring (MRM) was employed with switching electrospray ion source polarity in negative mode. The ion spray voltage was set at-4 500 V and the turbo spray temperature was maintained at 650℃. Results: All calibration curves showed good linearity within the test ranges. The average recoveries of the compounds ranged from 98.1% to 101.8%. The precisions (RSD) for the investigated components were less than 0.84%. The contents of forsythoside A, phillyrin, forsythiaside and rutin are higher than quercetin, isoquercitrin, ferulic acid and hesperidin. And the average contents of forsythoside A, phillyrin, forsythiaside and rutin are 1 609.78, 80.08, 86.64 and 373.86 μg/g, respectively. Conclusion: An efficient, rapid, and sensitive method is first established for the qualitation and quantification of eight major components in the seed of Forsythia suspense (Thunb.) Vahl. The satisfactory results demonstrate that the method could be applied as a reliable quality control method for the seed of Forsythia suspense (Thunb.) Vahl.

Determination of seven components in Pifubingxuedu tablet by HPLC method / 药学实践杂志

Objective To establish a HPLC method for simultaneous determination of seven constituents in Pifubingxuedu tablet.Methods The determination was performed on Kromasil-C18 column (4.6 mm × 250 mm, 5 μm).The mobile phase was acetonitrile-0.5％ phosphoric acid aqueous solution with a flow rate of 0.8ml/min.The detector was PDA and detection wavelength was set at 245,325,403 nm, respectively.Results A method has been established for the determination of chlorogenic acid, hydroxy Safflower Pigment A, paeoniflorin, forsythiaside A, ferulic acid, berberine and glycyrrhizin acid in one run by HPLC.Their average contents and RSD in each Pifubingxuedu tablet were 0.299 5 mg/tablet (2.25％);0.700 0 mg/tablet(1.33％);0.429 2 mg/tablet (1.21％);0.039 1 mg/tablet (2.34％);0.014 8 mg/tablet(2.23％);0.209 0 mg/tablet (2.06％);0.272 7 mg/tablet (2.68％), respectively.Conclusion The established method is simple, convenient, accurate and reliable.It is suitable for the quality control of Pifubingxuedu tablet.

Effect of deactivation of enzymes on chemical composition of Forsythiae Fructus using 1H-NMR-based metabolomics approach / 中草药

Objective: To investigate the effect of deactivation of enzymes on chemical composition of Forsythiae Fructus using NMR based metabolomics approach. Methods: 1H-NMR-based metabolomics approach combined with multivariate statistical analysis was used to investigate the differential metabolites between raw Forsythiae Fructus (RF) and Forsythiae Fructus with deactivation of enzymes (DF). And degradation pathway of significant metabolites was inferred by biosynthetic pathway. Results: Twenty-four metabolites were identified in the 1H-NMR spectrum of Forsythiae Fructus, and principal component analysis (PCA) showed clear separation between RF and DF. The S-plot of orthogonal partial least square discriminate analysis (OPLS-DA) revealed that ten compounds contributed to the separation of RF and DF, and the levels of forsythiaside A, forsythiaside C, phillyrin, rutin, and surcose were higher in DF than those in RF, while the levels of rengyol, rengyoside, rengyolone, α-glucose, and β-glucose were higher in RF than those in DF. Moreover, the degradation of forsythiaside A into rengyol was proposed. Conclusion: This study reveals the chemical effect of deactivation of enzymes on Forsythiae Fructus in a holistic manner, which confirms rationality and necessity of deactivation of enzymes during the processing of Forsythiae Fructus. This study also serves as a basis for the bioactive compounds and quality standards research of Forsythiae Fructus.

HPLC fingerprint of Shufeng Jiedu Capsule / 中草药

Objective: To establish an HPLC method for the fingerprint analysis of Shufeng Jiedu Capsule (SJC), so as to provide evidence for the quality control of it. Methods: HPLC method was applied with the chromatographic condition as follows: The chromatographic column was Unitary C18 column (200 mm×4.6 mm, 5 μm), acetonitrile-0.1% formic acid as the mobile phase with gradient elution, the flow rate was 1.0 mL/min, the detection wavelength was 250 nm, the column temperature was 30℃, and the injection volume was 10 μL. The HPLC fingerprint of SJC was analyzed with similarity analysis and principal component analysis (PCA) method. Results: The fingerprint chromatography included 22 mutual peaks, of which eight mutual peaks from Polygonum cuspidatum, six mutual peaks from Forsythia suspensa, three mutual peaks from Verbena officinalis, two mutual peaks from Glycyrrhiza uralensis, and one mutual peak from Thlaspi arvense. The similarity among 14 batches was more than 0.9.Based on the HPLC-MS2 data, 16 components were identified, which were forsythosid E, hastatoside, verbenalin, 5'-hydroxy-forsythiaside A, polydatin, phillyrin, forsythosid A, acteoside, iso-forsythosid A, aloe-emodin, emodin-8-O-grapeglycosides, parietic acid, monoammonium glycyrrhizinate, 3-hydroxyglabrol, emodin, and chrysophanol. Conclusion: It is the first time to establish the HPLC fingerprint of SJC. The method is simple, accurate, and reproducible, which could reflect the chemical composition information of SJC comprehensively and provide scientific evidence for the quality control.

Study on Extraction Process of Xiaoer Yinqiao Granules / 中国药师

Objective:To establish the optimal extraction technology of Xiaoer Yinqiao granules by orthogonal test. Methods:The effects of water amount,extraction duration and extraction times were investigated by orthogonal design using the contents of forsythia-side A and chlorogenic acid as the indices. Results: The optimum extraction process was as follows: adding 8-fold amount of water, and extracting 1. 5 h for the first time, and then adding 6-fold amount of water, extracting 1 h for the second and third time, respective-ly. Conclusion:The extraction technology is simple, reasonable and reliable.

Simultaneous Determination of the Contents of 8 Ingredients of Shuanghuanglian Freeze-dried Powder in Pla-cental Perfusate / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of the contents of Shuanghuanglian freeze-dried powder in placental perfusate. METHODS:HPLC was performed on the column of Agilent Zorbax-C18 with mobile phase of acetonitrile-1%formic acid aqueous solution(gradient elution)at flow rate of 1.0 ml/min,the internal standard was puera-rin,detection wavelength was 280 nm,column temperature was 25 ℃ and sample volume was 10 μl. RESULTS:There was a lin-ear range between linear ranges and peak area of 8 ingredients(r≥0.999 0);RSDs of within-day and inter-day precision tests were no more than 1.9%,repeatability tests was no more than 7.3%;average recoveries were in the range of 92.73%-112.37%(RSD=3.2%-8.2%,n=6);and average matrix effects were 90.33%-105.78%(RSD=3.2%-8.0%,n=6). CONCLUSIONS:The method is rapid,sensitive and specific,and can be used to the simultaneous determination of the contents of 8 ingredients of Shuan-ghuanglian freeze-dried powder in placental perfusate.

Optimization of ultrahigh pressure extraction process of chlorogenic acid and forsythiaside in Jinqiao Reduqing granules by Box-Behnken design-response surface methodology / 中国药学杂志

OBJECTIVE: To optimize the ultrahigh pressure extraction (UPE) process of effective constituents from Jinqiao Reduqing granules by Box-Behnken design-response surface methodology. METHODS: On the basis of single factor screening, a three-factor and three-level Box-Behnken experimental design was employed, with solvent ratio (X1), extraction pressure (X2) and extraction time(X3) as independent variables. Dependent variables were transformed into desirabilities mathematically by Hassan's method. Data of overall desirabilities were fitted to a second order polynomial equation, through which three dimensional response surface graphs were produced. Optimum experimental conditions were selected from the stationary point of the response surfaces. RESULTS: Regression coefficient of binomial fitting complex model was as high as 0.9878. The optimum conditions of UPE were as follows: the materials were extracted for 38 s with 24 times amount of water, the extraction pressure was 112.1 MPa, and the extraction was performed for one tine. The bias between observed and predicted values was 5.27%. CONCLUSION: As a novel extraction technology for Chinese herbal medicine, the UPE procedure has higher extraction yield, lower extracting temperature, shorter extracting time and less power consumption. The UPE has provided a brand new method for extraction of chlorogenic acid and forsythiaside from Jinqiao Reduqing granules.

Separation and purification of phenylethanoid glycosides from Callicarpa kwangtungensis by macroporous resin / 中草药

Objective: To study the technology for separation and purification of phenylethanoid glycosides from Callicarpa kwangtungensis by macroporous resin. Methods: The absorption and separation properties of 10 kinds of absorption resins were studied and the separation and purification technological process of phenylethanoid glycosides from C. kwangtungensis were optimized by HPLC with the total content of forsythiaside B and poliumoside as an index. Results: Using X-5 resin column as adsorbent, the optional conditions were as follows: loading qualities were 32.79 mg/g, the resin was washed by 4 BV distilled water to remove impurity and 7 BV 30% ethanol to elute phenylethanoid glycosides, the eluting velocity was 2 BV/h, and the content of phenylethanoid glycosides in the extract was 52.8% (n = 3). Conclusion: This technology is simple and feasible, and the process with X-5 resin is an effective method to separate and purify phenylethanoid glycosides from C. kwangtungensis.

Determination of nine components in Forsythia Folium collected in different periods / 中草药

Objective: To develop an HPLC-MS method for the determination of phillyrin, phillygenin, forsythiaside A, forsythiaside F, rutin, quercetin, isoquercitrin, chlorogenic acid, and (+)-epipinoresinol-4'-O-glucoside in Forsythia Folium. Methods: The samples were separated on a Dikma Diamonsil C18 column (150 mm × 4.6 mm, 5 μm), eluted with methanol and water containing 0.02% formic acid in a gradient elution at a flow rate of 0.8 mL/min, and the injection volume was 10 μL. The column temperature was set at 30°C. The multiple-reaction monitoring scanning was employed for the quantification with switching electrospray ion source polarity in negative mode. The ion spray voltage was set at -4500 V and the turbo spray temperature was maintained at 650°C. Results: The regression equations showing linear relationships between peak areas and contents of each compound were obtained. The average recoveries of the compounds ranged from 97.20% to 101.2% and the precision in terms of RSD was in the range of 0.54%-1.24%. Conclusion: The method is sensitive and suitable for the determination of the nine components in Forsythia Folium. As a result, the suitable harvesting time for Forsythia Folium is May.

Comparison of Active Components of Different Positions and Different Processing Methods / 中国中医药信息杂志

Objective To study the components of Forsythia suspensa in stalks and leaves, and compare the content between the crude-sun-cured and the steam-sun-cured. Methods The components and content in stalks and leaves to fruits of Forsythia suspensa were compared by RP-HPLC. Results There were some same active components in stalks, leaves and fruits, and the steam-sun-cured had higher content. Conclusion The steam-sun-cured stalks and leaves of Forsythia suspensa could be used to extract components and develop the tea.

Determination of Forsythiaside and Forsythin from Fruits of Forsythia Suspensa and Forsythia Viridissima Lindl Collected in Different Periods / 中国中医药信息杂志

Objective To study the content of forsythiaside and forsythin from fruits of Forsythia suspensa and Forsythia viridissima Lindl collected in different periods. Methods Samples were dealt by HPLC-PDA with Diamonsil-C18 (4.6 mm?250 mm,5 ?m) column. The mobile phase consisted of methanol-water,gradient elution,the flow rate was 1.0 mL/min. The UV detection was set at 270 nm. Results The content of forsythiaside from fruits of Forsythia suspensa was much higher than that from fruits of Forsythia viridissima Lindl. And there was no forsythin from fruits of Forsythia viridissima Lindl. Conclusion There was much difference in content of forsythiaside and forsythin from the two kinds of fruits above. Forsythia viridissima Lindl should not be used as Forsythia suspensa.

Regulation of dry substance and active components accumulation in fruit of Forsythia suspensa / 中草药

Objective To study the regulation of dry substance and active components accumulation in the fruit of Forsythia suspensa. Methods The dry substance of the fruit was analyzed by the method of hundred grain mass. The contents of the active components in leaves, shucks, and seeds were analyzed by HPLC. Results The period for F. suspensa to accumulate the dry substance was mainly May and July. During the whole growth phase, the active components, such as forsythiaside, forsythin, and rutin in different parts, were decreasing. From the last ten-day of August to the first ten-day of September, the accumulation of the dry substance came to stability and the contents of active components in shucks and seed were stable. The contents of active components in shucks decreased rapidly from September to October, and then came to stability and the contents in shucks is the least during the last ten-day of October. So the contents of active components in younger F. suspensa are higher than that in elder one, and the proportion among forsythiaside, forsythins, and lution is also significantly different from elder seeds. Conclusion It is more preferable to pick younger F. suspensa from the last ten-day of August to the first ten-day of September and to pick elder F. suspensa during last ten-day of October.