Analysis Of Blood Components Of Compound Anti-anxiety Capsule Based On Serum Pharmacology And In Vivo Metabolism Study Of Eugenol Glucoside

Compound Anti-anxiety Capsules(CAC)is a formula treating anxiety related diseases in clinic,which is composed of Valerianae Jatamansi Rhizoma,Albizia Cortex,the parched Ziziphi Spinosae Semen and Junci Medulla.The function of CAC is regulating vital energy,nourishing blood and quieting the spirit.Clinic observation and pharmacology experiments show that CAC have remarkable anti-anxiety effect.In previous studies,the quality standard,the anxiolytic mechanism and the safety of CAC had been studied systematically.In order to clarify the material basis and compatibility mechanism of CAC,and founded for traditional Chinese medicine with high bioavailability,safety and reliable efficacy,UPLC-Q-Tof-MS/MS was using to analysis the component of CAC in vitro and in the serum to set a reliable analytical method,and provide a complete database for serum drug chemistry and to optimize the analysis condition of the component in blood.Furthermore,UPLC-TQMS/MS was using to establish method for determining contents of SAG in biological sample,and pharmacokinetics of SAG and its aglycone SYR was fully studies on this research.The main research contents and archieved results are as follow:1.Study on Pharmacodynamic material basis of CACUnder the positive/negative ion mode,to analysis the component of CAC,CAC without single material,and the medicated serum of rats,a stable condition of UPLC-Q-Tof-MS/MS was established.The cracking behaviors of standard samples in CAC were illuminated,particularly the cracking behaviors of Parallel tetrahydrofuran lignin.The components of four medicinal materials which influence the spectrum peak were found out by PCA and OPLS-DA,and the components was identified on the basis of mass spectra fragmentation,the cracking behaviors of standard sample,and related researches on Valeriana,Jujube,Albizia and Juncus.The results show that there are 86 spectrum peaks in chromatogram.So far,46 components have been already identified,mostly of valepotrates iridoid,flavonoid glycosides and lignans glycosides.From the research,chemical information of non-volatile components in CAC have clearly been found,by the way,it has laid a solid foundation for analyzing the non-volatile chemical components in serum.2.Study on Pharmacochemistry of CAC in serumA method that preparation of rats serum containing CAC was established:giving rats with 6g/kg CAC twice a day in three consecutive days,drawing out blood from abdominal aorta of rats after the last lavage,drawing out 2ml supernatant after 12 hours,,and mixed with 6ml methanol,after centrifugation and dry with nitrogen,the 1.5ml remains also mixed with methanol,drawing out supernatant after vortex and centrifugation.From the results,41 components are found in serum,and 17 components have been identified,including of 14 prototype components and 3 metabolites.Most of identified components have been verified the anxiolytic effect in clinical or animal experiments.3.Performed a system pre-clinic pharmacokinetic study of(-)-Syringaresnol-4-O-?-Dapiofuranosyl-(1?2)-?-D-glucopyranoside in miceA sensitive and specific UPLC-TQ-MS/MS method was developed and validated for the quantification of SAG and SYR in biological samples of mice.A simple protein precipitation method with methyl alcohol is used to treat the biological samples,then the samples were saperated by using acetonitrile-water mobile phase.Electro spray ionization(ESI)source was applied and operated in negative ion modle;MRM 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 SAG and SYR in mice.Single oral doses of 15,30,and 60 mg/kg of SAG were given to mice respectively.UPLC-TQ-MS method was used for the quantification of SAG and SYR in mice plasma samples in different time.Plasma concentration-time data were analyzed by non-compartmental model with WinNonlin 4.1.The results showed that after single oral dose of SAG,the pharmacokinetics of SAG was fitted to rapid absorption,rapid distribution and slow elimination and was eliminated as first-order kinetics.The t1/2 value were 46.91,40.91,49.27min respectively;The Cmax value were 14.817,34.041,60.294ng/mL respectively;The AUC value were 1069.485,2276.715,3697.1925min × ng/mL respectively.A nonliner relationship was showed between the AUC and dose.The pharmacokinetics of SYR was eliminated as first-order kinetics,too.The t1/2 value were 46.91,45.91,50.27min respectively.Single oral doses of 60 mg/kg of SAG were given to mice.LC-MS method was used for the quantification of SAG 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,were all lower than the plasma drug concentration,suggesting that the low rate affinity of SAG with tissue.