大孔树脂分离纯化红腰豆总黄酮工艺优化及体外抗氧化活性研究

来源 :中国农学通报 | 被引量 : 0次 | 上传用户:gustczh
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为研究大孔树脂纯化红腰豆黄酮的工艺和效果,以红腰豆总黄酮粗提液为原料,吸附率和解吸率为评价指标,比较8种树脂对红腰豆总黄酮的静态吸附和解吸性能,进行单因素试验和Box-Benhnken中心组合试验设计,运用响应面法分析统计优化AB-8型大孔树脂分离纯化红腰豆总黄酮的工艺条件,并研究了红腰豆总黄酮纯化前后体外抗氧效果。结果表明最佳纯化树脂为AB-8,最佳的吸附工艺条件为上样质量浓度4.0 mg/mL,上样液pH 6.3,上样流速2.0 mL/min,上样体积5.0 BV,在此条件下吸附率可达(98.027±0.303)%;最佳的解吸工艺条件为乙醇体积分数75%,洗脱流速3.0 mL/min,洗脱体积2.0 BV,在此条件下解吸率可达(94.523±0.240)%。纯化后红腰豆总黄酮纯度提高了约2.85倍,纯化前1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除率、羟基自由基(·OH)和超氧阴离子自由基(O2-·)的清除率IC50值分别为1.18、1.40、6.51 mg/mL,纯化后分别为0.37、0.82、1.77 mg/mL,纯化后红腰豆总黄酮提取物的体外抗氧化活性明显增强。“,”To study the process and results of macroporous resin purification of flavonoids from red kidney beans, the authors used the total crude extract from red kidney beans as raw material, compared eight kinds of resins’ static adsorption and desorption properties on the total flavonoids of red kidney beans with adsorption rate and desorption rate as the evaluation indexes. Single factor test and Box-Behnken central composite design were conducted with response surface methodology applied to analyze and optimize the conditions for AB-8 macroporous resin separation and purification of the total flavonoids from red kidney beans, and in vitro antioxidant effect before and after the purification of the total flavonoids was studied. The results showed that the optimum purification resin was AB-8. The best adsorption condition was 4.0 mg/mL sample concentration, pH 6.3 sample solution, 2.0 mL/min sample flow rate, 5.0 BV sample volume, under which, the adsorption rate could reach (98.027±0.303)%. Besides, the best desorption condition was 75% ethanol concentration, 3.0 mL/min elution flow rate, and 2.0 BV elution volume. And in this condition, the desorption rate could get to (94.523 ± 0.240) %. The total flavonoids purity of red kidney beans increased about 2.85 times after purification. Before purification, 1,1-diphenyl-2-trinitrobenzene hydrazine (DPPH) radical scavenging, hydroxyl radical (• OH) and superoxide anion radical (O2- •) clearance IC50 values were 1.18, 1.40, 6.51 mg/mL, respectively, which rose to 0.37, 0.82, 1.77 mg/mL after purification. After purification, in vitro antioxidant activity of the total flavonoids extract of red kidney beans was significantly enhanced.
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