| The black soybean, which is planted more and more now, is an important plant resource and existed in mountainous area in north of our country. It is widely used for the process of cosmetic and health care food,. It is reported that the anthocyanins in the coat of black soybean has the ability of antioxidant and scavenging hydroxyl radicals. This experiment used the coat of black soybean as material , in order to explored the process of extraction, purification and structural identification of the anthocyanins. All this done is aimed for providing theoretical basis and some scientific message for developing and utilizing black soybean resource.The methods that the dual wavelength spectrophotometry and HPLC, were established for the quantitative determination of the anthocyanins in the coat of black soybean. It is made certain by spectrum Scanning that the experimental conditions of dual wavelength spectrophotometry were as follow,detection wavelength 525nm. It is proved that this method was able to overcome the disturbance of the strong absorbance of solvent .HPLC was applied to quantitative determination of trigonelline in pumpkin flesh. The experimental conditions were as follows, the Agilent-1100 C18 column, mobile phase methanol: 0.4% phosphoric acid(55:45), flow rate 1.0ml/min, and detection Agilent- G1314A uv detector.In this experiment the process of anthocyanins extraction is explored. The anthocyanins in coat of black soybean was extracted by ultra sonic wave and alcohol extraction. The best method of alcohol extraction were obtained, namely extracted for 1h by using 60% ethanol solution at 60°C with the material ratio of 1:40(g/ml). Still studied the influence of the metals ion upon the black soybean skin.In this experiment, the macroporous adsorption resin and chromatographic methods were applied to the process of purification. The technologies were established by the choose of macroporous adsorption resin, the research of effects on adsorption and elution. The appropriate conditions were as follows, AB-8 macroporous adsorption resin, pH=6, elution solvent 60% alcohol solution, elution flow rate 2.5ml/min.The purified extraction was extracted by ethyl acetate. the ethyl acetate extraction was mixed with silica gel, dried in vacuum and encased in column. The elution solution was examined by paper chromatography. The hydrolysis part of the elution purified by silica gel was detected by HPLC.The Sephadex LH-20 column was used for purification of the hydrolysis part, 60% alcohol was chose as elution solvent, and elution flow rate was 1.0ml/min.. Lastly, we determined the purity of the compounds by melting point test, TLC and HPLC. The compounds were identified by UV and NMR methods. It is made certain that it is the Cyanidine-3-galactoside.The conclusion carried on the sour method and the enzyme method water solution to carry on a comparison to the anthocyanins for get, get is the enzyme method water to solve well, and to the cyanidine the research that carried on turning purely, get to turn purely of condition, use the HPLC and standard article ratio to, that material is cyanidine.It's still dye in the coat of black soybean carried on an anti- to oxidize to go of the research and other a few soybean skins carry on a comparison, its anti- oxidizes to go very good.
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