Study On The Extraction And Identification Of Antioxidant Active Components From Atractylodes Macrocephala

Atractylodes macrocephala has great development value in the pharmaceutical industry and health care industry.The extraction and identification of the antioxidant active ingredients of Atractylodes macrocephala were studied and a theoretical reference for the development and application of Atractylodes macrocephala was provided.This article is divided into three parts.(1)Optimization of extraction conditions for antioxidant active ingredients of Atractylodes macrocephala Koidz.Taking the yield and antioxidant activity of extracts of Atractylodes macrocephala Koidz as the indexes,the methanol volume fraction,solid-liquid ratio,extraction time,and extraction temperature were selected as independent variables,single factor experiment and response surface methods were used to optimize the extraction process of Atractylodes macrocephala Koidz.The results showed that: The volume fraction of methanol is 87%,the ratio of liquid to material is 35(ml/g),the extraction temperature is 62°C,and extraction time is 1.5h.Under the above conditions,the yield of Atractylodes macrocephala is 42.23%,which is basically consistent with the expected value 42.25% of the model.The antioxidant activitiy of extract of Atractylodes macrocephala under the optimal extraction conditions was tested.The Ic50 value of scavenging DPPH free radicals was 0.238 g/l,and the clearance rate was better.(2)Determination of the antioxidant activity parts of Atractylodes macrocephala extractIn order to further study the antioxidant active ingredients of Atractylodes macrocephala,87% methanol extracts of Atractylodes macrocephala were successively extracted with petroleum ether,chloroform,ethyl acetate and n-butanol to obtain different solvent extracts.The abilities of eliminating ABTS radicals and DPPH radicals and total reducing capacity were the evaluation indicators,the antioxidant activities of the extracts were studied,and the relationship between the antioxidant activities and total polyphenols,total flavonoids,and total polysaccharide contents were explored.The results showed that different solvent extracts of Atractylodes macrocephala Koidz had antioxidative activities.The ethyl acetate phase had the best antioxidant activity,the Ic50 values of ABTS radical and DPPH radicals were 0.021±0.003mg/ml and 0.031±0.005,respectively.The contents of total polyphenols and total flavonoids in the ethyl acetate phase were the highest,which were 37.5±0.51% and 30.5±0.92%,respectively.There were correlations between the total reduction,Ic50 value of scavenging DPPH,ABTS free radicals and the total polyphenol content(r=-0.901,r=-0.870,r=0.969).The total reducing power,Ic50 values of scavenging DPPH free radicals and ABTS were significantly correlated with total flavonoids.(r=-0.912,r=-0.976,r=0.913).Flavonoids and polyphenols in the extracts of Atractylodes macrocephala may be the main antioxidants.(3)Identification and content determination of antioxidant active ingredientsIn order to trace the specific antioxidative substances in Atractylodes macrocephala Koidz,eliminating DPPH free radicals as an indicator,ethyl acetate of the Atractylodes macrocephala was further separated by column chromatography to obtain the best active sample.The sample was analyzed by high performance liquid chromatography and mass spectrometry,The contents and antioxidant activities of identified chemical constituents were determinated.The results showed that the C5 component has the best anti-oxidation activity with Ic50 value of 0.021(mg/ml).After comparing with standard products by mass spectrometry,high performance liquid chromatography,C5 contains six compounds,namely chlorogenic acid,caffeic acid,ferulic acid,isovitexin,isochlorogenic acid A,and isochlorogenic acid C,the contents of the six compounds in order: isochlorogenic acid A> isochlorogenic acid C> chlorogenic acid> ferulic acid> isocamptocrine glycoside> caffeic acid,which were 80.26(mg/g)and 77.79(mg /g),38.12(mg/g),3.31(mg/g),1.21(mg/g),1.14(mg/g)respectively six compounds.The order of antioxidant activity was: isochlorogenic acid C> isochlorogenic acid A> chlorogenic acid> ferulic acid> caffeic acid> isoimpurine glycoside.