Analysis And Evaluation Of Active Ingredients In Zhejiang Planted Tetrastigma Hemsleyanum And The Study Of Its Antioxidant Activity

Objective The contents and their variances of polysaccharides and flavones(including total flavonoids and quercetin)in Zhejiang planted Tetrastigma hemsleyanum,and the antioxidant activity of alcohol extraction of the planted T.hemsleyanum and the correlation between flavones content and antioxidant activity were found out to provide scientific basis for substituting Zhejiang planted T.hemsleyanum for wild T.hemsleyanum,and for selecting the drying method,controlling the quality and researching and developing relevant medicine or health food of Zhejiang planted T.hemsleyanum.Methods(1)The project of L9(34)orthogonal test was designed to optimize the extraction process of polysaccharides and total flavonoids in artificial planted T.hemsleyanum,with the yields of polysaccharides and total flavonoids as evaluation indicators.Ultraviolet spectrophotometry method for the determination of polysaccharides and total flavonoids,and HPLC method for the determination of quercetin were established respectively.Polysaccharides,total flavonoids and quercetin contents of wild,artificial planted T.hemsleyanum were determined through the established methods.Besides,Polysaccharides,total flavonoids and quercetin contents of T.hemsleyanum collected from different localities,dried by different methods,and T.hemsleyanum at different growth stages were determined.(2)T.hemsleyanum was extracted by 70%ethanol,and separated by D-101 macroporous resin.Five different elution TDG-W、TDG-30%、TDG-50%、TDG-70%、TDG-90%were prepared respectively by washing out with different concentration of ethanol.Antioxidant activity of different elution and the correlation between flavones content and antioxidant activity were studied.Results(1)Polysaccharides,total flavonoids and quercetin contents of Zhejiang planted T.hemsleyanum were 20.69g/100g,I7.58mg/g,57.19ug/g,respectively,which was significant higher than that in wild T.hemsleyanum(15.28%,11.57 mg/g and 42.46ug/g,respectively).(2)Polysaccharides,total flavonoids and quercetin contents of Jiangxi planted T.hemsleyanum were 19.33%,12.98mg/g,43.07ug/g,respectively.Guangxi planted T.hemsleyanum were 15.1%,2.37mg/g,9.41ug/g,respectively.Fujian planted T.hemsleyanum were 13.93%,12.23mg/g,37.34ug/g,respectively.Zhejiang planted T.hemsleyanum were 20.69%,17.58mg/g,57.19ug/g,respectively.Therefore,Polysaccharides,total flavonoids and quercetin contents of T.hemsleyanum from different localities were significantly different,and the contents of Zhejiang planted T.hemsleyanum was the highest.(3)The losses of polysaccharides,total flavonoids and quercetin of Zhejiang planted T.hemsleyanum caused by traditional hot air drying were 17.18%,20.03%and 22.13%respectively,caused by vacuum drying were 13.41%,23.79%,9.40%,respectively.However,the losses caused by freeze drying were tiny for polysaccharides,total flavonoids and quercetin contents of freeze dried T.hemsleyanum were close to that of fresh T.hemsleyanum.(4)Polysaccharides and total flavonoids of Zhejiang planted T.hemsleyanum at different growth stages were significantly different.Polysaccharide contents achieved the highest in December 2014,and total flavones achieved the highest in February 2015.(5)Total flavonoids contents of different elution TDG-W,TDG-30%,TDG-50%,TDG-70%,TDG-90%were 2.69%,33.25%,61.12%,40.54%,4.12%,respectively.The antioxidant activity of elution TDG-50%was the highest among all elution,its DPPH free radical scavenging ability,reducing capacity and total antioxidant capacity of the elution TDG-50%were 87.60%(75μg/mL),1.627(400μg/mL),1.812(1000μg/mL),respectively,higher or close to that of positive control BHT(77.68%(75μg/mL),1.893(400μg/mL),0.436(1 000μg/mL),respectively).Conclusion(1)the main active ingredients of Zhejiang planted T.hemsleyanum were higher than that of wild T.hemsleyanum,so Zhejiang planted T.hemsleyanum can be used as a substitute for wild T.hemsleyanum in clinical practice.(2)Taking into account the quality control of main active ingredients like polysaccharides and total flavonoids of T.hemsleyanum,it’s the highest content of the main active components in the period of growth for 3 years or more,for the period from December to February each year to harvest the best quality.(3)Freeze drying can maximumly preserve the active ingredients of fresh T.hemsleyanum,so it is the best drying method for T.hemsleyanum.(4)The total flavonoids contents of different alcohol elution were different,and they all had strong antioxidant capacity.Moreover,the DPPH free radical scavenging capacity,reducing capacity and total antioxidant capacity of alcohol elution increased with the increase of total flavonoid concentration.In conclusion,this study provides scientific basis for the preservation of active ingredients and the selection of drying method of T.hemsleyanum,and quality control,research and development of relevant medicine or health food of artificial planted T.hemsleyanum.