Study On Separation Of Alkaloid And Spectrum-Effect Correlation Of Depression-Relieving And Spirit-Quieting Based On Compatibility From SZS

Objective: To optimizate the method for the determination of total alkaloids in SZS and purify it by high-speed countercurrent chromatography.To investigate the influences of different proportions of alkaloids,saponins and flavonoids on the behavior monoamine neurotransmitters in brain and hypothalamic-pituitary-adrenal(HPA)axis hormone levels in bold of in experimental depression.To investigate the best ratio of the role of Semen atagnation.To establish the fingerprints of different prorortions of SZS and the spectral-efficiency related mathematical models.Methods: 1.The puridied chemicals of total alkaloids was isolated with High-speed countercurrent chromatography.2.The HPLC fingerprint chromatography of total alkaloids and all diffent various proportion of three sets in SZS was established with HPLC-DAD method and the content of magnofiorine in total alkaloids was determined at the same time.3.Measure the stagnation and soothing effects on mice of different compatibility by establishing the CUMS mouse model;The contents of Glu,GABA,BDNF in the brain of mice and the CORT,ACTH,CRH in blood of mice was measured with Molecular Devices.4.According to the principle of correlation analysis,using SPSS to calculate the Spearman correlation analysis,gray correlation analysis and multiple regression analysis.Results: 1.Depending on the chemical properites of the alkaloid Alkaloids in SZS,selected butanol-ethyl acetate-water(3:2:5)as solvent system and isolated theree compounds,identified one of them was magnolia.At the same time,opidmized the fingerprint of alkaloid faracion in SZS,selectd magnolia as the reference peak,11 batches of samples were comapred and calibrated 18 common peaks whose number of peaks and resolution were incerased.2.Semen alkaloids and flavonoids can significantly improve the compatibility CUMS mice depressive symptoms of insomnia.Neuroendocrine neurotransmitter in mouse blood tests showed that the compatibility of alkaloids and flavonoids can reduce the contents of CRH,ACTH,CORT,suggesting that the antidepressant effects associated with decreasing brain nervous system excitatory mechanisms.The mouse brain neurotransmitters measurement results showed that alkaloids-flavonoids(1:14)compatibility can reverse the phenomenon of Glu increased and GABA decreased,suggesting that this group can improve the compatibility of depression with insomnia mice symptoms,and play a sedative effect;At the same time capable of raising CUMS mouse brain BDNF content.3.Based on fingerprints and efficacy indicators Spearman correlation analysis,gray correlation analysis and multiple regression analysis identified a comprehensive analysis,the peaks of compatibility Semen related with efficacy indicators retention time are alkaloid parts of 13.30 min,14.43 min,17.63 min and flavonoids parts of 20.69 min,23.91 min,26.20 min,37.93 min.And confirmed17.63 min to Magnolia base,26.20 min as spinosin,37.93min6’’’-Feruloylspinosin.Conclusion: In this study,the method of isolating alkaloid chemical by high-speed countercurrent chromatography was established with high recovery rate,easy operation and large separation rate.While the fingerprints conditions were optimized to improve the degree of separation between the various peaks,to establish a more perfect quality control standard of alkaloid in SZS.Pharmacodynamic Experimental results show that,Semen alkaloids and flavonoids(1:14)compatibility can significantly improve symptoms of depression with insomnia CUMS mice,which Atagnation mechanism may suppress the HPA axis,the balance of Glu and GABA levels increase brain-derived neurotrophic factor and other related mechanisms.Spectral efficiency by correlation analysis identified a total of seven major components associated with the pharmacodynamics,and has confirmed that ingredients are Magnolia,spinosin and 6’’’-Feruloylspinosin.