Study On Pharmacokinetics And Tissue Distribution Of Multiple Active Ingredients In Xiaoyao Powder

Xiaoyao Powder?XYP?is a traditional Chinese medicine,derived from the Song Dynasty Taiping Huimin Heji Jufang.It is a classical Chinese herbal formula for liver stagnation,invigorating spleen and stomach,nourishing blood and regulating menstruation.The pharmacology,pharmacodynamics and chemical constituents of single herbs have been deeply studied,but the pharmacokinetics and tissue distribution of XYP have not been reported.To provide scientific basis for clinical rational application of XYP,the pharmacokinetics and tissue distribution of the main active ingredients of XYP were studied.The 14 main active ingredients?albiflorin,paeoniflorin,ferulic acid,senkyunolide I,quercetin,isoliquiritigenin,atractylenolide III,ligustilide,atractylenolide II,liquiritin,liquiritigenin,saikosaponin c,glycyrrhizic acid,and saikosaponin a?in XYP were selected as the indicators of multicomponent pharmacokinetics.In this study,uplc-ms method was used to detect the index components of female rats after oral XYP,so as to obtain the pharmacokinetic parameters and tissue distribution characteristics of 14 main active components.In this study,the compounds were separated using an ACQUITY UPLC^TM BEH C18 column?1.7?m,50 mm×2.1 mm?with a mobile phase consisting of acetonitrile and 0.1%formic acid in water at a flow rate of 0.3 mL/min.Detection was performed on a triple-quadrupole tandem mass spectrometer using multiple reaction monitoring and an electrospray ionization source in both positive and negative ionization modes.The results showed that the accuracy,precision and stability of the UPLC-MS/MS method were up to the standard and could be used to calculate the pharmacokinetic parameters and study the tissue distribution of 14 target components.All calibration curves exhibited good linearity?r²>0.9974?over the measured ranges.The intra-and inter-day precisions were within 12%,and the accuracy ranged from 89.93%to106.64%.Extraction recovery is over 82.33%,matrix effect is between 85.17 and102.69%.This method is suitable for the basic requirements of quantitative analysis of components in vivo.1.Pharmacokinetic experimentsAfter oral administration of XYP extract?4 g/kg?for 0?0.083?0.17?0.33?0.5?0.75?1?2?4?8?12 and 24 h?n=6?,1 mL of blood was collected from orbital venous plexus respectively.After treatment,the contents of 14 target substances were analyzed by UPLC-MS/MS,and the main pharmacokinetic parameters were calculated by Winnolin 3.2 software.The results showed that ferulic acid,quercetin and liquiritigenin entered blood rapidly and absorbed quickly in rats(T_max 0.10±0.03,0.21±0.10,0.19±0.0).Atractylenolide II and atractylenolide III absorbed blood more slowly than other target analytes(T_maxax 0.64±0.29,0.67±0.26).The concentration-time curves of flavonoid aglycones?liquiritigenin,isoliquiritigenin and liquiritin?and triterpene glycosides?glycyrrhizic acid,saikosaponin a andsaikosaponin c?all showed double peaks.The concentration of atractylenolide II and atractylenolide III in XYP extract was similar,but the absorption of atractylolide III in rats was much higher than that of atractylolide II(AUC_0-t and C_max were about ten times different).2.Tissue Distribution ExperimentAfter oral administration of XYP extract?4 g/kg?,rats were decapitated at 0.25?0.5?1?2?4 h?n=6?.Heart,liver,spleen,lung,kidney,brain,stomach,uterus,ovary and small intestine were taken immediately.After sample pretreatment,tissue samples were analyzed by UPLC-MS/MS.The results showed that atractylenolide III,ligustilide and senkyunolide I could be detected in brain tissue;albiflorin,paeoniflorin,quercetin,ligustilide and atractylenolide II,were higher in liver and spleen;albiflorin,paeoniflorin,senkyunolide I,quercetin,atractylenolide III,ligustilide,liquiritin,liquiritigenin and glycyrrhizic acid were higher in ovary and uterus;ferulic acid,quercetin,isoliquiritigenin,atractylenolide III,atractylenolide II and saikosaponin a were higher in stomach and intestine.The multicomponent pharmacokinetic study of XYP showed that the absorption rates of 14 analytes varied greatly in vivo,which was related to the structure category and relative configuration of the compounds.The tissue distribution experiment of XYP confirmed that 14 analytes were mainly distributed in ovary,uterus and liver.This result is consistent with the characteristics of XYP in treating liver depression syndrome and perimenopausal syndrome.This study reveals for the first time the main absorption and distribution of XYP after oral administration of blood,laying a material foundation for the theoretical study of XYP and the secondary development of classical prescriptions.