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This study aims to establish a method for the determination of the concentration of five main components of phthalide target areas of Chaxiong(CPTA) and its inclusion of β-CD in the plasma of rats, and determine the pharmacokinetic parameters, absolute bioavailability and relative bioavailability of CPTA/β-CD inclusion compound in vivo. The plasma concentrations of senkyunolide A, N-butylphthalide, new osthol lactone, Z-ligustilide and butenyl phthalide were determined with UPLC-MS/MS. The content determination was conducted at the chromatographic conditions as follows: Shim-pack GIST C_(18)-AQ HP column(2.1 mm×100 mm, 3 μm), mobile phase of 0.1% formic acid solution(A)-acetonitrile(B), gradient elution, flow rate of 0.3 mL·min~(-1), column temperature of 35 ℃ and injection volume of 2 μL. The mass spectra were obtained with electrospray ion source(ESI), positive ion mode and multi reaction monitoring. CPTA/β-CD inclusion compound was prepared by grinding method, DAS 2.0 software was used to model the data, and the absolute bioavailability of CPTA and relative bioavailability of inclusion compound were calculated. Finally, the methods for the determination of five components of senkyunolide A, N-butylphthalide, new osthol lactone, Z-ligustilide and butenyl phthalide in CPTA, were successfully established. The linear relationship among the five components was good within their respective ranges, r>0.99. The absolute bioavailability of the five components in rats was 22.30%, 16.32%, 21.90%, 10.16% and 12.43%, respectively. After CPTA/β-CD inclusion was prepared, the relative bioavailability of the five components was 138.69%, 198.39%, 218.01%, 224.54% and 363.55%, respectively, significantly improved. This method is rapid, accurate and sensitive, so it is suitable for the pharmacokinetic study of extracts in traditional Chinese medicine and their preparations.
Subject(s)
Animals , Rats , Benzofurans , Chromatography, High Pressure Liquid , Chromatography, Liquid , Rats, Sprague-Dawley , Reproducibility of Results , Tandem Mass SpectrometryABSTRACT
Objective: To evaluate the effect of manual decocting and machine decocting on the chemical constituents in Bazhentang based on non-targeted metabolomics, and to find the differential chemical constituents of these two decocting methods. Method: Bazhentang was boiled by standardized manual decocting and machine decocting methods,respectively. Orthogonal partial least square-discriminate analysis(OPLS-DA) and other multivariate statistical methods, combined with variable importance in the projection(VIP) value and t-test, were employed to analyze the effect of two decocting methods on the chemical constituents in Bazhentang. The differential chemical constituents were analyzed by UPLC-LTQ-Orbitrap MS under positive and negative ion modes,mobile phase was acetonitrile-0.1%formic acid aqueous solution for gradient elution,the scanning range was m/z 50-1 500. Result: Under the positive and negative ion modes of high-resolution mass spectrometry, a total of 87 differential components were found,40 of them were identified according to the mass spectrometry data and literature reports, including senkyunolide A, glycyrrhizin, ferulic acid, etc. Conclusion: Based on the analysis of color and chemical compositions of Bazhentang, there are obvious differences between the standardized manual decocting and machine decocting. If the advantages of these two methods are combined,a standardized decoction process can be established on the basis of maintaining the advantages of manual decocting, the effectiveness of traditional Chinese medicine decoction will be maximized and it will be convenient for patients to take it.
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Objective: To optimize the processing technology for prepared Chuanxiong Rhizoma by D-optimal response surface methodology with comprehensive weighted of multi-index. Methods: The single factor experiment was used to optimize the dilution ratio and the moistened time of rice wine. Then, based on the contents of five main components (ferulic acid, ligustilide, senkyunolide A, senkyunolide I, and 3-butylidenephthalide), the D-optimal response surface methodology was used to optimize the rice wine dosage, stir-fried temperature, and stir-fried time in the processing technology for prepared Chuanxiong Rhizoma. Results: The optimal processing technology for prepared Chuanxiong Rhizoma was mixed with 20% rice wine, moistened for 75 min, and stir-fried for 12 min at 156 ℃. Also, it was found that the contents of senkyunolide A, senkyunolide I, and 3-butylidenephthalide in prepared Chuanxiong Rhizoma had increased, and it should be associated with processing. Conclusion: The processing technology for prepared Chuanxiong Rhizoma optimized by D-optimal response surface methodology with comprehensive weighted of multi-index can be used to optimize the process parameters, promote the stability and repeatability of the processing technology, which can significantly control the inner quality of prepared Chuanxiong Rhizoma to ensure effectiveness in clinical application.
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Objective The main metabolites of Yaobitong Capsule (YC) in rat urine and bile were investigated by UPLC-Q-TOF- MS/MS, and their major metabolic pathways were summarized. Methods To compare with the control samples, the possible metabolites were found. Their structures were speculated by exact mass and fragment ions. Results YC had 31 metabolites in urine and 35 metabolites in bile under positive and negative ion mode. They were mainly the metabolites of ligustilide, 3-butyl-phthalide, osthole, senkyunolide A, sankyunolide A, emodin, decursinol/marmesin/columbianetin, corypalmine/ isocorypalmine, corybulbine/isocorybulbine/coryphenanthrine, allocryptopine, ferulic acid, cryptochlorogenic acid, angelol and so on. Their major metabolic pathways included hydrolysis of the lactone ring, hydroxylation, demethylation, dehydration, dehydrogenation, dehydroxylation, hydrogenation, methylation, methyl oxidation, glucuronidation, sulphate-binding, N-acetylcysteine-binding, cysteine-binding, cysteinyl acid-glycine-binding, glutathione-binding and so on. In addition, three prototype components, namely paeoniflorin, albiflorin, and ginsenosides Rg1, were detected in rats’ bile. Conclusion The established UPLC-Q-TOF-MS/MS method can analyze the metabolites of YC capsule in rat urine and bile. The in vivo metabolic characteristics of YC have been initially clarified. The results provide a foundation for elucidating the in vivo efficacy ingredients of YC.
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OBJECTIVE: To study the influence and mechanism of bioactive ingredients of Ligusticum chuanxiong Hort. on the transport of gastrodin based on cell culture model in vitro. METHODS: Cell toxicity of gastrodin, ligustilide, senkyunolide I and senkyunolide A were detected by MTT assay. The transport mechanism of gastrodin and the influence of ligustilide, senkyunolide I and senkyunolide A on the transport of gastrodin were studied in MDCK-MDR1 monolayer cells. The changed expressions of P-gp caused by ligustilide, senkyunolide I and senkyunolide A were analyzed by Western blotting. RESULTS: Gastrodin showed relatively poor absorption in MDCK-MDR1 cells for its apparent permeability coefficients were less than 1×10-6 cm·s-1. P-gp inhibitor made the Papp(B→A)/Papp(A→B) of gastrodin reduced from 1.29 to 0.79, which indicated that the transport of gastrodin was influenced by P-gp. In the presence of ligustilide(30 μg·mL-1) or senkyunolide I(120 μg·mL-1), the Papp(A→B) of gastrodin in MDCK-MDR1 were significantly increased(P<0.01). In the presence of senkyunolide A(120 μg·mL-1), the Papp(A→B) of gastrodin in MDCK-MDR1 were markedly increased(P<0.05). High, medium and low dose of ligustilide, senkyunolide A and senkyunolide I could significantly inhibit the expression of P-gp protein. CONCLUSION: The results indicate that the transport mechanism of gastrodin might be passive diffusion as the dominating process with the active transportation mediated mechanism involved. Ligustilide, senkyunolide A and senkyunolide I increase the transport of gastrodin attribute to down-regulate P-gp expression.
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Objective To study the pharmacokinetic features of ferulaic acid, senkyunolide A and ligustilide in Buyang Huanwu associated prescriptions (Buyang HuanwuDecoction andNaojianTablets).MethodsHPLC-DAD was applied for simultaneous determination plasma concentration of three ingredients with jugular venous cannula rats after intragastric administration ofBuyang Huanwu associated prescriptions. The pharmacolinetic parameters of each ingredient was calculated by DAS2.0, and then the total quantum statistical moment (TQSM) standard similarity was used to measure the overall pharmacokinetics behaviors.Results There were great differences in the three ingredients after the administration of two prescriptions, while the total quantum statistical parameters were very closely. The TQSM pharmacokinetic parameters of the three components inBuyang HuanwuDecoction andNaojian Tablets showed that AUC, MRT, VRT were 240.6 and 133.0, 3.192 min and 3.259 min, 21.59 min2and 19.75 min2, respectively.The similarity was up to 0.977 8.Conclusion The metabolic processes in vivo ofBuyang Huanwu Decoction andNaojianTablets have similarities. The efficacy of Chinese herbal compounds mostly depends on the multi-components overall contributions.
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Objective: To investigate the pharmacokinetics of ligustilide, senkyunolide A, and butylphthalide in supercritical CO2 extraction (SFE) fraction of Xiongqiong Decoction after ig administration in rats. Methods: Plasma samples were collected according to different time periods after ig administration, and the concentration of ligustilide, senkyunolide A, and butylphthalide in plasma samples was determined by HPLC so as to calculate the pharmacokinetic parameters and establish the metabolism curves by PKSolver 2.0 pharmacokinetic software. Results: Excellent liner relationship of ligustilide, senkyunolide A, and butylphthalide was obtained in the range of 0.416-16.640 μg/mL, 0.40-16.05 μg/mL, and 0.52-31.50 μg/mL, respectively. The recovery of the method was more than 84%, and the intra- and inter-day RSD values were less than 5%. The pharmacokinetics parameters of ligustilide were as follows: t1/2 (2.082 0 ± 0.637 6) h, Cmax (3.272 5 ± 0.955 5) μg/mL, AUC0-t (15.400 ± 1.812) μg∙h/mL, and the pharmacokinetics parameters of senkyunolide A and butylphthalide were similar to those of ligustilide. Conclusion: The method for the determination of three effective components from Xiongqiong Decoction in plasma samples is precise, specific, and suitable for the pharmacokinetic study and determination of the three effective components simultaneously.
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Objective: To study the dissolution characteristics of Qishe Pills in two kinds of media and to lay the foundation for the establishment of quality evaluation methods by stripping method. Methods: With 1% Polysorbate 80 solution and 0.5% sodium dodecyl sulfate (SDS) as media, the sinomenine, calycosin-7-glucoside, senkyunolide I, ononin, calycosin, formononetin, and senkyunolide A, determined by HPLC, were used as indexes. The cumulative dissolution of Qishe Pills at different time was investigated, and the dissolvable situation of respective components in the different media was compared simultaneously. The dissolution models were fit and the dissolution parameters were obtained. Results: The linear ranges of sinomenine, calycosin-7-glucoside, senkyunolide I, ononin, calycosin, formononetin, and senkyunolide A were 0.718-10.770, 0.190-2.850, 0.100-1.500 μg, 31.4-471.0, 34.0-510.0, 33.6-504.0, and 40.0-600.0 ng. In 45 min, the cumulative dissolving rates of the respective components were above 90% (calycosin-7-glucoside excepted) with 1% Polysorbate 80 solution as medium, while they were slower with 0.5% SDS as medium. The Weibull model of the seven components was good in the two kinds of media, while other model of the individual components had higher correlation in the different media. The f2 similarity factor method indicated that the dissolution of the same component of Qishe Pills has no similarity in the two different media. Conclusion: The dissolvable speed of each component with 1% Polysorbate 80 as medium was faster than that with 0.5% SDS as medium obviously. With 1% Polysorbate 80 as medium, the dissolution of each component had similarity and synchronism. Using 1% Polysorbate 80 solution as medium is applicable to research the dissolution of Qishe Pills.
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AIM: To research how to separate the active component in Ligusticum chuanxiong Hort by high speed counter current chromatography. METHODS: Senkyunolide A and Z-ligustilide,the main components of volatile oil in Ligusticum chuanxiong Hort were one step separated by high speed counter current chromatography. n-hexane-ethyl acetate-ethanol-water,1 ∶ 1 ∶ 1 ∶ 1(v/v),was used as the solvent system for HSCCC. Top phase and bottom phase were respectively used as static phase and mobile phase. Optimum speed and flow rate were 900 r/min and 1. 2 mL/min respectively. RESULTS: Collected fractions were analyzed by HPLC and identified by EI-MS and 1HNMR. Purity could reach more than 95% . CONCLUSION: Lactone is fit to be separated and prepared by high speed counter current chromatography with good resolution and high purity. We find a fast and efficient way to separate these.