黑豆异黄酮的提取分离及其对DPPH自由基的清除能力  

Extraction of Isoflavones from Glycine max and Its Ability to Scavenge DPPH Free Radicals

作　　者：王宁[1] 张叶韬 芦晓芳[1] WANG Ning;ZHANG Ye-tao;LU Xiao-fang(Foundation Department,Shanxi Agricultural University,Taigu,Shanxi 030801)

机构地区：[1]山西农业大学基础部,山西太谷030801

出　　处：《安徽农业科学》2022年第1期171-176,共6页Journal of Anhui Agricultural Sciences

基　　金：山西省高等学校大学生创新创业训练项目(202010113014);山西农业大学博科研启动(J242098140)。

摘　　要：基于超声时间、液料比和超声温度的单因素试验,通过响应面分析法优化,结合样品粉末的SEM表征,确定超声辅助法提取黑豆异黄酮的最优工艺条件;经大孔树脂(D-101)纯化黑豆异黄酮粗提物,继而以乙酸乙酯-甲醇(V乙酸乙酯∶V甲醇=10∶1)溶液洗脱过硅胶层析柱,得组分Ⅰ和组分Ⅱ;考察异黄酮、组分Ⅰ和组分Ⅱ体外抗氧化活性。结果表明:超声辅助法提取黑豆异黄酮的最优工艺参数为时间38 min,液料比32∶1,温度53℃,该条件下黑豆异黄酮提取量[(9.403±0.698)mg/g]最大;经大孔树脂(D-101)纯化得到纯度较高(异黄酮含量[(53.37±3.37)%]的异黄酮;过硅胶层析柱得组分Ⅰ和组分Ⅱ。当总异黄酮、组分Ⅰ和组分Ⅱ的浓度达到0.25 mg/mL时,DPPH·自由基清除率分别达到了(71.28±1.41)%、(34.60±2.95)%和(42.01±1.34)%,总异黄酮的清除率是组分Ⅰ和组分Ⅱ的2.06和1.70倍,总异黄酮、组分Ⅰ和组分Ⅱ的抗氧化能力依次为总异黄酮>组分Ⅱ>组分Ⅰ。The optimal process conditions for ultrasonic-assisted extraction of isoflavones from Glycine max were determined based on univariate analysis of ultrasonic time,solid-liquid ratio and ultrasonic temperature,and SEM characterization of sample powder.After purifying the crude extract of isoflavones from Glycine max by the macroporous resin(D-101),and silicagel column chromatographic separation in ethyl acetate-methanol(Vethyl acetate∶Vmethanol=10∶1),component I and component II were obtained.The in vitro anti-oxidant activities of isoflavones,component I and component II were analyzed.The results showed that the optimal process parameters for the extraction of isoflavones from Glycine max by the ultrasonic-assisted method were as follows:time,38 min;liquid-solid ratio,32∶1;temperature,53℃.Under these conditions,the extraction rate of isoflavones from Glycine max achieved the highest(9.403±0.698)mg/g.Using the macroporous resin(D-101),highly purified isoflavones were extracted from Glycine max(53.37±3.37)%.Component I and component II were obtained from the silicagel column chromatographic separation.Under the concentrations of total isoflavones,component I and component II at 0.25 mg/mL,their DPPH free radical scavenging rate reached(71.28±1.41)%,(34.60±2.95)%and(42.01±1.34)%,respectively.In particular,the scavenging rate of total isoflavones was 2.06 and 1.70 times than that of component I and component II.The antioxidant capacity remained highest in total isoflavones,followed by component II,and component I.

关 键 词：响应面法 异黄酮 分离纯化 抗氧化活性 

分 类 号：TQ914.1[化学工程]