Study On The Extraction And Antioxidant Activity Of Polysaccharides And Flavonoids Of Chrysanthemum

Chrysanthemum Morifolium Ramat.,which belongs to the head flower of herbal plant in Chrysanthemum family and is usually used as Chinese traditional perennial herbal medicine and drinking tea has multiple pharmacological actions,namely antioxidant,antitumor,preventing aging and great resource that has great developmental potential advantage.The major active components of Chrysanthemum Morifolium Ramat.are polysaccharides and flavonoids including acacetin,luteolin,quercetin,hesperetin,myricetin,apigenin.However,a few studies on the antioxidant activity evaluation of chrysanthemum polysaccharides were reported,lacking systemical examination on the extracting methods for the flavonoids of chrysanthemum.There were also few studies on the correlation between antioxidant capacity of chrysanthemum flavonoids extract and the contents.So it has great potentiality to be exploited.This article will conduct an in-depth study on the extraction of effective active components of chrysanthemum flower and their antioxidant effect,preliminarily investigating chromatographic pharmacodynamic relationship of chrysanthemum.The main contents are as follows:1.Response Surface Analysis for Optimizing Extraction of Crude PolysaccharidesWater extraction was found to be the suitable process for the extracting of the crude polysaccharides from chrysanthemum through the extraction percent yield being the standard index.One-factor experiment and response surface methodology were utilized with the consideration of actual operation and the optimal extracting conditions for the polysaccharide compounds were as follows:85? of extracting temperature,3h of extracting time,80%of ethanol terminal concentration,25:1(ml/g)of liquid to solid ratio under the fixed conditions:grinding particle size 60?m,water extraction,extracting for three times and pH value of 7.0.2.The Measurement of Antioxidant Activity Index of Crude Polysaccharides Extracted by Optimization ProcessThe chrysanthemum crude polysaccharides extracted by optimal water extraction process based on response surface methodology were measured by the antioxidant effects.Results showed that they were able to scavenge different free radicals including DPPH·,O2·-,-OH,ABTS·+and the antioxidant activity was proportional to the amount of polysaccharides content within the certain concentration range.3.Extraction Process for FlavonoidsBy using the percent yield of flavonoids extraction and the ability to clear DPPH-as the standard indexes,the six extraction processes were ordered from the highest percentage of flavonoids to the lowest as follows:methanol>95%of ethanol(original process)>70%of ethanol>boiling water>acetone>ethyl acetate;the following extraction processes were arranged from the biggest DPPH removal ability to the smallest:methanol>70%of ethanol>95%of ethanol(original process)>boiling water>acetone>ethyl acetate and the DPPH-clearing activity was proportional to the amount of flavonoids content within the certain concentration range.With the safety and cost-effectiveness of reagents for consideration,70%ethanol extraction process was regarded as the ideal method for chrysanthemum flavonoids.4.The Establishment of HPLC-UV Fingerprints and the Measurement of Antioxidant Activity Index of Flavonoids ExtractUnder the certain chromatographic conditions,using standard samples as the control,the fingerprints of flavonoids extract from thirty batches of Chrysanthemum Morifolium Ramat.dry flowers were detected,among which six common peaks were calibrated.And the six compounds were qualitatively identified as follows:quercetin,hesperetin,acacetin,myricetin,luteolin,apigenin.Meanwhile,the HPLC-UV method in good methodology validation was established for the content of the six kinds of flavonoids.According to the method,the amount of flavonoids from different batches of chrysanthemum was identified,laying a good foundation for the further research and development of chrysanthemum.The antioxidant effects of flavonoids extract of thirty batches chrysanthemum samples were determined by the method of DPPH·scavenging.Results showed that the flavonoids extract have strong antioxidant activity,and different batches have different DPPH· scavenging ability.5.Building the Correlation Model between HPLC-UV Fingerprints of Flavonoids and Antio-xidant ActivityThe method of grey related analysis was used to calculate the grey correlation degree between each chromatographic peak among flavonoids and the antioxidant activity.By the multiple stepwise regression analysis,five peaks which were statistically significant(P<0.05)to antioxidation were screened.Spectrum-efficiency relationship between the fingerprint of chrysanthemum flavonoids and the antioxidant activity could be analysed.