Investigation Into Purification,Bacteriostasis And Immunomodulatory Activity Of Anthocyanins From R.Setate×R.Rugosa

This study mainly takes the dried R.Setate×R.Rugosa as raw materials,focusing on the parameter extraction and purification process of anthocyanins from R.Setate×R.Rugosa,and further studied the antibacterial activity and immune activity of anthocyanins from R.Setate×R.Rugosa.It provides a scientific basis for the development and development of natural,efficient and non-toxic food and drugs.The contents of the study are as follows:(1)the content of anthocyanins in the extract as the index,based on six single factor experiments,using Box-Behnken response surface optimization method,get the best extraction process parameters for anthocyanins from R.Setate×R.Rugosa: extract pH=1.05,ethanol concentration 81.5%,extraction time 45 min,ratio of material to liquid 1:90(g/mL),extracting 2 times.The extraction rate of 4.27mg/g.(2)the static adsorption and analytical ability of D-101 macroporous resin on anthocyanins from R.Setate×R.Rugosa were discussed,It was found that the separation and purification of D-101 macroporous resin was excellent.On the basis of static test,the best conditions for dynamic purification are: the sample liquid pH=2,the adsorption velocity of1 m L/min,the sample liquid concentration 0.15mg/mL,the adsorption rate is 76.25%,eluent acidity 95% ethanol,30 mL dosage of eluent.The anthocyanins were all eluted under this condition.After purification,the purity of rosin was 4.1 times higher and the purity was up to35.62% after two purification,and its purification effect was remarkable.(3)the Bacteriostasis of anthocyanins from R.Setate×R.Rugosa was preliminarily explored by filter paper method.Research shows that: anthocyanins from R.Setate×R.Rugosa on Escherichia coli,Salmonella had no obvious inhibitory effect,but for staphylococcus aureus showed strong antibacterial ability,and its strength with the concentration increased,further showed the minimum inhibitory concentration of anthocyanins from R.Setate×R.Rugosa on Staphylococcus aureus(MIC): 1.0 mg/mL(4)Intraperitoneal injection of cyclophosphamide(CTX)to construct the immunosuppressive mice model,then intragastric administration of anthocyanins from R.Setate×R.Rugosa(high and low dose group),to observe the changes of the mice spleen index in mice,IL-2,IL-6 and TNF-alpha quantitative ELISA kit for determination of cellular immune function.Under the stimulation of Con A,the spleen cells were counted and the number of lymphocyte proliferation was detected by MTT.Research shows that: compared with the model group,anthocyanins from R.Setate×R.Rugosa in high dose group can increasethe spleen index in mice,increase spleen cell proliferation and in serum IL-2,reduced serum inflammatory cytokines IL-6 and TNF-alpha,statistical difference(P<0.05);The spleen index,serum IL-2 and spleen cell proliferation in mice with low dose of anthocyanins from R.Setate×R.Rugosa were not significantly increased,and the down regulation of IL-6 and TNF-alpha in the serum was not obvious.The results showed that anthocyanins in high dose group of bitter water rose can be used as an immunomodulator.