Antidepressant Effect And Separation And Enrichment Of Chemical Components From The Extract Of Ormosia Henryi Prain Leaf

Ormosia henryi Prain is an evergreen tree of the family Leguminosae and genus Ormosia.Folk research found that its leaf is a new biological resource with potential antidepressant activity.However,it lacks modern scientific research and its chemical composition has not been reported.In this study,Ormosia henryi Prain leaf extract(OHPLE)was used as a raw material to study its antidepressant effect using a mice model,and its safety was initially determined by an acute toxicity test in mice.Then,eight flavonoids were separated and identified from the extract by using modern separation and purification technology.On this basis,the ultra-performance liquid chromatography/electrospray ionization quadrupole timeof-flight mass spectrometry(UPLC-ESI-QTOF-MS/MS)technology was further used to analyze other chemical components of the extract,and a total of 46 flavonoids were identified or preliminarily speculated.Finally,the extraction and purification processes of total flavonoids from Ormosia henryi Prain leafs(OHPL)were optimized to provide reference for the development and utilization of OHPL.The main findings are as follows:(1)The antidepressant effect of OHPLE was studied in mice model of chronic unpredictable mild stress(CUMS).The results showed that high dose(150 mg/Kg)of OHPLE could significantly increase the index of sucrose preference and shorten the ingestion latency in mice.Middle and high dose(100 mg/Kg and 150 mg/Kg)of OHPLE significantly increased the activity time and reduced the rest time of mice in tail suspension test,and significantly increased the content of brain derived neurotrophic factor(BDNF)in the hippocampal tissues of mice.The results showed that OHPLE had significant antidepressant effect.(2)The acute toxicity test of OHPLE showed that there was no obvious abnormal autonomic activity in mice within 0～4 hours after oral administration of OHPLE.After 14 days of observation,there was no significant difference in body weight compared with the control group,no obvious abnormalities were observed on the surface and section of each organ,and there was no death.No obvious toxic reaction was observed at 10 g/kg(250 mg/mL).(3)Eight flavonoids were isolated from OHPLE by high-speed counter-current chromatography and preparative high-performance liquid chromatography(HSCCC-PreHPLC).The structure was identified by mass spectrometry and nuclear magnetic technology.They were isoorientin-2′′-O-rhamnoside,isoorientin,orientin,vitexin-2′′-O-rhamnoside,isovitexin-2′′-O-rhamnoside,isovitexin,diosmin,linarin,which were the first reports of this plant.(4)Using UPLC-ESI-QTOF-MS/MS technology and combined with the analysis of the mass spectrometry cracking rules of the separated compounds,further analysis of multiple chemical components in OHPLE.A total of 46 flavonoids were identified or preliminarily speculated,including 9 flavone C-glycosides,11 flavones,5 flavone O-glycosides,5isoflavones,5 isoflavone O-glycosides,8 prenylflavones and polymethoxyflavones.(5)The extraction process of total flavonoids from OHPL was optimized by reflux extraction method.Based on the single-factor test,the optimal extraction process parameters finally determined by the orthogonal test were: ethanol volume fraction 60%,material-liquid ratio 1:40(g/mL),extraction temperature 80 ℃,and extraction time 1.5 h.Under this process,the yield of total flavonoids from OHPL was 2.83%,and the total flavonoid content in the dry powder of the extract is 15.95%.(6)The macroporous adsorption resin method was used to optimize the total flavonoid enrichment in OHPL.Through the investigation of the static adsorption and desorption capabilities of ten macroporous adsorption resins(HPD-100,HPD-300,HPD-400,HPD-600,HPD-826,DM130,AB-8,D101,NKA-9,X-5),the AB-8 macroporous adsorption resin was selected for process optimization.The optimal purification process parameters were determined as follows: the concentration of flavonoids in the sample solution was 1.71 mg/mL,the pH value of the sample solution was 3,the flow rate of the loading was 4 BV/h,the volume of the loading was 5 BV,the volume fraction of eluent ethanol was 80%,the flow rate of elution was 3 BV/h,and the volume of elution was 4 BV.Under this process condition,the total flavonoids content in the purified drying products was 29.51%.