Extraction,Purification And Investigation Of Biological Activities Of Blueberry Anthocyanin

Blueberry is very popular as one of the five healthy foods acknowledged by Food and Agriculture Organization of the United Nations,for it is rich in anthocyanin.Since anthocyanin has many functions in health-care and pharmacology,the investigation of the extraction,purification and biological activities of blueberry anthocyanin is very significant in the application.Microwave-ethanol method was used to extract blueberry anthocyanin.With the appropriate conditions found through the single-factor experiments,the response surface method was used to adopt the quadratic polynomial mathematical model to optimize the process of blueberry anthocyanin extraction.Through the analysis of the effect to anthocyanin extraction rate with different extraction times,microwave powers,ethanol concentrations and liquid-solid ratios,by solving the corresponding regression equation,the optimal extraction conditions could be obtained under the following conditions:extraction time 2.87 min,microwave power 452 W,ethanol concentration 73%,liquid-solid ratio 22.With the chemical method,the purity of anthocyanin purified by different resins measured by chemical method showed that:NKA-9(71.62%),DM-7HP(74.62%),AB-8(62.61%),D101(41.59%),LKY-312(59.91%).With the HPLC method,the purity showed that:NKA-9(34.7%),DM-7HP(36.6%),AB-8(32.1%),D101(21.3%),LKY-312(28.2%).The chemical oxidation was used in the vitro experiment,and the method of establishing aging model and feeding different concentrations of blueberry anthocyanin extract was used in the vivo antioxidant experiment.The results showed that blueberry anthocyanin extract had a well scavenging effect on hydroxyl radicals,ABTS and DPPH,and well reducing power.In the vivo experiments,the blueberry anthocyanin extract could decrease the level of MDA in mice,and increase the level of SOD,CAT and GSH-PX in mice.Different methods were used to investigate the inhibition effect of blueberry anthocyanin extract on the proliferation of human hepatoma HepG-2 cells,such as the MTT method,the observation method with inverted microscope and the fluorescent staining method.The results showed that blueberry anthocyanin extract was good at the inhibition,the inhibition effects of 24 h and 48 h were respectively 38.39%and 48.52%.The observation method with inverted microscope and the fluorescent staining method could be used to observe the apoptosis of human hepatocellular carcinoma HepG-2 cells influenced by blueberry anthocyanin extract.The mice were used to establish the tumor model with S180 cells,fed with different concentrations of blueberry anthocyanin extract.Then these experiments were carried out to checkout the effect of the extract,such as the weight of the mice tumor,the spleen and thymus index of the mice,the level of TNF-?,TNF-? and TNFIL-II of the serum,the paraffin section experiment of the mice tumor.Otherwise,the mice with ascitic tumor were used to do the survival experiment.The results showed that blueberry anthocyanin extract had an obvious inhibition effect on the tumor in mice,the inhibition rate of the high dose group was 44.21%.At the same time,blueberry anthocyanin extract could enhance the level of spleen and thymus index,and increase the level of TNF-?,TNF-?and TNFIL-? in mice.Paraffin section observation of tumor tissue could indicate the characteristics of apoptosis and pathological characteristics of the tumor tissues.To a certain extent,blueberry anthocyanin extract could prolong the life of the mice.In conclusion,the anthocyanin extract,extracted and purified from blueberry,had good anti-tumor and anti-oxidant effects.