The Optimum Extraction And Hypoglycemic Effect Of Anthocyanins In Black Soybean

Anthocyanin, a natural soluble food pigment, is widely distributed in nature because almost all of the higher plants can synthesize it. The most colors of red, blue, yellow and purple which fruits, flowers and vegetables are showing is because of anthocyanin, precisely because it was so natural showing a colorful scenery, recently, artificial coloring does so much harm to human body, World Health Organization prohibit the use of it, anthocyanin as a natural food coloring because of resources are abundant, safe, non-toxic, etc aroused great concern, and because it has some nutritional value and pharmacological, therefore, it has great prospects in food, pharmaceutical and cosmetic field.Black bean is rich in plant nutrients and is one of the health care quality goods, and it has special feature that is treatment with food, known as the "king of beans". It has planted in most parts of the country. Black bean red is distilled from black bean hull; anthocyanin is one of the main components. Black bean is inexpensive, readily available raw material; meet the needs of industrial production, therefore, in the areas of food and medicine will have great applications. In this paper, Luzhou black hull as the experimental material, studied the optimization of extraction of anthocyanin in pure ethanol extracted, assisted extraction of pectinase, assisted extraction of cellulose, then select the appropriate macroporous resin to purified, analysis on the black bean red anthocyanin stability factor, and the effect of black bean red anthocyanin act in lower the blood sugar was discussed. The results are as follows:(1) Put the black hull for the Material, made the extraction temperature(X₁), extraction time(X₂), ethanol concentration(X₃) and solid to liquid ratio(X4) of the four factors of the second general rotation four-factor design, the establishment of black bean red anthocyanin extraction conditions of mathematical regression model was: Y=1.18871+0.00288X₁+0.02987X₂-0.08929X₃+0.1517X₄-0.07758X₁²-0.00996X₂²- 0.16071X₃²-0.04396X₄²-0.05169X₁X₂+0.07756X₁X₃+0.05606X₁X₄+0.02969X₂X₃+ 0.0004X₂X₄-0.05381X₃X₄ Using the equation above, the best conditions of extracting black bean red anthocyanin are: solid to liquid ratio is 1:50g/mL, extraction time is 1 hour, Extraction temperature is 60℃, concentration of ethanol is 60%. The content of anthocyanin extracted under this optimal condition is 1.397mg / g.(2) Black bean red anthocyanin extracts after post-petroleum ether and ethyl acetate, the ability of water sector to clear DPPH is the best among ministry of petroleum ether, department of ethyl acetate and water sector. The suitable macroporous resin to purificate the anthocyanins of red beans was AB-8 macroporous resin, the absorption rate was 63.27%, Analysis was 82.24%; the optimal concentration of anthocyain was 6mg/5mL per 10mL AB-8 macroporous resin the optimal flow rate was 5mL/min, adsorption equilibrium time was 30mins, the optimal concentration of ethanol eluate was 70%. Finally set up a method to use the disodium hydrogen phosphate-sodium dihydrogen phosphate pH6.0 buffer, temperature 25℃, voltage 28KV, current 53.5μA, injection pressure 0.5psi, wavelength 220nm, the separation time of 10 minutes under conditions of capillary electrophoresis. The anthocyanin samples obtained by high performance capillary electrophoresis method should contain cyanidin -3-O- glucoside, and the content of cyanidin -3-O - glucoside was 0.454mg in 1.0g black bean hull.(3) By analyzing the impact of anthocyanin stability of red beans known that: Black bean red anthocyanin light resistance, heat resistance, resistance to oxidation are poor, should be saved in the dark, cold and sealed condition. Sodium chloride, vitamin c and glucose on the stability of anthocyanin in black bean red have no adverse effects, but the citric acid and sucrose was able to affect the stability of anthocyanin.(4) By different groups of normal mice and the use of alloxan modeling of experimental diabetic mice to the corresponding concentration of black beans were red anthocyanin intraperitoneal injection, and determination of glucose, superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), glutathione peroxidase (GSH-Px) and insulin levels. The results show that: black beans with red anthocyanin reduced the role of hyperglycemia in diabetic mice. The innovation of this experiment(1) Extracted from pure ethanol and pectinase assisted extraction of anthocyanin of red beans, black beans cellulase assisted extraction of anthocyanin in these three areas of red to red on black bean extract anthocyanin optimization study found that fruit gum and cellulose enzyme assisted extraction of anthocyanin of red beans and not simply the effect of acidified ethanol and good results.(2) Using high performance capillary electrophoresis (HPCE) on the black bean red anthocyanin quantitative analysis, using sodium hydrogen phosphate - sodium dihydrogen phosphate buffer pH6.0,temperature 25℃,voltage 28KV,current 53.5μA,injection pressure 0.5psi, wavelength 220nm, the separation time of 10 minutes under conditions of capillary electrophoresis analysis. At the same time, using the method to get the samples should contain anthocyanin cyanidin -3-O - glucopyranoside, and the black hull contains 1g of cyanidin -3-O - glucoside content of 0.454mg. Although the method still requires further study, since cyanidin -3-O - glucoside appeared two peaks.(3) The black bean red anthocyanin to hypoglycemic of mice were also studied, black beans and red anthocyanin had reduced role of hyperglycemia in diabetic mice, it can reduce the blood sugar and serum MDA and CAT, increase the serum SOD and GSH-Px levels ,also increase the serum levels of insulin.