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The NLRP3 inflammasome's core and most specific protein, NLRP3, has a variety of functions in inflammation-driven diseases. Costunolide (COS) is the major active ingredient of the traditional Chinese medicinal herb Saussurea lappa and has anti-inflammatory activity, but the principal mechanism and molecular target of COS remain unclear. Here, we show that COS covalently binds to cysteine 598 in NACHT domain of NLRP3, altering the ATPase activity and assembly of NLRP3 inflammasome. We declare COS's great anti-inflammasome efficacy in macrophages and disease models of gouty arthritis and ulcerative colitis via inhibiting NLRP3 inflammasome activation. We also reveal that the α-methylene-γ-butyrolactone motif in sesquiterpene lactone is the certain active group in inhibiting NLRP3 activation. Taken together, NLRP3 is identified as a direct target of COS for its anti-inflammasome activity. COS, especially the α-methylene-γ-butyrolactone motif in COS structure, might be used to design and produce novel NLRP3 inhibitors as a lead compound.
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OBJECTIVE：To study the effects of costunolide （COS）on the cell cycle distribution and apoptosis of human breast cancer SK-BR- 3 cells and its mechanism. METHODS ：Human breast cancer SK-BR- 3 cells were divided into blank control group，and COS groups of 10，20，30，40，50 μmol/L. MTT assay was used to detect the effects of COS on cell proliferation. SK-BR-3 cells were divided into blank control group ，COS low ，medium and high concentration groups （10，20，30 μmol/L）. After cultured for 24 h，flow cytometry was used to detect the distribution of cell cycle. Hoechst 33258 fluorescence staining was used to detect cell apoptosis. Western blot assay was used to detect the expression of p 53，caspase-3，Bcl-2，Bax，p21，CDK2 and cyclinE. RESULTS ：Compared with blank control group ，COS could significantly inhibit the proliferation of SK-BR- 3 cells（P＜ 0.05 or P＜0.01），and in a dose and time-dependent manner. Low ，medium and high concentrations of COS could induce cell apoptosis and arrest cell at G 1/S phase （P＜0.05 or P＜0.01），could significantly up-regulate the protein expression of p 53， caspase-3，Bax and p 21（P＜0.05 or P＜0.01），and could significantly down-regulate the protein expression of Bcl- 2，CDK2 and cyclinE（P＜0.01）. CONCLUSIONS ：COS can inhibit the proliferation of human breast cancer SK-BR- 3 cells and induce cell apoptosis and cell cycle arrest. The mechanism may be related to the regulation of p 53/Bax/Bcl-2/caspase-3 apoptosis signal pathway.
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Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) method for the simultaneous determination of six components of gallic acid, hydroxysafflor yellow A, geniposide, ellagic acid, costunolide and dehydrocostus lactone in Gurigumu-13 Pill, which is proved to be a scientific and feasible method in the quality analysis in Gurigumu-13 Pill. Methods: The relative factor (fs/i) of gallic acid, ellagic acid, hydroxysafflor yellow A, costunolide and dehydrocostus lactone were established by HPLC method with geniposide as internal standard, which were used to calculate the content of five constituents in the samples of Gurigumu-13 Pill. Meanwhile, external standard method (ESM) was used to calculate the content of six constituents. The difference between QAMS and ESM was analyzed to evaluate the accuracy of QAMS. Results: The fs/i of gallic acid, hydroxysafflor yellow A, ellagic acid, costunolide and dehydrocostus lactone were 0.481 0, 0.906 4, 0.170 9, 0.971 2 and 1.261 5, respectively. The content determination results of six batches of Gurigumu-13 Pill were calculated by the method of QAMS and ESM, with no significant difference in RSD < 2.0%. Conclusion: The fs/i established in the QAMS method with geniposide as the internal reference substance is accurate and simple. The QAMS method can be used for the multi-index quality evaluation of Gurigumu-13 Pill.
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Objective: To develop and validate an high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of 13 major bioactive components (ferulic acid, costunolide, baicalin, paeoniflorin, tetrahydropalmatine, rutecarpine, berberine, palmatine, evodiamine, naringin, hesperidin, saikosaponin a, and saikosaponin d) in Jiawei Zuojin Pills (JZP). Methods: The chromatographic separation was performed on a Thermo Syncronis C18 column (100 mm × 4.6 mm, 3.0 μm) with a gradient elution of methanol and 0.02 mol/L ammonium acetate in water at a flow rate of 0.35 mL/min, and the injection volume was 10 μL. The 13 major bioactive components were detected using an electrospray ionization source in ESI+ and ESI- ionization mode, quantified by multiple reaction monitor (MRM) scanning at the same time. Results: The linear ranges of ferulic acid, costunolide, baicalin, paeoniflorin, tetrahydropalmatine, rutecarpine, berberine, palmatine, evodiamine, naringin, hesperidin, saikosaponin a and saikosaponin d were 2-80 μg/mL (r = 0.999 2), 0.5-20.0 μg/mL (r = 0.999 1), 3.5-140.0 μg/mL (r = 0.999 8), 1-40 μg/mL (r = 0.999 2), 0.3-12.0 μg/mL (r = 0.999 1), 1-40 μg/mL (r = 0.999 2), 3-120 μg/mL (r = 0.999 7), 2.5-100.0 μg/mL (r = 0.999 5), 0.5-20.0 μg/mL (r = 0.999 3), 0.5-20.0 μg/mL (r = 0.999 1), 1-40 μg/mL (r = 0.999 1), 0.3-12.0 μg/mL (r = 0.999 2), 0.3-12.0 μg/mL (r = 0.999 2), and the average recoveries were 99.5% (RSD = 4.11%), 98.9% (RSD = 4.88%), 100.2% (RSD = 1.08%), 99.2% (RSD = 3.23%), 99.5% (RSD = 4.13%), 99.7% (RSD = 3.23%), 98.6% (RSD = 2.78%), 99.9% (RSD = 3.12%), 101.3% (RSD = 4.53%), 98.7% (RSD = 3.43%), 99.8% (RSD = 3.58%), 97.9% (RSD = 5.22%), and 101.3% (RSD = 5.13%), respectively. The contents of 12 batches of the 13 major bioactive components were 0.324-0.383, 0.051-0.072, 3.225-3.466, 0.154-0.198, 0.015-0.062, 0.144-0.199, 2.145-2.982, 0.441-0.953, 0.032-0.099, 0.062-0.089, 0.111-0.178, 0.012-0.065, 0.011-0.069 mg/g, respectively. Conclusion: The developed method is simple, specific, and sensitive, and it can be applied for the determination of 13 major bioactive components and the quality control of JZP.
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Objective:To explore the synergistic effect of arbuscular mycorrhizal（AM） fungi mixed inoculation on the growth，physiological and biochemical characteristics，root biomass and terpenoid component accumulation of Aucklandia lappa seedlings，so as to provide a reference for the combination and application of the dominant complementary effect mycorrhizal fungi. Method:The effect of different AM fungi combined with inoculation on the root mycorrhizal infection rate，plant growth，physiological and biochemical characteristics，root biomass，costunolide and dehydrocostus lactone of A. lappa seedlings were determined by pot inoculation at room temperature. Result:It was found that AM fungi could form good mycorrhizal symbiosis with the roots of A. lappa.The formation of mycorrhizal symbiosis system could increase the chlorophyll content of A. lappa leaves，increase the activities of catalase（CAT），peroxidase （POD），superoxide dismutase （SOD），reduce the content of malondialdehyde（MDA），and promote photosynthesis of A. lappa. Compared with CK group，AM fungus treatment could significantly promote the accumulation of costunolide and dehydrocostus lactone，and the accumulation of its metabolites，costunolide and dehydrocostus lactone，into roots during the symbiotic cultivation of A. lappa seedlings，indirectly improving the quality of medicines and yield of alantolactone. Conclusion:Inoculation of AM fungi can improve the root mycorrhizal viability，increase the absorption of nutrients and promote the growth of woody incense.The mixed inoculation treatment of S2，S4 and S5 had the best mycorrhizal effect in artificial cultivation，and the growth and medicinal quality of A. lappa were the best，which provided technical support for the application and popularization of A. lappa mycorrhizal biotechnology.
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OBJECTIVE：To study the effects of costunolide on the proliferation ，migration and apoptosis of breast cancer SK-BR-3 cells and its mechanism. METHODS ：SK-BR-3 cells in logarithmic growth period were collected and cultured with different concentrations （10，20，30，40，50 μmol/L）of costunolide for 24，48，72 h. Inhibitory rate of costunolide on cell proliferation was detected with CCK- 8. The cells were divided into blank control group and costunolide group （10，20，30 μmol/L）. Hoechst 33258 fluorescence was used to observe the morphology and apoptosis of cells ，and apoptotic rate of cells were calculated. Cell scratch test was used to detect the migration ability of cells and calculate the migration rate. Western blotting was used to detect the relative expression level of Bcl- 2，Bax，Caspase-3 and Cleaved Caspase- 3 in cells. RESULTS ：The proliferation of SK-BR-3 cells were significantly inhibited by costunolide （P＜0.05 or P＜0.01），and it shows a trend of concentration and time dependence. In the blank control group ，cells possessed clear contour ，regular shape and good adherence . Compared with blank control group，the number of cells were decreased significantly in 10，20，30 μmol/L costunolide groups，the cell structure was loose，the volume was reduced ，and the gap became larger ，and most of the cell contour disappeared and became round ，the cell adherence was poor ；cell migration rate and Bcl- 2 protein relative expression level were decreased significantly ，while apoptosis rate and the relative expression level of Bax ，Caspase-3 and Cleaved Caspase- 3 protein were significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS ： Costunolide can inhibit the proliferation and migration ，and induce apoptosis of human breast cancer SK-BR- 3 cells，mechanism of which may be through up-regulating the expression of Bax ，Caspase-3 and Cleaved Caspase- 3 while down-regulating the expression ofBcl-2.
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Objective:To establish a supercritical fluid chromatography(SFC) method for separating and purifying costunolide and dehydrocostus lactone in Aucklandiae Radix. Method:With supercritical carbon dioxide as the mobile phase,the effect of six factors, such as type of chromatographic columns,modifiers and modifiers ratio, flow rate of mobile phase,pressure and temperature, on the separation process of supercritical fluid chromatography were explored. The target components were separated and prepared by semi-preparative supercritical fluid chromatography. High performance liquid chromatography and nuclear magnetic resonance were used to analyze the components and study the thermodynamic regularity of the chromatographic process. Result:C18 column (10 mm×250 mm,5 μm) was adopted, with supercritical fluid dioxide as the mobile phase,the ratio of methanol was 0.13%,the flow rate was 12 mL·min-1,column pressure was 13 MPa,column temperature was 318℃, and detection wavelength was 225 nm. The sample was injected for 20 times,crude extract was 4 mg,and each target component was collected according to the chromatogram. Its purity was determined to be more than 99%by HPLC,and its structure was determined as costunolide and dehydrocostus lactone by NMR. Under this condition,the SFC separation process was normal-phase chromatography. Conclusion:The method can be used to prepare effective components of Aucklandiae Radix with a high purity and low solvent residue.
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Objective To develop an HPLC method for simultaneous determination of the eleven constituents (agaric-alcohol, naringin, hesperidin, neohesperidin, honokiol, emodin, magnolol, costunolide, dehydrocostus, chrysophanol, and physcion) in Chenxiang Huazhi Pills (CHP) by HPLC with gradient elution. Methods The chromatographic separation was performed on an Thermo Syncronis C18 column (4.6 mm × 250 mm, 5 μm) which was operated at 30 ℃. The mobile phase was a linear gradient prepared from water (A) and acetonitrile (B). The linear gradient elution program was programmed as follows: 0—10 min, 20% acetonitrile; 10—20 min, 20%—40% acetonitrile; 20—24 min, 40% acetonitrile; 24—26 min, 40%—52% acetonitrile; 26—30 min, 52% acetonitrile; 30—31 min, 52%—90% acetonitrile; 31—35 min, 90% acetonitrile; 35—40 min, 90%—100% acetonitrile; 40—43 min, 100% acetonitrile; 43—45 min, 100%—20% acetonitrile. The flow rate was 1 mL/min and the detection wavelength was 215 nm. Results The analysis permitted very good separation of eleven constituents within 43 min. A good linear relationship between the peak area and the injection volume was obtained. The ranges of the eleven constituents were 1.4—13.6, 10.0—200.0, 31.5—315.0,1.0—120.1, 1.8—50.6, 0.93—10.1, 1.8—30.0, 0.2—40.3, 1.8—18.1, 1.7—25.0, and 0.45—10.70 μg/mL. The average recoveries of eleven constituents in the samples were in the range of 98.90%—100.87%. The precision RSD of the peak areas of the 11 components ranged from 0.55%—1.54%; Eleven components had good stability within 30 h, and the concentration RSD of each component ranged from 0.75% to 1.94%; The repeatability RSD of each component ranged from 0.39% to 1.73%. The content of agaric-alcohol, naringin, hesperidin, neohesperidin, honokiol, emodin, magnolol, costunolide, dehydrocostus, chrysophanol, and physcion in six batches were 92.0—201.0, 511.5—9 033.0, 5 475.0—12 635.5, 54.5—5 095.5, 192.0—2 137.5, 117.0—391.5, 106.5—1 281.5, 13.0—136.5, 93.5—199.0, 177.0—1 207.0, and 33.5—251.5 μg/g, respectively. Conclusion The method is accurate, rapid and simple with high sensitivity, precision and repeatability, which has been successfully applied as an effective tool for the multicomponent analysis of CHP.
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Objective: To prepare pegylated long-circulating liposomes co-encapsulated by costunolide (Cos) and dehydrocostus lactone (DL), optimize the formulation and process, and evaluate the quality. Methods: The pegylated long-circulating liposomes co-encapsulated by Cos and DL were prepared by film hydration method. Single factor test and Box-Behnken response surface methodology were used to optimize the preparation process with encapsulation efficiency of Cos and DL as the index. The particle size, surface potential, encapsulation efficiency and in vitro release of the liposomes were evaluated. Results: The optimal preparation conditions were as follows: drug-to-lipid ratio was 0.14, ratio of cholesterol to phospholipid was 0.05, mPEG-2000-DSPE addition amount was 6%, hydration time was 30 min, and probe ultrasonic time was 4 min. The obtained liposome was round and uniform in distribution, with an average particle size of (104.8 ± 2.48) nm, a polydispersity index (PDI) of (0.245 ± 0.031), and a Zeta potential of (-9.7 ± 0.23) mV, the encapsulation efficiency of Cos and DL were (91.9 ± 2.6)% and (94.41 ± 1.23)%, respectively. Conclusion: The PEGylated long-circulating liposome prepared by the process and prescription optimization has good appearance and high encapsulation efficiency, which can meet the application requirements.
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Objective@#To develop a method of quantitative analysis of multi-components by single marker for nine effective constituents in Chenxiang-Huaqi tablets.@*Methods@#Using HPLC method, the mobile phase was acetonitrile -0.2% physcion acid in gradient elution program, the flow rate was 1.0 ml/min. The detection wavelength was set at 230 nm for liquiritin, ammonium glycyrrhizinate, costunolide and dehydrocostus lactone, and 283 nm for narirutin, naringin, hesperidin and neohesperidin, and 310 nm for pogostone. The column temperature was 30 ℃ and the injection volume was 10 μl. Using Hesperidin as the internal standard, the relative correction factors among liquiritin, ammonium glycyrrhizinate, costunolide, dehydrocostus lactone, narirutin, naringin, neohesperidin, pogostone were detected by QAMS and their contents were calculated. The content difference of the above nine components with the external standard method were compared.@*Results@#The calculated values of hesperidin, liquiritin, ammonium glycyrrhizinate, costunolide, dehydrocostus lactone, narirutin, naringin, neohesperidin, pogostone were not statistical significance from those measured by external standard method (P>0.05).@*Conclusions@#The method offered reference for quality standard of Chenxiang-Huaqi tablets.
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OBJECTIVE： To establish a method for simultaneous determination of matrine， oxymatrine， gallic acid， peoniflorin， costunolide and dehydrocostus lactone in Libiling tablets. METHODS： RP-HPLC method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with mobile phase consisted of methanol-0.1％ phosphoric acid （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelengths were 210 nm （matrine， oxymatrine） and 225 nm （gallic acid， peoniflorin， costunolide， dehydrocostus lactone）. The column temperature was set at 30 ℃， and sample size was 5 μL. RESULTS： The linear ranges of matrine， oxymatrine， gallic acid， peoniflorin， costunolide， dehydrocostus lactone were 0.053-5.28 mg/mL（r＝0.999 8）， 0.125-12.54 mg/mL（r＝0.999 9）， 0.013-1.33 mg/mL（r＝0.999 8）， 0.169-16.94 mg/mL（r＝0.999 9）， 0.048-4.77 mg/mL（r＝0.999 8）， 0.072-7.16 mg/mL （r＝0.999 9）. The limits of quantitation were 4.08×10－4， 4.48×10－4， 3.12×10－4， 2.10×10－4， 1.36×10－4， 1.84×10－4 mg/mL， respectively. The limits of detection were 1.24×10－4， 1.50×10－4， 1.02×10－4， 6.20×10－5， 4.20×10－5， 6.40×10－5 mg/mL， respectively. RSDs of precision， stability and reproducibility tests were all lower than 2％ （n＝6）. The recoveries were 98.03％-101.43％ （RSD＝1.25％， n＝6）， 97.73％-102.26％ （RSD＝1.96％， n＝6）， 97.18％-101.41％ （RSD＝1.98％，n＝6）， 97.45％-102.11％ （RSD＝1.88％，n＝6）， 96.85％-101.07％ （RSD＝1.75％， n＝6）， 97.12％-102.64％ （RSD＝1.82％，n＝6）， respectively. CONCLUSIONS： Established method is simple， stable and rapid， and can be used for simultaneous determination of 6 components in Libiling tablets.
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Objective To develop a method of quantitative analysis of multi-components by single marker for nine effective constituents in Chenxiang-Huaqi tablets. Methods Using HPLC method, the mobile phase was acetonitrile -0.2% physcion acid in gradient elution program, the flow rate was 1.0 ml/min. The detection wavelength was set at 230 nm for liquiritin, ammonium glycyrrhizinate, costunolide and dehydrocostus lactone, and 283 nm for narirutin, naringin, hesperidin and neohesperidin, and 310 nm for pogostone. The column temperature was 30 and the injection volume was 10 μl.℃ Using Hesperidin as the internal standard, the relative correction factors among liquiritin, ammonium glycyrrhizinate, costunolide, dehydrocostus lactone, narirutin, naringin, neohesperidin, pogostone were detected by QAMS and their contents were calculated. The content difference of the above nine components with the external standard method were compared. Results The calculated values of hesperidin, liquiritin, ammonium glycyrrhizinate, costunolide, dehydrocostus lactone, narirutin, naringin, neohesperidin, pogostone were not statistical significance from those measured by external standard method (P>0.05). Conclusions The method offered reference for quality standard of Chenxiang-Huaqi tablets.
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Objective:To establish a GC method for the imultaneous determination of cinnamaldehyde, bornyl acetate, costunol-ide,dehydrocostus lactone,magnolol and honokiol in Dutong pills. Methods: The determination was performed on an HP-5 column (30 m ×0.32 mm,0.25 μm) with programmed temperature. The carrier gas was nitrogen with the flow rate of 2.0 ml·min-1. The injection volumn was 1 μl and the sample split ratio was 5:1. The inlet temperature was 280 ℃. The detector was a flame ionization detector with temperature at 300 ℃. Results:The linear ranges were 32.28-516.40 μg·ml-1for cinnamaldehyde(r=0.999 3), 27.06-433.00 μg·ml-1for bornyl acetate(r=0.999 2),25.65-410.40 μg·ml-1for costunolide(r=0.999 3),26.10-417.60 μg ·ml-1for dehydrocostus lactone(r=0.999 3),24.01-384.20 μg·ml-1for magnolol(r=0.999 0) and 18.32-293.10 μg·ml-1 for honokiol(r=0.999 4). The average recovery was 99.71%(RSD=0.67%),99.34%(RSD=1.18%),100.16%(RSD=0. 34%),100.40%(RSD=0.39%),99.32%(RSD=1.22%) and 99.58%(RSD=0.58%)(n=6),respectively. Conclusion:The method is simple,sensitive and accurate,can be used to supplement the insufficient quality control of Dutong pills.
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Objective To comprehensively evaluate the Aucklandiae Radix quality by combining gray correlation method and FCM (Fuzzy C-Means) algorithm. Methods HPLC method was used to determine the content of costunolide and dehydrocostus, and the content of volatile oil was determined by steam distillation. The gray correlation method was used to sort the quality of Chinese herbal medicines, and the FCM algorithm was used to classify the quality of medicines. Results The average correlation degree of Aucklandiae Radix from different producing areas was: Yunnan > Guangdong > Guangxi > Hunan> Sichuan > Bozhou > Beijing > Hebei. FCM algorithm divided samples into three categories: Yunnan, Guangdong and Guangxi were in the high quality group; Hunan and Sichuan were in the medium quality group; Beijing, Bozhou, and Hebei were in the low quality group. Conclusion Constructing an integrated pattern recognition model system for evaluating the quality of Chinese medicinal materials, and the fuzzy clustering algorithm was adopted for the first time in Aucklandiae Radix. The purpose of this study is to form a kind of research method of Aucklandiae Radix quality evaluation and provide a way to guide and apply the new methods of modern pattern recognition and data mining in the application of traditional Chinese medicine quality evaluation.
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AIM To establish a quantitative analysis of multi-components by single-marker (QAMS) method for the simultaneous content determination of five constituents in Roudoukou-8 Powder (Myristicae Semen,Auck landiae Radix,Lignum aquilariae Resinatum,etc.).METHODS The analysis of 75％ methanol extract of this drug was performed on a 30 ℃ thermostatic Apollo C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of methanol-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 225,254,273,281 nm.With eugenol as an internal standard,the relative correction factors of the other four constituents were calculated,after which the content determination was made.RESULTS Ellagic acid,eugenol,costunolide,dehydroroma lactone,dehydrodiisoeugenol showed good linear relationships within the ranges of 0.227 0-1.135 2,5.272 2-26.361 0,0.540 8-2.704 0,0.530 4-2.652 0,0.059 0-0.299 5 μg (r ＞0.999 0),whose average recoveries (RSDs) were 96.37％ (2.07％),102.19％ (2.78％),101.66％ (1.66％),103.46％ (1.17％),98.25％ (1.98％),respectively.The results obtained by QAMS approximated those obtained by external standard method.CONCLUSION This stable and reliable method can be used for the quality control of Roudoukou-8 Powder.
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@# Objective: To investigate the effects of costunolide (Cos) on the proliferation, apoptosis, migration andinvasion of cholangiocarcinoma RBE cells, and explore its potential mechanism. Methods: The CCK-8, flow cytometry,Annexin V-FITC/PI double staining, Transwell assays were used to examine the influence of Cos on proliferation, cell cycle, apoptosis, migration and invasion of RBE cells after treated with gradient concentrations of Cos. The expressions of M M P 2 and M M P 9 were detected by qRT-PCR, and the expression of PI3K/AKT-associated signal proteins was detected by Western blotting. Results: Cosdose-dependently inhibited proliferation activity of RBE cells( P <0.05 or P <0.01), arrested cell cycle at S and G2/M phases and induced RBE cell apoptosis( P <0.01). Transwell and qRT-PCR results demonstrated that Cos impeded RBE cell migration, invasion, and reduced the transcription of M M P 2 and M M P 9 . Cos inhibited the expressionofp-AKT, Bcl-2, MMP2 and MMP9, the level of Bax. Conclusion: Cos restrained the proliferation, migration and invasion of RBE cells by suppressing PI3K/AKTpathway.
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Parthenolide is a sesquiterpene lactone derived from the plant feverfew (Tanacetum parthenium) that possesses multiple anti-inflammatory and anti-cancer properties. A specific, sensitive, accurate and precise LC-MS/MS method was developed and validated in the quantitative analysis of parthenolide in rat plasma. A liquid-liquid extraction method was used to separate the analyte and the internal standard (IS, costunolide) from plasma. A water-methanol mobile phase system was utilized in the gradient chromatographic separation. The calibration curve with good linearity (r2>0.99) was established between 2 and 128 ng·mL-1 with accuracy and precision within acceptable limits at different QC levels. High extraction recovery was achieved for both parthenolide (89.55%-95.79%) and IS (96.87%). Based on this LC-MS/MS method, the plasma stability and pharmacokinetics of parthenolide were assessed in rats. Parthenolide was proved to be very unstable in rat plasma, and was distributed and eliminated quickly in vivo, with a half-life less than 90 min. A high dose of parthenolide (80 mg·kg-1) resulted in a very low initial concentration (138.86±21.07 ng·mL-1). The systemic exposure of parthenolide (area under the curve) increased disproportionally from 40 mg·kg-1 dose group to 80 mg·kg-1 dose group. The present study may provide helpful information for the development of parthenolide as a drug candidate.
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Objective: To study the chemical constituents from fruits of Michelia yunnanensis. Methods: The chemical constituents were isolated and purified by column chromatographies on silica gel and Sephadex LH-20. Their structures were elucidated on the basis of spectroscopic data. Results: Twelve compounds were isolated from methanol extract of the fruits of M. yunnanensis and the structures were identified as 13-methoxy-11β, 13-dihydrocostunolide (1), lanuginolide (2), tulipinolide (3), lipiferolide (4), costunolide (5), parthenolide (6), 11β, 13-dihydroparthenolide (7), 4α, 5β-epoxy-13-methoxy-11βH-germacra-1 (10)-en-12, 6α-olide (8), cyperusol C (9), aromadendra-4β, 10α-diol (10), 9-oxonerolidol (11), and 11, 13-dehydrocompressanolide (12). Conclusion: Compound 1 is isolated as a new natural product. 1H-NMR and 13C-NMR data of compounds 1 and 2 are reported for the first time. Except compound 6, other compounds are isolated from this plant for the first time.
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Objective To develop an HPLC wavelength switching combined gradient elution method for simultaneous determi?nation of nine components in Kanglixin Jiaonang(KLXJN). Methods The analysis was performed on a Kromasil C18(4.6 mm×250 mm, 5μm)with gradient elution by using the mobile phase of acetonitrile-methanol(1:2)(A)-0.1%phosphoric acid solution(B). The col?umn temperature was maintained at 30℃and the flow rate was 0.9 ml/min. The detection wavelengths were set at 225 nm for costuno?lide(1)and dehydrocostus lactone(2),254 nm for aloe-emodin(3),rhein(4),emodin(5)and physcion(6),and 425 nm for bisde?methoxycurcumin(7),demethoxycurcumin(8)and curcumin(9). Results The calibration curves were linear within the range(μg/ml)of 6.610-132.2(r=0.9999)for 1,7.890-157.8(r=0.9996)for 2,14.07-281.4(r=0.9992)for 3,3.450-69.00(r=0.9997)for 4, 2.670-53.40(r=0.9998)for 5,3.760-75.20(r=0.9999)for 6,5.880-117.6(r=0.9996)for 7,8.490-169.8(r=0.9993)for 8,and 13.91-278.2(r=0.9991)for 9,respectively. The recoveries for 1,2,3,4,5,6,7,8 and 9 were 98.28%(RSD=1.09%),97.76%(RSD=0.80%),99.08%(RSD=1.72%),97.19%(RSD=1.00%),98.45%(RSD=1.24%),96.96%(RSD=1.21%),98.51%(RSD=1.55%), 97.52%(RSD=0.83%),and 100.04%(RSD=0.93%),respectively. Conclusion The established method is accurate,rapid and can be used for the quality control of KLXJN.
ABSTRACT
Objective:To analyze and evaluate the quality control of Radix Aucklandiae in Xianglian preparation and develop a new quality control method for the preparation through standard testing and exploration study on 127 batches of samples .Methods:Ac-cording to the mandatory method , Radix Aucklandiae was identified by TLC .In the exploratory research , the volatile compounds in Xianglian preparation was analyzed by SPME/GC-MS, and the GC-MS fingerprints of volatile compounds in Xianglian preparation with different dosage forms were established .The contents of costunolide and dehydrocostuslactone in Xianglian preparation were determined by UPLC.Results:According to the current quality standard , all the batches of samples met the standard with the qualified rate of 100%.The composition of volatile compounds in different dosage forms was similar , while the relative contents were quite different . The contents of costunolide and dehydrocostuslactone in Xianglian preparation were different .Conclusion: The exploratory research provides reference for the further revision of drug standard and monitoring quality of the preparation .