Studies On Absorption And Pharmacokinetics Of Bioactivity Compounds Of Zishen Pill

Zishen pill was composed of Anemarrhenae Rhizoma(zhimu), PhellodendriChinense Cortex(huangbo) or Phellodendri Amurensis Cortex(guanhuangbo) andCinnamom Cortex(rougui) with a ratio of10:10:1in weight. It showed good activityboth in clinical and in animal experiments. Our previous study showed that in prostate,the receptor organ of BPH, xanthone glycosides, timosaponins and alkaloids were themajor constituents occurred. While the substantial foundation and pharmacologicalmechanism of ZSP was still unclear, which hampered the systematically recognitionof ZSP.Our study was mainly focused on systemically analysising the chemicalcompounds of ZSP. Besides, the major components and metabolites in rat biologicalspecimens after oral administration of ZSP were quantitative and quantified. Thespecific contents are as followed:Firstly, identification of the chemical components of ZSP and the bioactivecomponents and metabolites in rat biological specimens after oral administrationof ZSP.An ultra high performance liquid chromatography coupled with electrosprayionization quadrupole time-of flight mass spectrometry (UHPLC-ESI-Q-TOF/MS)method has been applied to comprehensively screen the chemical compounds of ZSPand bioactivity compounds of and metabolites in rat biological specimens after oraladministration of ZSP. Through the analysis, a total of101compounds have beenidentified or tentatively characterized from the Zishen pill, including alkaloids,xanthones and timosaponins.18of them were ambiguously identified through a directcomparison of the retention time and MSnfragmentation patterns with the authenticones.17absorbed chemical constituents of Zishen pill and10metabolites wereidentified in plasma,13chemical constituents and10metabolites in prostate,33chemical constituents and22metabolites in urine. Among the55ions detected inurine,33of them were from Phellodendri Amurensis Cortex and22of them weredetected from Anemarrhenae Rhizoma. Among those metabolites,18of them werephase II metabolites and4were phase I metabolites, methylation, glycosylation,glucuronidation and sulfation are the major metabolic processes of xanthones. Andoxidative, demethylenation and glucuronidation are the major metabolic pathways ofalkaloid. Besides, a new UHPLC-Q-TOF/MS coupled with mass profiler method hasbeen successfully developed. Secondly, Separation and purification of phase I metabolites of berberine bymass-based preparative LC method.Our previous studies showed that alkaloids were the major activity constituentsof ZSP, but the pharmacokinetic of chemical constituents of ZSP can not elucidate themechanism of ZSP. Thus, the effect of metabolites should be taken into consideration.Metabolites of berberine, including berberrubine and demethyleneberberine, weresynthesized and successfully prepared via preparative LC employing mass-basedfraction collection.Thirdly, Pharmacokinetic study of the bioactive components and theirmetabolites in rat plasma after oral administration of ZSPA UHPLC-MS/MS method has been developed to simultaneous quantitativedetermination of7bioactive components (berberine, palmatine, phellodendrine,magnoflorine, timospaonin EI, timosaponin B-II and timosaponin B) and3metabolites (demethyleneberberine, thalifendine and berberrubine) in rat plasma afteroral administration of ZSP. During analyzing, magnoflorine showed highconcentration in plasma, which drop a hint that aporphine alkaloid might play animportant role in ZSP.