The Extraction Process Optimization, Ingredient Preparation And Antioxidant Activity Evaluation Of Mongolian Medicine "Di Sauer Gourd"

Objective:The study was aimed to optimize the water extraction and ethanol extraction for total flavonoids from Cynanchum Thesioides(Freyn.)K.Schum by orthogonal array design;to study the external in vitro antioxidant activity of two different extracts,respectively;and to systematically purify and identify the chemical constituents from the Cynanchum Thesioides(Freyn.)K.Schum and obtain the compound.Experiment methods:By single factor experiment and orthogonal,with total flavonoids as the indicator,the effects of ethanol concentration,extraction time,liquid-to-material ratio and soaking time on the extraction effect were investigated,and the extraction process was optimized.By testing the ability to scavenge DPPH free radical scavenging method and the protective effects of different extracts on DNA oxidative damage induced by OH and hydrogen peroxide,in vitro antioxidant activity of water extract and ethanol extract were determined.Using both of the ethanol and water as solvents,Cynanchum Thesioides(Freyn.)K.Schum was extracted and concentrated by heating and refluxing.Put the obtained extracts solve in the water and extract with petroleum,ether n-butanol and dichlormethane successively.On the basis of HPLC chromatograms,the fraction eluted from the parts of ether n-butanol and dichloromethane were separated on macoporous resin HPLC to obtain column,silic a gel column,polyamide column,silica gel preparation plate and monomer compounds.The molecular formulas and structures were identified by MS,1H-NMR,13C-NMR,2D-NMR and the others spectroscopic methods.Experiment results:In the water extraction,the best conditions of total flavonoids were 120 min of extraction of time,30:1 of liquid-to-material ratio,2 hours soaking time.The optimal ethanol extraction technology of total flavonoids was 60%of ethanol,30:1 of liquid-to-material ratio,2 hours of soaking and 1.5 hours of refluxing.The IC50s of water extracts and ethanol extracts were 0.3449 mg/ml and 0.2472 mg/ml respectively.The increase of fluorescence intensity was used to determine the extracts-repaired damaged DNA double strand structures which were previously damaged by OH and H2O2.The antioxidant activity of the alcohol extract is stronger than that of the water extract,and the antioxidant capacity is continuously enhanced as the concentration increases.Eleven compounds were isolated and six compounds were identified,including:thesioideoside,quercetin-3-O-galactoside,dihydroquercetin,quercetin,tamarixetin 3-O-(?-D-galactopyranoside)and adenosine.Experiment conclusion:The laboratory level technology of water extraction and alcohol extraction of the Cynanchum Thesioides(Freyn.)K.Schum was successfully established.Both of the water extract and alcohol extract have the active ability of eliminating free radicals in vitro,while the antioxidative effect of alcohol extract was better.The flavonoids(dihydroquercetin and quercetin),flavonoid glycosides(quercetin-3-O-galactoside,tamarixetin and 3-O-(?-D-galactopyranoside)and adenosine was isolated successfully from Cynanchum Thesioides(Freyn.)K.Schum.It is the first time for dihydroquercetin,tamarixetin and adosine discovered from Cynanchum Thesioides(Freyn.)K.Schum.