Hydrogen To Eritrea, Phenol In Mice Of Pharmacokinetic Process In Vitro And In Vivo Studies

Common rush, which is the dried medulla of Juncus effuses L., has been used to eliminate excess heart fire and to induce diuresis for treatment of restlessness and insomnia etc. Dehydroeffusol, which is the major phenanthrene in Juncus effusus, has been conformed to exhibit potent anxiolytic effect by our previous study. Our group has tested a serious of analogues and synthetic compounds which are derived from dehydroeffusol. Pharmacodynamic research proved that some compounds have potent anxiolytic effect. To develop dehydroeffusol and its analogues as new anxiolytics, a system pre-clinic pharmacokinetic study of dehydroeffusol in mice was performed as follows:1. Literature review sectionThis section includes the common research methods of herbal pharmacokinetics, pretreatment methods of biological sample analysis, application of LC-MS in herbal pharmacokinetic, effects of phenanthrenes on the central nervous system and the research progress of phenanthrenes in vivo metabolism.2. The development of quantification method for the determination of dehydroeffusol in biological samples.A sensitive and specific LC-MS method was developed and validated for the quantification of dehydroeffusol in biological samples of mice. A simple liquid-liquid extraction method with ethyl acetate was used to treat the biological samples, then the samples were saperated on an Agilent HC-C18column by using methanol-water (76:24) as mobile phase. Electrospray ionization (ESI) source was applied and operated in negative ion modle; SRM scan. The precision and accuracy of the method were acceptable for bio-analytic analysis.The method has been successfully applied to the pharmacokinetic study of dehydroeffusol in mice.3. The absorption pharmacokinetics of dehydroeffusol in miceSingle oral doses of5,10, and20mg·kg-1of dehydroeffusol were given to mice respectively. LC-MS method was used for the quantification of dehydroeffusol in mice plasma samples in different time. Plasma concentration-time data were analyzed by non-compartmental model with WinNonlin4.1. The results showed that after single oral dose of dehydroeffusol, the pharmacokinetics of dehydroeffusol was fitted to rapid obsorption, rapid distribution and slow elimination and was eliminated as first-order kinetics. The t1/2value were260.9、204.4、213.9min respectively; The Cmax value were12.27、27.94、60.30ng·mL-1respectively; The AUC value were545.7、2105.2、3013.1min*ng·L-1respectively. A nonliner relationship was showed between the AUC and dose.4. Tissue distribution of dehydroeffusol in miceSingle oral doses of5mg·kg-1of dehydroeffusol were given to mice. LC-MS method was used for the quantification of dehydroeffusol in mice tissue samples in different time. Tissue concentration-time data were analyzed by non-compartmental model with WinNonlin4.1. The results showed that the tissue drug concentrations of liver> kidney> brain, were all higher than the plasma drug concentration, suggesting that the high rate affinity of dehydroeffusol with tissue. Dehydroeffusol can be rapidly absorbed through the blood-brain barrier into the brain tissue, and eliminated slowly in brain, which suggested brain may be the target organ for its anti-anxiety effect.5. Metabolism of dehydroeffusol in miceLC-ESI-MS assay was used to investigate the metabolites in urine and feces. And a total of19metabolites were found. The result showed that dehydroeffusol exist in vivo mainly as its phase H metabolites including aglycone glucuronides, sulfates and their mixtures.