AB-8大孔树脂对红腰豆总黄酮纯化工艺的优化及其体外抗氧化活性研究

以红腰豆总黄酮粗提液为原料,研究大孔树脂对红腰豆黄酮的纯化工艺和效果,比较了8种树脂对红腰豆总黄酮的静态吸附和解吸性能,对AB-8型大孔树脂分离纯化红腰豆总黄酮进行了单因素、Box-Benhnken中心组合设计和响应面法优化试验,并考察了红腰豆总黄酮纯化前后体外抗氧效果。结果表明,AB-8树脂为纯化红腰豆总黄酮的最佳树脂,确定了其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(98.03±0.30)%；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(94.52±0.24)%。纯化后红腰豆总黄酮纯度提高了约2.85倍,纯化前DPPH.、.OH和O_2-.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。

Optimization of AB-8 Macroporous Resin to Purification of Total Flavonoids From Red Kidney Beans and In Vitro Antioxidant Activity

The process and results of macroporous resins for the purification of flavonoids from red kidney beans were studied, the performances of adsorption-desorption of the eight kinds of resin were investigated by using adsorption rate and desorption rate as 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 total flavonoids from red kidney beans, and studied vitro antioxidant effect before and after the purification of red kidney beans total flavonoids. The results showed that the optimum purification resin was AB-8. The best adsorption conditions were 4.0 mg/mL sample concentration, pH 6.3, loading velocity 2.0 mL/min, sample volume 5.0 BV, under which condition, the adsorption rate could reach (98.03±0.30)%. Besides, the best desorption conditions were 75% ethanol concentration, desorption velocity 3.0 mL/min and eluant volume 2.0 BV. And in this condition, desorption rate could get to (94.52 ± 0.24) %. 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 (O_2- ?) clearance IC₅₀ values were 1.18, 1.40, 6.51 mg/mL, respectively, which increased to 0.37, 0.82, 1.77 mg/mL after purification. After purification, in vitro antioxidant activity of red kidney bean total flavonoids extract significantly enhanced.