Material Basis Of The Husks Of Xanthoceras Sorbifolia Bunge In The Treatment Of AD Disease And Pharmacokinetic Study

In this study,the husks of Xanthoceras sorbifolia Bunge was selected as the objective of the research,and the following aspects were achieved and deeply investigated:1.UPLC-Q/TOF MS was applied to identify chemical compositions and the constituents in husks of WG.29 compounds were identified in WG,and 15 constituents of them are triterpenoid saponins,4 constituents are flavonoids,2 constituents are coumarin,4 constituents are phenolic acids and other 4 other constituents.Then,make a summary of the cracking rules of each compound.Among the 29 constituents,15 of them were identified by comparation with the reference standard through the retention time and relative molecular mass.2.An HPLC-MS method was applied to simultaneously determine 7 compounds in 7 different parts of WG.The 7 compounds were separated by Phenomenex C18 Column(250 mm × 4.6 mm,5μm)column.The mobile phase was 0.05%formic acid water-acetonitrile.And they were detected in SIM mode with ESI+.The method was examined and found to be satisfied with linearity,precision,accuracy,recovery and stability for all analytes.This study may be helpful for the quality control of WG.3.To investigate pharmacodynamics in rats after oral administration of WG.The AD model was induced by intraperitoneal injection of D-Gal and bilateral hippocampal injection of Aβ25-35.2 weeks after surgery,the behavioral test and brain tissue pathological biopsy were carried out.The behavioral test showed that WG group significantly(P<0.05)improve percent alternation in Y maze task and decrease escape latency in the directional swimming test in AD rats.And it could also prolong the swimming distance and time in the target quadrant and around the platform 10 cm,and increase the number of across the platform in rats with D-Gal and Aβ25-35 in probe test.Brain tissue pathological biopsy showed that the CA1 hippocampal cells of rats in WG group were gradually returned to normal.All experimental results show that WG has prevention and treatment function on AD.4.The rapid and sensitive UFLC-MS/MS methods for the quantification of DA,NE,Ach,5-HT,Glu,GABA,Trp and Asp in rat brain without derivatization or complex sample pre-treatments were developed.After addition of the internal standard,isoproterenol,the samples were extracted by protein precipitation.The intra-day and inter-day precision and accuracy of the analytes were well within acceptance criteria(±15%).The mean extraction recoveries of analytes and internal standard were all more than 60.00%.The methods were examined and found to be satisfied with linearity,precision,accuracy,recovery,matrix effect and stability.The methods were succefully applied in the determination of related substances of rats in blank control,model,WG and Hup-A group.The results demonstrated that compared with the blank group,the concentrations of NE,Ach and Glu in model rat brain were significantly decreased(p<0.001),the brain Trp level was significantly increased;compared with the model groups,the brain Ach was increased and NE,Glu,Trp were descreased(p<0.001).DA,NE,Ach,5-HT,Glu,GABA,Trp and Asp concentration in WG group were recoveried to control group and were similar with Hup-A group.So WG may adjust the level of the neurotransmitter and have a role in prevention and treatment of AD.5.A fast,sensitive and reliable HPLC-MS method has been developed and validated for simultaneous quantitation of compound 1,2,3,5,7 in rat plasma after oral administration of the 70%ethanolic extract of the husks of yellowhorn.The plasma samples were extracted by liquid-liquid extraction using ethyl acetate-isopropanol(1:1,v/v).Good chromatographic separation was achieved using gradient elution with the mobile phase consisting of 0.05%formic acid in water and acetontrile.The tandem mass spectrometric detection was performed in SIM mode on HPLC-MS system with turbo ion spray source in a positive ionization mode.Both intra-day and inter-day precision and accuracy of analytes were well within the acceptance criteria(15%).The mean absolute extraction recoveries of analytes and IS from rat plasma were all more than 60.0%.The validated method has been successfully applied to pharmacokinetic profiles of analytes.The results indicated that compound 1-3,5,7 absorbed very quickly,tmax were less than 1.5h.The mean plasma concentration-time profiles of WG were drafted and showed the douple peak phenomenon.