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Objective:To observe the effect of Chaibei Zhixian decoction and peimine on Carbamazepine (CBZ) concentration, P-glycoprotein (P-gp) and multi drug resistance 1(MDR1) expression in the brain tissues of rats with refractory epilepsy, and to understand the contribution of Peimine in the compound prescription to treat the refractory epilepsy. Method:Epilepsy rat models were established by injecting kainic acid (KA) in the lateral ventricle. The successfully modeled rats were randomly divided into model group, CBZ group(0.12 g·kg-1),Chaibei Zhixian decoction+CBZ group(8.39 g·kg-1+0.12 g·kg-1), peimine+CBZ group(0.01 g·kg-1+0.12 g·kg-1) and sham operation group. After 60 days of intervention, the expression levels of P-glycoprotein (P-gp) and MDR1b mRNA in the brain cortex were detected by Western blot and quantitative real\|time fluorescence polymerase chain reaction(Real-time PCR),the contents of CBZ and 10,11-epoxidation of carbamazepine (CBZE) were measured by liquid chromatography-mass spectrometry (LC-MS). Result:Compared with sham group, the expression of P-gp/MDR1 in the cortex of model group was significantly increased (PPPPPPPConclusion:Chaibei Zhixian decoction and peimine may increase the content of CBZ and CBZE in the brain tissues in rats with intractable epilepsy by reducing the expression of MDR1/P-gp in the cortex.
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Objective: To develop an HPLC-ELSD method for the determination of amygdalin, aucubin, harpagide, peimisine, peimine and peiminine in Keling capsules simultaneously.Methods: An Ultimate XB C 18 (250 mm× 4.6 mm , 5 μm) chromatographic column was adopted with an ELSD (the drift tube temperature was 105℃, the flow rate of nitrogen was 2.0 L·min-1).The mobile phase was acetonitrile-methanol (1∶1) and 0.4% acetic acid solution with gradient elution at a flow rate of 0.7 ml·min-1 , and the column temperature was set at 35 ℃.Results: Amygdalin, aucubin, harpagide, peimisine, peimine and peiminine was linear within the range of 13.56-271.20 μg·ml-1 (r=0.999 2), 8.48-169.60 μg·ml-1 (r=0.999 9), 4.89-97.80 μg·ml-1 (r=0.999 7), 2.66-53.20 μg·ml-1 (r=0.999 4), 1.82-36.40 μg·ml-1 (r=0.999 8) and 2.04-40.80 μg·ml-1 (r=0.999 6), respectively.The average recovery and the corresponding RSD were 97.90% (1.20%), 99.21% (1.62%), 97.68% (0.75%), 98.36% (1.38%), 99.70% (0.79%) and 97.95% (1.56%)(n =6), respectively.Conclusion: The method is simple and specific, and the results are accurate and repeatable.The method is helpful to the quality control of Keling capsules.
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Objective: To study the influence of sulfur-fumigation on pharmacokinetic of peimine and peiminine of Fritillaria thunbergii in rat plasma by LC-MS/MS. Methods: After random grouping, 18 SD rats were given the solution of fresh-cut and sulfur-fumigated F. thunbergii by ig administration. The blood drug concentration of peimine and peiminine in rat plasma was determined by HPLC-MS/MS, and the pharmacokinetic parameters were calculated with 3P97 software. Results: The pharmacokinetic parameters of peimine and peiminine of sulfur-fumigated F. thunbergii in rat plasma were (66.40 ± 4.65), (146.72 ± 10.88) ng/mL for Cmax, and (181.79 ± 7.85), (457.38 ± 58.81) ng∙h/mL for AUC, respectively. Those of fresh-cut sample in rat plasma were (186.37 ± 18.8), (227.65 ± 7.01) ng/mL for Cmax, and (197.70 ± 18.69), (566.16 ± 41.55) ng∙h/mL for AUC, respectively. The pharmacokinetic parameters of perimine and peiminine of sulfur-fumigated sample in rat plasma were less than those of fresh-cut sample. Conclusion: The results showed that sulfur-fumigation decreased the bioavailability of peimine and peiminine. This study could provide a basis for further clarifying the influence of sulfur-fumigation on efficacy material base of F.thunbergii.
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Objective: To develop an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of six components (indirubin, peimine, peiminine, ginkalide A, ginkalide B, and ginkalide C) in Baikening Granules. Methods: Preparing the samples solution by Soxhlet's extraction using methanol-chloroform as solvent, and multiple reaction monitoring (MRM) with a polarity-switching electrospray ionization (ESI) source between positive and negative modes was used for the quantification of indirubin, two peimisines, and three bilobalides. The detection was performed on an Inertsutain C18 column (75 mm × 3.0 mm, 2 μm) using a mobile phase consisted of methanol-acetonitrile and 2 mmol/L ammonium acetate water solution with gradient elution lasting 6 min. Results: All of the analytes showed good linearity (r ≥ 0.995 8) in the tested ranges. The precision, repeatability, and stability of the method were good for the six components. The average recoveries were in the range of 96.5%-105.1% with relative standard deviations (RSD) ≤ 4.2%. Conclusion: The new established polarity-switching LC-MS-MS method has been proven to be highly sensitive and effective in evaluating the quality of Baikening Granules, and peimine, peiminine, ginkalide A, ginkalide B, and ginkalide C are quantified in this drug for the first time.
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Objective: To identify and analyze the chemical constituents in Yinhuang Qingfei Capsule by HPLC-Q-TOF-MS/MS method. Methods: The HPLC method was used with the conditions that the column was Inertsil ODS-2 C18 (250 mm × 4.6 mm, 5 μm). Columu and electrospray ion (ESI) source was employed for the qualitative analysis under positive ion mode. These components were further analyzed by MS spectra, and by comparing with the corresponding reference substances and literature data. Results: According to the MS principle and literature data, 54 compounds were identified from the sample of Yinhuang Qingfei Capsule. Conclusion: An efficient HPLC-Q-TOF-MS/MS approach has been established for studying the chemical constituents in Yinhuang Qingfei Capsule, which paves a way for the quality control and further substance basis studies of the preparation.
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Objective To develop a new high performance liquid chromatography (HPLC ) coupled with Evaporative Light Scattering Detector (ELSD) method for simultaneous determination of two indicative components (peimine and peimi-nine) in Xiaoer Baotaikang granules .Methods The analysis was performed on a Thermo C18 column (250 mm × 4 .6 mm , 5 μm) by using a mobile phase consisted of acetonitrile-water and diethylamine (65 ∶ 35 ∶ 0 .03) ,and the flow rate was 1 .0 ml/min;ELSD parameters :the temperature of drift tube was 85 ℃ and the carrier gas flow rate was 2 .2 L/min .Results The linear ranges of peimine and peiminine were 0 .586~5 .860 μg (r=0 .999 6) and 0 .564~5 .640 μg (r=0 .999 9) ,respec-tively .The average recovery rate for peimine and peiminine was 99 .44% (RSD was 0 .7% ) and 98 .56% (RSD was 1 .0% ) , respectively .Conclusions The method is sample ,accurate and reproducible ,which could be used for the quality control of Xiao-er Bao taikang g ranules .
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Fritillaria thunbergii Miq. has been widely used in traditional Chinese medicine for its expectorant, antitussive, antiinflammatory and analgesic properties. Moreover, modern pharmacological studies have demonstrated that F. thunbergii Miq. has efficacy in the treatment of leukemia and cancers of the liver and cervix. Although the alkaloid, peimine, is largely responsible for these pharmacological effects, it has very low oral bioavailability. The aim of this study was to investigate the intestinal absorption of peimine in Caco-2 cell monolayers. Having demonstrated that peimine is non-toxic to Caco-2 cells at concentrations <200 μmol/L, the effect of peimine concentration, pH, temperature, efflux transport protein inhibitors and EDTA-Na2 on peimine transport were studied. The results show that peimine transport is concentration-dependent; that at pH 6.0 and 7.4, the P app(AP-BL) of peimine is not significantly different but the P app(BL-AP)) is; that both P app(AP-BL) and P app(BL-AP) at 4 °C are significantly higher than their corresponding values at 37 °C; that the P-glycoprotein (P-gp) inhibitors, verapamil and cyclosporin A, increase absorption of peimine; and that EDTA-Na2 has no discernible effect. In summary, the results demonstrate that the intestinal absorption of peimine across Caco-2 cell monolayers involves active transport and that peimine is a substrate of P-gp.
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To establish an optimized determination method for the alkaloids peimine and peiminine in three varieties of Fritillaria. Methods:The samples were refluxed in a mixed solution, the pH value of the mixed solution was adjusted, and then the solution was extracted by ether to obtain the test solution. A Shim-pack VP-ODS column (4. 6 mm × 250 mm,5 μm) was used, the mobile phase consisted of acetonitrile-0. 02% diethylamine(75∶25), and the flow rate was 1. 0 ml·min-1. An Alltech 2000ES ELSD detector was used, the drift tube temperature and the flow rate of nitrogen carrier gas in the ELSD was set at 93℃ and 2. 4 L·min-1 , respectively. The regression equation was calculated between the logarithm values of the reference solution concentration and the corre-sponding peak area. Results:The method could effectively extract the alkaloids from the samples. Peimine and peiminine were well separated by the method, and showed good linearity within the respectively given ranges. The average recovery of the alkaloids in the three varieties of Fritillaria was within the range of 95. 22%-103. 51%. Conclusion: The method is simple,convenient and universal with easy operation,which provides references for the determination of alkaloids in the other varieties of Fritillaria.
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Objective: To simultaneously determine the contents of five components (amygdalin, paeoniflorin, peimine, astragaloside IV and rutaecarpine) in Fuzheng Pingxiao Capsula by LC-MS. Methods: Chromatographic separation was achieved with gradient elution by Agilent Eclipse plus C18 column (250 mm x 4.6 mm, 5 μm) and an Agilent 1100 Mass Spectrometer system was operated under the SIM mode with electrospray positive ionization (ESI). The mobile phase is acetonitrile-0.1% methanoic acid. Results: The LOQ of amygdalin, paeoniflorin, peimine, astragaloside IV, and rutaecarpine in Fuzheng Pingxiao Capsula were 12.9, 32.2, 1.00, 1.21, and 0.40 ng/mL, and the LOD were 6.46, 6.44, 0.25, 0.61, and 0.16 ng/mL, respectively. Within the linear range, r > 0.999 0. Both intra-day and inter-day precision with RSD was less than 2%. The average recovery rates of the five components were in the range of 98%-102%. Conclusion: This method is fast, sensitive, and reproducible. It could be used to determine amygdalin, paeoniflorin, peimine, astragaloside IV, and rutaecarpine in Fuzheng Pingxiao Capsula under the same chromatogram condition.
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AIM: To promote the quality standard of Senbei Beigua Oral Thick Paste (Fructus Cucurbitae, Radix Codonopsis, Radix Adepophorae, Bulbus Fritillariae Thunbergii, Rhizoma Zingiberis Recens) . METHODS: Rhizoma Zingiberis Recens, Fructus Cucurbitae were identified by TLC. Peimine content was determined by TLCS. RESULTS: The study showed that spots of samples on TLC can be well separated and the method had strong specificity. The quantification method had a good linear at the range of 0.326-1.194 ?g. The average recovery was 96.8% and RSD was 0.87%. CONCLUSION: The methods for identification and quantification are simple, accurate and reproducible. It can be used for quality control of Senbeibeigua Oral Thick Paste.