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Objective: To investigate the mechanism of forsythin in inhibiting the growth, migration and invasion of human renal cancer cells (786-0). Methods Human renal cancer cells (786-0) were cultured in vitro, and different concentrations of forsythin were added. Cell viability was detected by MTT assay, cell apoptosis was detected by AO/EB assay, cell migration and invasion abilities were respectively investigated by wound healing and transwell migration assays. The expression levels of PI3K, p-PI3K, Akt, p-Akt, FOXO3a, p-FOXO3a, p21, p27, Fasl, Bim, MMP-2, and MMP-9 were detected by Western bloting. Results:In 786-0 cells, forsythin effectively inhibited the growth of renal cancer cells, promoted apoptosis, interfered with cell cycle, and upregulated expression levels of p21, p27, Fasl, and Bim, when compared with control group (P < 0.05, 0.01); Compared with the control group, different concentrations of forsythin can significantly inhibit the migration and invasion of renal cancer cells, and reduce the synthesis of MMP-2 and MMP-9 (P < 0.05, 0.01). Meanwhile, forsythin can significantly inhibit the phosphorylation of PI3K, Akt and FOXO3a in a concentration-dependent manner (P < 0.05, 0.01). Conclusion: Forsythin can regulate apoptosis and cell cycle, and inhibit the growth of renal cancer cells by down-regulating the PI3K/Akt signaling pathway. At the same time, forsythin can effectively inhibit the ability of renal cell migration and invasion through the PI3K/Akt pathway.
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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.
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To establish a method for the simultaneous determination of 9 components, namely gardenoside, paeoniflorin, forsythoside A, baicalin, forsythin, glycyrrhizic acid, rhein, honokiol, and magnolol in Xiao'er Chiqiao Qingre granules（XECQ Gra）. Ultra performance liquid chromatography (UPLC) was used on an Acquity UPLC HSS T3 C₁₈ column (2.1 mm×100 mm, 1.8 μm) with 0.1% phosphoric acid acetonitrile (A)-0.1% phosphoric acid solution (B) as mobile phase for gradient elution. The flow rate was 0.3 mL·min⁻¹ ; the column temperature was set at 30 °C, and the determination wavelength was set at 220 nm. All the 9 compounds were well separated, and showed good linear relationship within their concentrations (r>=0.999). The average recoveries were between 95.84%-101.4% and the RSD values were all less than 3.0%. The method is simple, reliable, and accurate, and could be used for the quality control of XECQ Gra.
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In order to obtain the optimum method for content determination of Forsythia Fructus (FF), a variety methods for the sample preparation of FF were evaluated by the content determination methods of Chinese Pharmacopoeia. And an optimum method was screened and as follows: 30 times with 70% ethanol solution in ultrasonic extractor for half an hour. The method can achieve the best effect of simultaneously extracting forsythoside A and forsythin. Then, a HPLC method for simultaneous determination of forsythoside A and forsythin was established by methodology. The HPLC chromatographic conditions: the mobile phase consisted of acetonitrile (A)-0.4% acetic acid solution (B) with gradient elution [0-33 min，15%A，33-43 min，15%-25%A，43-60 min，25% A] was at the flow rate of 1 mL·min⁻¹, the column temperature was 25 °C, and the detection wavelength was 330 and 277 nm. Moreover, the contents of forsythoside A and forsythin for 10 Green Forsythia Fructus (GF) and 5 Old Forsythia Fructus (OF) were determined by this method and Chinese Pharmacopoeia. The result not only displayed that the established method is effective, rapid, and simple, but also showed that the contents of forsythoside A and forsythin for GF and OF were significantly different. Which implied that the forsythoside A and forsythin limit standard for GF and OF should be controled by different values. This studies provide an important basis for the establishment of the content determination of FF and the quality control standard for GF and OF.
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OBJECTIVE@#To access the toxicity of forsythin from Forsythia suspensa leaves and evaluate its safety.@*METHODS@#Acute toxicity was determined by oral administration of a single dose of 18100 mg/kg forsythin in NIH mice. Sub-chronic toxicity was evaluated by oral administration of several doses of forsythin for 30 days at does of 0, 540, 1620, and 6480 mg/kg in SD rats.@*RESULTS@#In the acute toxicity study, mortality was not observed after 14 days. In addition, clinically relevant adverse effects, or variations in body weight or food consumption were not observed. Similarly, after 30 days in the sub-chronic toxicity study, no mortality or significant toxicological effects such as decreased food consumption, body weight, biochemical parameters and vital organs etc. were noticed.@*CONCLUSION@#The results revealed that the forsythin from Forsythia suspensa leaves has low or no toxicity via oral administration, and therefore is suitable for further development and applications.
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OBJECTIVE:To establish the method for simultaneous determinations of rutin,forsythin and platycodin D in Sangju ganmao pills.METHODS:HPLC-MS method was adopted.The determination was performed on Waters Atlantis C18 column with mobile phase consisted of acetonitrile-0.1％ formic acid (gradient elution) at the flow rate of 0.2 mL/min.The column temperature was set 35 ℃,and sample size was 10 μL.The ionization mode was electrospray ion,and the reaction mode was multi-reaction monitoring.By positive ion detection mode,the drying gas and nebuliser gas were all high purity nitrogen.The drying gas temperature was 270 ℃.The drying gas flow rate was 25 L/min.The sheath gas flow rate was 10 L/min.The capillary voltage was 4 500 V.The nozzle voltage was 2 000 V and the scanning time was 0.1 s.RESULTS:The linear range of rutin,forsythin and platycodin D were 0.010 82-2.164 μg/mL (r=0.999 7),0.010 18-2.036 μg/mL (r=0.999 4),0.010 27-2.054 μg/mL (r=0.999 7),respectively The limits of quantification were 1.250,0.260,2.720 ng/mL,and the limits of detection were 0.380,0.078,0.820 ng/mL.RSDs of precision,stability and reproducibility tests were all no more than 3.0％.The recoveries were 97.88％-99.88％ (RSD=0.72％,n=6),98.48％-103.13％ (RSD=1.91％,n=6),98.79％-101.41％ (RSD=1.05％,n=6).CONCLUSIONS:This method is simple,precise,stable and reproducible,and can be used for simultaneous determination of rutin,forsythin and platycodin D in Sangju ganmao pills.
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Objective To access the toxicity of forsythin from Forsythia suspensa leaves and evaluate its safety. Methods Acute toxicity was determined by oral administration of a single dose of 18 100 mg/kg forsythin in NIH mice. Sub-chronic toxicity was evaluated by oral administration of several doses of forsythin for 30 days at does of 0, 540, 1 620, and 6 480 mg/kg in SD rats. Results In the acute toxicity study, mortality was not observed after 14 days. In addition, clinically relevant adverse effects, or variations in body weight or food consumption were not observed. Similarly, after 30 days in the sub-chronic toxicity study, no mortality or significant toxicological effects such as decreased food consumption, body weight, biochemical parameters and vital organs etc. were noticed. Conclusion The results revealed that the forsythin from Forsythia suspensa leaves has low or no toxicity via oral administration, and therefore is suitable for further development and applications.
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Objective To establish the fingerprint of Forsythia suspensa dispensing granule by HPLC. Methods The total of 10 samples of Forsythia suspensa were analyzed by HPLC on a Waters SunFire C18 column(4. 6 mm×250 mm,5 μm) with mobile phase of A:acetonitrile and mobile phase B:0. 4% acetic acid solution as gradient elution, with flow rate at 1. 0 mL·min-1 , wavelength at 277 nm and column temperature at 30 ℃. Results There were 13 characteristic common peaks in the Forsythia suspensa dispensing granule, in which two peaks identified as forsythoside A and forsythin, the similarities were more than 0. 999. Conclusion The HPLC fingerprint of Forsythia suspensa dispensing granule is stable, and is easy to manipulate, which can provide more information for the quality control of Forsythia suspensa dispensing granule.
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Objective: To develop an HPLC method for the simultaneous determination of five active components ( 5-O-methyl-visammioside, prim-O-glucosycimifugin, hesperidin, rosmarinic acid and forsythin) in Sijiganmao tablets. Methods:The samples were separated on an Agilent C18 column (250 mm × 4. 6 mm,5 μm) by acetonitrile-0. 1% formic acid solution with gradient elution. The detection wavelength was 283nm,the flow rate was 1. 0μl·min-1 ,the column temperature was 30℃,the detection wavelength was 283 nm,and the sample size was 10μl. Results:The complete separation was obtained for the five active compounds. The five regression e-quations showed good linear relationships. The average recoveries of the compounds were between 95. 0% and 105. 0%. Conclusion:The established method is accurate, reliable, simple and effective, which can be used in the quality control of Sijiganmao tablets.
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Objective:To establish the HPLC fingerprints of chemical constituents in Forsytiae suspensa Fructus from Shiyan dis-trict to provide scientific evidence for the quality control of Forsytiae suspensa Fructus. Methods: HPLC was performed on a Waters SunFire C18 (250 mm × 4. 6 mm, 5 μm) column and a Diamonsil C18 guard column, and the mobile phase consisted of acetonitrile-0. 4% acetic acid solution with gradient elution. The flow rate was 1 ml·min-1 , the detection wavelength was 277 nm, and the col-umn temperature was 30℃. Results:There were 14 common peaks in the HPLC fingerprints of chemical constituents in 12 batches of Forsytiae suspensa Fructus from Shiyan. The 14 characteristic peaks of Forsytiae suspensa Fructus from different habitats were under clustering analysis, and the similarity showed little difference. The contents of forsythoside A and forsythin exhibited significant differ-ence in the samples. Conclusion:The HPLC fingerprint method is simple, rapid and feasible in the quality control of Forsytiae suspen-sa Fructus.
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Objective To establish a new RP-HPLC method for simultaneous determination of chlorogenic acid,forsythin, and baicalin in shuanghuanglian powder for injection after ultrasonic atomization. Methods Hypersil ODS2 C18(250 mmí4. 6 mm,5 μm) was used as the chromatographic column. The mobile phase was methanol-0. 2% phosphate acid solution (4060). Flow rate was 1. 0 mL·min-1 . Sample volume was 5μL. Column temperature was 30℃. Detection wavelength was 324 nm at 0-10 min and 277 nm at 10-25 min. Results Contents of chlorogenic acid,forsythin, and baicalin had good linear relationship with the respective peak area (r≥0. 999 7) within the scope of the sample volume. The RSD was <2% for precision, reproducibility, and stability. Recovery rate was 98. 50%-101. 12% (n=6). Conclusion The method is rapid, accurate and reproducible, with high resolution. It can determine the content of three kinds of components at the same time. The three components in shuanghuanglian powder for injection did not change significantly before and after ultrasonic atomization.
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Objective To compare the contents of forsythoside A and forsythin in Fructus Forsythia and its dispensing granule. Methods HPLC-gradient elution method was used with SunFire C18 column (4.6 mm×250 mm, 5μm), mobile phase A of acetonitrile and B of acetic acid, flow rate of 1.0 mL/min, detection wavelength of 277 nm, and column temperature at 30 ℃. HPLC was used to determine the contents of forsythoside A and forsythin in Fructus Forsythia and its dispensing granule, and compare the difference between the two contents. Results The content of forsythoside A in dispensing granule was less than that of raw material of Fructus Forsythia, and the concentration of the major components in the commercial Lianqiao Granule were not equivalent to that in the decoction of Fructus Forsythia. The content of forsythin in dispensing granule was equivalent with that of raw material of Fructus Forsythia. Conclusion The original formula granule production process needs to be improved, and the standardized criteria for the quality control and reasonable quality standard of granule should be established.
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Objective: To establish an HPLC method for the simultaneous determination of cafferic acid, forsythoside A, forsythoside B, rutin, hyperoside, forsythin, and arctigenin in Forsythia suspensa. Methods: The analysis was carried out on an Inertsil ODS-3 C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase was composed of acetonitrile and 0.2% phosphorie acid aqueous with gradient elution. The detection wavelength was set at 275 nm. The flow rate was 1.0 mL/min at column temperature of 30 °C. Results: Cafferic acid, forsythoside A, forsythoside B, rutin, hyperoside, forsythin, and arctigenin were well separated by this method, and showed a good linearity in the ranges of 18.24-91.20, 5.88-29.40, 132.60-663.00, 8.34-41.70, 1.96-9.80, 7.60-38.00, and 11.34-56.70 μg/mL, respectively. The average recoveries of the seven components were 97.7%, 96.7%, 102.6%, 101.3%, 93.2%, 91.8%, and 96.7% and the RSD values were 2.3%, 1.4%, 2.4%, 2.2%, 1.0%, 1.0%, and 1.3%, respectively. Conclusion: The established method is accurate, reliable, and could be used for the simultaneous determination of the seven components in F. suspense, which provides a scientific basis for the quality evaluation of F. suspense.
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Objective: To compare the two parameters in Raman spectroscopy quantitative analysis, so as to confirm the optimal one. Methods: The ratios of peak intensity and peak area were taken as parameters in quantitative analysis. The contents of forsythin in forsythia suspense leaves gathered at different time points from different regions were analyzed with laser Raman spectroscopy. When the ratio of the peak intensity was taken as parameter, the Raman peak of forsythin (ρΛr-H [1 319.1 cm-1]) and the peak of methanol (νasCH3-O [2 974. 7 cm-1]) were selected as quantitative peak and internal standard reference peak in the confocal micro-Raman spectra of forsythin methanol solution. When the ratio of the peak area was taken as parameter, the Raman peak of forsythin ( ρΛr-H [1 319.1 cm-1]) and the peak of methanol (νsCH3-O[3 020. 9 cm-1]) were selected. The ratios of intensity and area were taken as ordinate, and the content of forsythin was taken as abscissa. The two standard curves were plotted and the linear and recoveries were compared. Results: Both parameters had good linear relationship with forsythin concentration, with the correlation coefficients being 0.998 0 and 0.997 6; the recoveries were 100.04%-101.30% and the recycling was complete. The linear fitting of the results with two parameters were measured, and the regression equation was C2=1.030 4C 1=0.033 1, r=0.999 6. The results obtained with the two parameters were accurate and were largely identical. Conclusion: Both the relative peak intensity and the ratio of the peak area can be used as parameters for forsythin quantitative analysis with Raman spectroscopy. And the two parameters can obtain largely identical results. Raman spectroscopy with internal standard is a simple and rapid method for quantitative analysis of forsythin, and it can be used for quantitative analysis for Chinese herbs.
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OBJECTIVE: To study the quality of Shuanghuanglian oral solutions from different manufactures. METHODS: The contents of baicalin, chlorogenic acid and forsythin in Shuanghuanglian oral solution on the market were determined by HPLC. The determination was performed on Agilent ZORBAX 80A Extend-C18 (150 mm?4.6 mm,5 ?m) column with column temperature set at 30 ℃. The mobile phase for baicalin consisted of methanol-water-acetic acid (50 ∶ 50 ∶ 1, V/V) at flow rate of 1.0 mL?min-1 and detection wavelength of 274 nm. The mobile phase for chlorogenic acid consisted of methanol-water-acetic acid (20 ∶ 80 ∶ 1, V/V) at flow rate of 0.8 mL?min-1 and detection wavelength of 324 nm. The mobile phase for forsythin consisted of acetonifrile-water (25 ∶ 75, V/V) at flow rate of 1.0 mL?min-1 and detection wavelength of 278 nm. RESULTS: Shuanghuanglian oral solutions from different manufactures conformed to the requirements of the first column of Chinese Pharmacopeia (2005 edition). The content differences of chlorogenic acid in Shuanghuanglian oral solutions were almost 3 times, which may be the cause of the difference of clinical efficacy. CONCLUSION: The minimum detection limit only has been established for the quality standard of Shuanghuanglian oral solution. It is suggested to improve the quality standard of Shuanghuanglian oral solution.
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Objective To establish a method for the determination of Forsythin in Shenyan Jiere Tablets by HPLC. Methods The separation was performed on shim-pak clc-ODS (4.6 mm?150 mm, 5 ?m) with Accetonitrile-0.025 mol/L H3PO4 (20∶80) as mobile phase. The flow rate was 1 mL/min and UV detector was at 277 nm. Results The method had good average recovery with 97.1% (n=5), RSD=1.6%, good linear relationship within the range of 0.12～0.5 ?g. Conclusion The method is accurate, reliable and can be used for quality control of the production.
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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.
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Objective To establish a HPLC method for the determination of chlorogenic acid, baicaliin, forsythin and wogonin in Shuanghuanglian oral liquid. Methods The column was VP-ODS C18 (250 mm?4.6 mm, 5 ?m). The mobile phase consisted of acetonitrile-0.2% phosphoric acid with gradient elution. The flow rate was 1.0 mL/min and the detection wavelength was at 278 nm. Results The calibration curves were linear within the range of 0.44～6.60 ?g (r=0.999 1) for chlorogenic acid, 0.52～6.18 ?g (r=0.999 1) for baicaliin, 0.20～2.04 ?g (r =0.999 3) for forsythin (r =0.999 3) and 0.13～1.76 ?g (r =0.999 1) for wogonin, respectively. The average recovery of them were 97.0% (RSD=1.1%), 98.98% (RSD=1.1%), 103.55% (RSD = 1.1%) and 96.49% (RSD = 1.1%), respectively. Conclusion The method is simple, practicable, accurate and rapid. It can be applied to content determination of chlorogenic acid, baicaliin, forsythin and wogonin in Shuanghuanglian oral liquid.
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OBJECTIVE:To estabish a RP-HPLC method for the content determination of forsythin of Yinqiao jiedu pills.METHODS:The determination was performed using SHIM-PACK VP-ODS column(150 mm ? 4.6 mm,5 ?m) with column temperature at 30 ℃.The mobile phase was acetonitril-water(25∶75) with flow rate at 1.0 mL?min-1.The detective wavelength was set at 277 nm.RESULTS:The linear range of forsythin was 0.453～3.481 ?g(r=0.999 8).The mean recovery was 99.50%(RSD=1.15%,n=6).CONCLUSION:The method was simple,rapid,accurate and suitable for the quality control of Yinqiao jiedu pills.
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OBJECTIVE:To establish a HPLC method for content determination of forsythin in Qingrejiedu granules.ME_ THODS:A Alltech Inersil ODS-2column(150mm?4.6mm,5?m)was used,the mobile phase was acetonitrile-water(18∶32), the detective wavelength was277nm,the column temperature was30℃,the flow rate was1.0ml/min.RESULTS:The linear range of forsythin was0.0984?g～12.3?g(r=0.9997),the average recovery was100.62%(RSD=0.6%).CONCLUSION:This method was easy to operate,good reproducibility and accurate result,it can be used for quality control in production Qin?grejiedu granules.