| Zizyphi Spinosi Semen(ZSS), the dried seeds of Ziziphus jujuba Mill. Var. Spinosa(Bunge) Hu ex H.F. Chou, has the function of medicine and foodstuff and has an excellent development prospect. Various experimental studies indicated that flavonoids from ZSS are the main bioactive compounds, and a further study of the flavonoids is pressing necessary. This study aims to prepare ZSS flavonoids-loaded solid lipid nanoparticles to enhance the biological availability. Furthermore, the study on metabolic kinetic characteristics and its metabonomics provides theoretical basis for further exploring of its pharmacodynamic action and mechanism.This topic research includes three parts:(1)The ZSS flavonoids-loaded solid lipid nanoparticles were prepared by the film-ultrasonication method in this experiment. Free drug was separated by Sephadex G-50 column from SLN and the ntrapment effieieney was used as the index for the optimization of the formulations. The response surface methodology was used to optimize the formulations of SLN. Response surface was delineated according to best-fit mathematic models, and the optimum formulation was selected by response surface. Quadratic multinomial models were better than multivariate linear models, and the regression coefficient of formulation was 0.9752. Bias between observed and predicted values both of optimum preparation technique and formulation were negligible, indicating the high predictability of the fit models.The SLN size formulated with the optimum techniques was485±2.8 nm met the quality specification of preparation. The model established by Expert-Design software is better to predict and could be used to optimize the preparation technique SLN by response surface methodology.(2)In this experiment, a HPLC-ESI-MS/MS method was developed and validated for the determination of 6’’’-feruloylspinosin and 6’’’-p-coumaroylspinosin in rat plasma with puerarin as the internal standard, and the rats were treated as the experiment objective by oral administration. The results showed that the method has good linear relationship,specificity, precision, stability and extraction recovery. The pharmacokinetics were gotten from the experiment and the blood drug concentration of 6’’’-feruloylspinosin and6’’’-p-coumaroylspinosin in the 45 min reached the maximum. Concentration-time cures had a index change and the semilog of the concentration had a linear relationship with time.There exists different change of AUC, Ke and T1/2 for 6’’’-feruloylspinosin and6’’’-p-coumaroylspinosin and eliminated the first order process in the body.(3)A metabonomic method based on reversed-phase liquid chromatography combining with tandem mass spectrometry(HPLC-Q-TOF/MS) has been established usingelectrospray ionization(ESI) in negative ion modes. Then this method combining with time-course analysis of discriminating metabolites was implemented to the metabonomic study on SD rats to find the potential biomarkers that were closely associated with anxiety and sedation. HPLC-Q-TOF/MS method had been established. HPLC-Q-TOF/MS in ESI negative methanol ion modes were used for metabonomics analysis of analyte, respectively.In addition, the precision, sensitivity and recovery rate of this method were investigated in detail using some representative blood metabolites and the results demonstrated that this method had high precision, good sensitivity and recovery rate, and could be used for the metabolomic analysis of rat blood. The blood metabonomic analysis of rats revealed 13 potential biomarkers that had regular change trend and were closely associated with anxiety and sedation, as 33 potential biomarkers in brain tissues metabonomic analysis, one of them were identified. |
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