The Viola Appraisal From Web With ITS And Polysaccharide Optimization And Medical Research Of Viola Inconspicua Blume

Viola Inconspicua Blume is a Violaceae Viola,a perennial herb,which mainly glows in the roadside trees,grass slopes environment.As is reported in the literature,Viola inconspicua Blume is also the largest amount of resources of the genus in the Southern China.In the folk,Viola is ususlly used by whole plant,they have Qing re jie du,swelling,pain,the treatment of hepatitis and throat inflammation effect,a number of research work at China and abroad show that the genus containing polysaccharides and other medicinal ingredients.reactive oxygen and nitrogen plays an important role forthe organism,which is in the dynamic balance of the human body.oxidative stress too much on the cell structure and function of biological macromolecules will cause significant damage,which is directly or indirectly caused by aging and some diseases.In recent years,on the antioxidant system in vitro,polysaccharides or glycoconjugates which derived from natural drugs is a free radical scavenger,effective reducing agent and iron chelator,and rational utilization of these natural,non-toxic,no side effects of antioxidants,has considerable application value for the prevention of the disease and effective antioxidant function in vitro and in vivo.This is the first time in detail on the inconspicua Blume polysaccharide extraction and determination of content,on the basis of the experiment,using response surface and Box-Benhnken method combined with experiment.Purification and characterization of the optimal extraction conditions of polysaccharides and separation,and in vitro antioxidant activity and cytotoxicity evaluation.The main results are as follows:1.The violet traditional medicinal material of herba and tianshan violet has the obvious difference of molecular systematics from Viola inconspicua.2.By ultrasonic water extraction and ethanol precipitation of polysaccharide,crude polysaccharide content is 3.55%.The response surface test showed that the optimum extraction technology of polysaccharide: the extraction solvent is distilled water,the extraction temperature is 100 ℃,extraction time 4 h,extraction rate is 2 times,theory of polysaccharides optimal value is 5.51%.3.Using Savag reagent to remove protein,after using gradient dialysis bag to dialysis,next waiting for DEAE-Sphere ion exchange chromatography resin water balance,by eluting with distilled water and 0.1~1M Nacl solution,phenol-sulfuric acid method of each polysaccharide content in liquid collected was detected,The polysaccharide was separated by gel chromatography and eighteen angle laser light scattering system,and the molecular weight,molecular weight and rotation radius were determined,it has been determined that the molecular of polysaccharide content mostly be 2.086×10~6 g/mol.4.The antioxidant activity in vitro of the polysaccharide experimental results show that in the concentration range of 1~5 mg/ml,2,2ˊ-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)radical(ABTS+·),1,1-diphenyl-2-picrylhydrazyl(DPPH)has the function of eliminating and reducing power of iron oxidation(FRAP)reduction.And with the increase of the concentration of polysaccharide,its removal and reduction has increased,the inconspicua Blume polysaccharide concentration was 5 mg/ml,ABTS+ clearance rate was 48.3%,DPPH free radical scavenging rate is 58%,reducing power for FRAP is(1.75 ± 0.04)mol Trolox/mg.