Study On The Properties Of Anthocyanins From Black Soybean Coats And Inhibition Of The Proliferation Of Tumor Cells

Black soybean is a kind of edible and medicinal leguminous plant and the peel of black soybean is rich in phenols which has been shown in some researches also make it an important source of natural pigment. Because of its safety and non-toxic characteristics, natural pigment has a high development and utilization value and a wide development and application prospect.The research applied with water bath extraction method and microwave assisted extraction method for extracting anthocyanin from black soybean seed coats and we calculated and compared the rate of anthocyanin extraction and yield. The results showed that the microwave assisted extraction method is better than water bath assisted extraction and the optimum extraction conditions of microwave assisted extraction as following:the ratio of material to liquid is 1:30, microwave power is 500 W and the extraction agent is 60% ethanol and pH is 2. Meanwhile, we identified the anthocyanins samples using high performance liquid chromatography after the purification with the D101 macroporous resin. The chromatograms showed that the cyanidin-3-O-glucoside is the main component of the anthocyanins from the black soybean seed coats. Moreover, the purity of sample was improved about 1.14 times after purified. Combined with the visible spectrophotometry method, we analysised some common factors that influence the stability of the anthocyanin. Light and temperature had less effect on the anthocyanin. Potassium sorbate of preservative and sodium benzoate affect the stability of anthocyanin in a dose-dependent manner. Effects of Na+ and K+ on the stability of anthocyanin were not significant. Mg2+ has a mild color protecting effect on the anthocyanin. Ca2+ has a little effect on the anthocyanin. Effect of Fe3+ on the stability of anthocyanin is significant. Sugar almost does not affect its stability. However, the stability of anthocyanin is severely damage in the presence of redox agent.The results of MTT assay indicate that the anthocyanin from black soybean seed coats can effectively inhibit the proliferation of MCF-7, HCT-116 and HepG2. In particular, the inhibitory effect of anthocyanin on HepG2 cells was most significant. Anthocyanin doesn’t affect the normal intestinal cell FHC’s proliferation when the sample concentrations is below 250 μg/mL. The determination results of CAA value, SOD activity and CAT activity indicated that the anthocyanin from black soybean seed coats has some oxidation resistance in a dose-dependent manner.The results of this study will find the potential application of anthocyanin from black soybean seed coats on the anti-tumor field, and lay the theoretical basis for the development of health product or medicine with anti-tumor activity.