The Study On Metabolism Of Nevadensin In Vitro And In Vivo And Pharmacokinetics Of Compound Lysionotus Pauciflorus In Rats Based On LC-MS/MS Technology

Objective:A strategy was firstly developed to identify the metabolites of nevadensin in vitro and in vivo by UHPLC-Q-TOF-MS/MS.A LC-MS/MS method was developed for quantitative determination of six active components from compound of Lysionotus pauciflorus Maxim.To develop a LC-MS/MS method for quantitative determination of five active components from compound of Lysionotus pauciflorus Maxim.in rat plasma and applicate to its pharmacokinetic study.A sensitive,efficient,and rapid LC-MS/MS method was developed to simultaneously determine two pairs of isomeric flavonoid glycosides in rats.Methods:An on-line data acquisition method a multiple mass defect filter combined with dynamic background subtraction was developed to trace all probable metabolites.Furthermore,some assistant tools,such as key fragment ions,were employed for compound hunting and identification.The chromatographic separation was carried on Poroshell 120 EC-C₁₈ 2.1 x100mm,2.7?m)with a SecurityGuard^?UHPLC C₁₈ pre-column?Poroshell?.Mass spectrometric detection was performed on a Triple TOF^TM 5600 system with Duo-Spray^TM ion sources with an electrospray ionization?ESI?source in the negative ion detection mode.Chromatographic separation was achieved on a Wonda Cract ODS-2 C₁₈ Column?150 mm x 4.6 mm,5?m?.The detection was performed in negative electrospray ionization mode and the precursor/product ion transitions of six components and internal standard sulfamethoxazole were quantified in multiple reaction monitoring using QTRAP-3200 MS/MS.The chromatographic separation was carried on a Wonda Cract ODS-2 C₁₈ column?150 mm x 4.6 mm,5?m?.The detection of four analytes and internal standard sulfamethoxazole was performed with multiple reaction monitoring in negative electrospray ionization mode.Results:A total of 23 metabolites were structurally characterized in vivo including 16 phase ? and 7 phase ? metabolites,and 12 metabolites were detected in vitro containing 10 phase ? and 2 phase ? metabolites.The results indicated that oxidation,hydrolysis,demethylation,methylation,sulfate conjugation and glucuronide conjugation were main metabolic pathways of nevadensin.According to the identification of the major components,nevadensin was the most abundant followed by rosemary acid.The result indicated that nevadensin and rosemary acid were the main active components.After oral administration of 10 mL/kg compound extract in rats,nevadensin and rosemary acid were the main absorption components in plasma.The pharmacokinetic study indicated that analytes reached their Cmax in about 15 minutes.Conclusions:It was the first time the method of incubation in liver microsomes was applied to the study of nevadensin.Overall,it was also the first study of the metabolic mechanism of nevadensin in vitro and in vivo.This study maybe can provide reference and valuable evidence for further investigation of the metabolic mechanism of nevadensin.A selective and sensitive LC-ESI-MS/MS method was established and validated in rat plasma for the first time.The pharmacokinetics of the compound of Lysionotus pauciflorus Maxim.in rat plasma was first reported.The pharmacokinetic parameters and plasma concentration-time profiles would prove valuable in pre-clinical and clinical investigations on the disposition of compound medicine.This method was successfully applied to simultaneous determination of the concentrations of four compounds in plasma after oral administration of 10 mL/kg Commelina communis Linn extract in rats.The method was validated to meet the quantitative analysis criteria required for the sample.It was the first report on pharmacokinetic studies of Commelina communis Linn.The information accomplished from this research may be valuable for the pre-clinical and clinical application of Commelina communis Linn.