ABSTRACT
The effect of intestinal flora changes on the pharmacokinetics of astragaloside Ⅳ in rats with type 2 diabetes mellitus was explored in this study. The rat model in preliminary experiment was established by high-sugar and high-fat diet combined with the intraperitoneal injection of low-dose streptozotocin(STZ). Rats were divided into model group, astragaloside Ⅳ group, berberine group and combination group(five rats in each group). After two weeks of gavage, the rats' feces was taken for 16 S rRNA sequencing of intestinal flora. Pharmacokinetic experiments were performed on astragaloside Ⅳ in the four groups one day after the preliminary experiment. Plasma samples were precipitated in methanol with ginsenoside Rb_1 as an internal standard, and the plasma concentrations of astragaloside Ⅳ at different time points were determined by UPLC-MS/MS. The chromatographic separation was performed on a Waters Acquity UPLC BEH-C_(18) column(2.1 mm×100 mm, 1.7 μm) via gradient elution. The mobile phase was acetonitrile(A) and 5 mmol·L~(-1) ammonium formate solution with 0.2% formic acid(B). The flow rate was 0.4 mL·min~(-1), the injection volume 5 μL and the column temperature 40 ℃. The mass spectrometry was carried out with electrospray ionization source(ESI) in multiple reaction monitoring and positive ion modes. The specificity, linearity range, accuracy, precision, stability and dilution effect of the method all met the requirements for the determination of astragaloside Ⅳ in plasma. Plasma concentration-time curves were plotted and relevant pharmacokinetic parameters were calculated by DAS 3.2.8. The results showed that the concentration of absorbed astragaloside Ⅳ increased within 0-3.95 h and began to decline since 3.95 h. After 36 h, the metabolism was complete. The area under the plasma concentration-time curve(AUC_(0-t)) and the peak concentration(C_(max)) of astragaloside Ⅳ were increased in the three administration groups compared with the model group, but without significant difference, which suggested that the pharmacokinetic characteristics of saponin components would not necessarily change after the drug-induced alteration of intestinal flora.
Subject(s)
Animals , Rats , Chromatography, High Pressure Liquid , Chromatography, Liquid , Diabetes Mellitus, Type 2 , Gastrointestinal Microbiome , Rats, Sprague-Dawley , Reproducibility of Results , Saponins , Tandem Mass Spectrometry , TriterpenesABSTRACT
Objective: To determine the content of index components in different parts of Gardenia jasminoides (pericarp, seeds, whiskers), study the fingerprint, and compare the contents and compositions differences of different parts of G. jasminoides, in order to provide the theoretical basis for different efficacies of G. jasminoides pericarp and seeds, explore the exploitation and utilization values of G. jasminoides whiskers, and avoid waste of gardenia medicinal resources. Method: The contents of geniposide and crocetin Ⅰ was were determined by HPLC, the content of total iridoid glycosides was determined by ultraviolet spectrophotometry, and three index components in different parts of G. jasminoides were analyzed. HPLC fingerprints of different parts of G. jasminoides were collected, the common pattern of HPLC fingerprints of different parts of G. jasminoides of different origins and with different processing methods was established, and the similarity evaluation software was used for data analysis; comparative analysis on fingerprints of different parts of G. jasminoides was conducted. Result: Content change of index components in G. jasminoides pericarp and seeds from Henan, Fujian and Jiangxi were the same. Content of geniposide:Fujian > Henan > Jiangxi, the contents of three components in G. jasminoides pericarp from Fujian were much higher than those from Henan and Jiangxi, the contents of crocetin Ⅰ and total iridoid glycosides:Fujian > Jiangxi > Henan, the contents of total iridoid glycosides from Fujian, Jiangxi were much higher than those from Henan. The order of three index components in G. jasminoides whiskers from different origins from high to low, the content of geniposide and crocetin Ⅰ was Fujian > Jiangxi and Henan, the content of total iridoid glycosides was Fujian > Jiangxi > Henan.In the same part, there were 22 common peaks in the fingerprints of G. jasminoides pericarp, except for S13-S15, the similarity of other samples were more than 0.9;the fingerprints of G. jasminoides seeds had 22 common peaks, except for S22-S30, the similarities of other samples were more than 0.9;the fingerprints of G. jasminoides whiskers had 16 common peaks, except for S7-S9, the similarities of other samples were more than 0.9.In different parts, the fingerprints of G. jasminoides whiskers were significant different from those of pericarp and seeds, the number of peaks in G. jasminoides whiskers reduced, the order of height of peaks 2, 3, 5 of G. jasminoides from high to low were whiskers > gardenia > seeds. There was not peak X in the seeds, the height of peak X of gardenia in whiskers was higher than that in pericarp, except for the peak 17, the height of all peaks in seeds were higher than that in whiskers. Conclusion: There are significant differences in the contents of index components in G. jasminoides pericarp and seeds. The content of total glycosides in gardenia is high, suggesting that it can be used to extract total iridoid glycosides. The fingerprints can reflect the content difference and species distribution of different parts of G. jasminoides, so as to provide theoretical support for the studies for pharmacodynamic material basis of G. jasminoides and the scientificity and rationality of the separate application of G. jasminoides pericarp and seeds.
ABSTRACT
Objective To optimize the extraction and alcohol precipitation process of Qizhi Yifei Granules by multi index orthogonal experiment. Methods With extraction rate of astragaloside in Astragali Radix, quercetin-3-O-β-D-glucose-7-O-β-D-gentian diglucoside in Descurainiae Semen Lepidii Semen and yield rate of dry extract as indexes, the extraction process of Qizhi Yifei Granules was optimized. Taking the retention rate of astragaloside and quercetin-3-O-β-D-glucose-7-O-β-D-gentian diglucoside as indexes, the alcohol precipitation process was optimized. Results The best water extraction process was as follows: adding 10 times amount of water, extracting for 1.5 h, 3 times. The optimum alcohol precipitation process was: concentrated to the relative density of 1.05–1.10 (60 ℃), adding ethanol to 60％ and alcohol precipitation. Conclusion The optimized extraction and alcohol precipitation process is stable and feasible, which can provide the basis for the preparation.
ABSTRACT
To establish HPLC-MS/MS method for simultaneous determination of daphnetin, daphnoretin, and daphneticin in rat plasma after oral and intravenous administration of Daphne giraldii extract, and then use them in the calculation of pharmacokinetic parameters. Six sprague-dawley rats received intragastric administration of D. giraldii extract (daphnetin, daphnoretin and daphneticin were 88.40, 3.24 and 4.28 mg•kg⁻¹, respectively). Their drug plasma concentration was determined by LC-MS/MS with schisandrin as an internal standard to draw plasma concentration-time curve. The pharmacokinetic parameters were calculated by Kinetica 4.4. The results showed that the linear range was 5-1 000 μg•L⁻¹ for daphnetin, daphnoretin and daphneticin, and the method ological test showed conformance to the requirements.The intraday and inter-day variable coefficients (RSD) were both less than 15.0%, indicating that both of legitimate precise and accuracy were consistent with the analysis requirements of biological samples. For daphnetin, the pharmacokinetic parameters Tmax, Cmax, AUC0-t, T1/2 and MRT were 4 h, 858.96 μg•L⁻¹, 10 566.4 μg•L⁻¹•h, 5.19 h and 9.43 h, respectively. For daphnoretin, the pharmacokinetic parameters Tmax, Cmax, AUC0-t, T1/2 and MRT were 2.92 h, 178.00 μg•L⁻¹, 905.89 μg•L⁻¹•h, 3.50 h and 6.95 h, respectively. For daphneticin, the pharmacokinetic parameters Tmax, Cmax, AUC0-t, T1/2 and MRT were 2 h, 36.67 μg•L⁻¹, 355.11 μg•L⁻¹•h, 4.95 h and 8.27 h, respectively. The LC-MS/MS analysis method established in this study was proved to be so accurate and sensitive that it can be applied to the pharmacokinetic study of daphnetin, daphnoretin and daphneticin.
ABSTRACT
Ischemic brain injury is a major disease which threatens human health and safety. (3, 5, 6-trimethylpyrazin-2-yl) methyl 3-methoxy-4-[(3, 5, 6-trimethylpyrazin-2-yl) methoxy] benzoate (VA-T), a newly discovered lead compound, is effective for the treatment of ischemic brain injury and its sequelae. But the poor solubility of VA-T leads to poor dissolution and limited clinical application. In order to improve the dissolution of VA-T, the pharmaceutical technology of solid dispersions was used in the present study. VA-T/polyvinylpyrrolidone (PVP) solid dispersion was prepared by the solvent method. The dissolution studies were carried out and solid state characterization was evaluated by differential scanning calorimetry (DSC), infrared spectroscopy (IR), x-ray diffraction (XRD) and scanning electron microscopy (SEM). The dissolution rate of VA-T was significantly improved by solid dispersion compared to that of the pure drug and physical mixture. The results of DSC and XRD indicated that the VA-T solid dispersion was amorphous. The IR spectra showed the possible interaction between VA-T and PVP was the formulation of hydrogen bonding. The SEM analysis demonstrated that there was no VA-T crystal observed in the solid dispersions. The ideal drug-to-PVP ratio was 1:5. In conclusion, the solid dispersion technique can be successfully used for the improvement of the dissolution profile of VA-T.