| Health and security are always rewarded as important evaluation criterions for foods,drugs,and health products.Components with antioxidative bioactivities from natural plants,such as functional oils,flavonoids,polysaccharides and anthocyanins have obtained extensive attention and application in food,medicine and cosmetics industry contributing to their relatively high safety of use.Black soybean(Glyeine Soja Sieb.et.Zucc)as seeds for legume soybean plants is also known as Wudou,and classified to green core black soybean and yellow core black soybean according to the core colour.We have abundant germplasm resources of soybean in our country which is vast in territory and resulted in various kinds of black soybean with distinct ingredients.Xiaogang black soybean originated from He country,Sixian country and Fengyang country in Anhui province is famous for the abundance of protein,fat and other nutritions.However,the composition has not been systematically studied,much less the functions of individual component.So that the premium resource can't been effectively developed and utilized.The present study aimed at problems during the development of Xiaogang black soybean expecting to provide theoretical bases for continual exploitation.The basic compositions including water,protains,amino acids,crude fat and ash in Xiaogang black soybean were all determined by national standards.Part of the antioxidants in Xiaogang black soybean including ?-linolenic acid,?-linolenic acid and total flavones were detected by capillary gas chromatography or other methods,and the anoxidative activities of different types black soybean were compared by DPPH and FRAP methods.The optimization of Xiaogang black soybean polysaccharides extraction was studied by response surface methodology in an ultrasonic alkaline solution.Meanwhile,the optional pigment absorbent material for Xiaogang black boybesan was decided and the static adsorption,dynamic absorption and desorption process conditions were subsequently optimized.HPLC anylasis was used to separate the pigment and HPLC-MS to identify the anthocyanins.Results are as follows:1.The compositions of XiaoGang black soybean seed:(1)The XiaoGang Black Soybean contained 7.0±0.1% water,18.31±0.1% fat,37.38±0.11% protein,1.77±0.02% ash and 1.77±0.02% reducing sugar;(2)Xiaogang Black Soybean contains resourceful essential amino acids including 1.2mg/100 mg isoleucine,2.19 mg / 100 mg Leucine,1.87 mg / 100 mg Lysine,0.37 mg /100 mg Methionine,1.39 mg / 100 mg pHenylalanine,1.09 mg / 100 mg Threonine,1.18 mg/ 100 mg Valine and 0.75 mg / 100 mg Histidine.(3)772.1 mg/100 g Calcium,22.98 mg/100 g Iron,6.16 mg/100 g Copper,8309.42mg/100 g Potassium,22.94 mg/100 g Manganese,24.92 mg/100 g Zinc were also implicated in Xiaogang Black Soybean.2.The concentration of ?-linolenic acid and ?-linolenic acid were simultaneously determined by a capillary gas chromatograpHic method.The ?-linolenic acid and?-linolenic acid esters are completely separated through DB-WAX capillary column(30m×0.32 mm×0.25 ?m)and resolution >2.Using heptadecanoic acid methyl ester being as internal standard substance,?-linolenic acid and ?-linolenic acid showed good linear relations(r>0.999)between peak area and mass concentration.The recoveries for the spiked standards and the relative standard deviations are 87.5%~95% and 4.8%~0.9%,respectively,and the detection limit(s/n=3)is 0.0002 mg/mL.Given the mentioned results,we selected four representative black soybean in the domestic market to determine the concentration of ?-linolenic acid(8070~33000 mg/kg)and ?-linolenic acid(0 ~770 mg/kg).3.5.9% total flavonoids was found in Xiaogang Black soybean.To investigate the relationship between the total flavons and antioxidative activities,ethanol extractions of four black soybeans was used to detect this two indicators.The DPPH/ FRAP correlation coefficient was 0.8982> 0.8,both of which showed good correlations,indicating the effects of flavons against oxidation.4.In order to explore the optimal conditions of crude polysaccharides extraction in ultrasonic assisted alkali solution,the response surface analysis were used on the base of single factor experiment.The optimum condition was composed an alkali concentration of0.35 mol/L,an ultrasonic time of 16 min under 90 ?,with a 7.415% crude polysaccharide of polysaccharides.5.Among the tested resin,HP-20 macroporous resin purification pigment black hull optimum adsorbent.The biggest adsorptive capacity of HP-20 macroporous resin was12.157 mg/g wet resin,the rate of desorption respectively was 87.16%.When the pH value of adsorption fluid was 2-3,the A was1.084,the elution liquid was 60% ethanol(pH=2.2)and,the speed of adding was 1.5L/min and the elution flow rate was 2m L/min which,the separate efficacity of the Xiaogang black soybean.The color value of the pigment extrations performed 2.5 times higher than initial liquid after purification.6.After series of column chromatograpHy(HP-20,polyamide),three kinds of pigment anthocyanins were isolated from Xiaogang Black soybean,which can be primarilyidentified as Cyanidin-3-O-glucoside chloride,Delphinidin-3-glucoside and Petunia3-glucoside by HPLC-MS. |
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