目的 研究大孔树脂纯化驱虫斑鸠菊总黄酮的工艺和体外抗氧化活性。方法 通过静态吸附解吸实验确定大孔树脂选型,研究其吸附动力学、热力学。通过单因素考察该树脂对总黄酮纯化工艺的影响。在此基础上,以总黄酮回收率为指标,用U10(103)均匀设计法优化并确定最佳工艺。考察纯化前后总黄酮体外抗氧化活性。结果 NKA-9型树脂对驱虫斑鸠菊总黄酮吸附分离性能良好,符合拟一阶动力学模型和Langmuir热力学模型。最佳工艺为:上样液浓度1.0 mg·mL-1,调pH 0.5,最大上样量2.3倍柱体积,除杂溶剂pH≤2,洗脱液为体积分数75%的乙醇,洗脱液体积6倍柱体积,洗脱流速1 mL·min-1。在该条件下,总黄酮回收率为98.19%,放大试验显示总黄酮回收率为94.37%,含量为81.02%,对1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-联氮-二(3-乙基-苯并噻唑-b磺酸)二铵盐(ABTS)自由基清除率分别为83.76%和92.64%。结论 优选的纯化工艺稳定可行,适于驱虫斑鸠菊总黄酮的纯化。
Abstract
OBJECTIVE To investigate the purification process and in vitro antioxidant activity of total flavonoids from Vernonia anthelmintica by macroporous resin. METHODS The selection of macroporous resin was determined by static adsorption and desorption experiments, which the adsorption kinetics and thermodynamics were studied. The effects of the selected resin on the purification process of total flavonoids were investigated by single factor experiments. On this basis,the U10(103) uniform design was used to optimize and determine the best purification process using the total flavonoid recovery as an index. The antioxidant activity of total flavonoids before and after purification was examined. RESULTS NKA-9 resin showed good adsorption and separation property for Vernonia anthelmintica flavonoids, which consistent with the Quasi first order model and the Langmuir isotherm adsorption equation. Optimal processes are as follows: sample concentration was 1.0 mg·mL-1, pH was adjusted to 0.5, the largest sample load was 2.3 times column volume, solvent which pH≤2 was used for impurity removal, eluent concentration was 75% ethanol solution and the dosage was 6 times column volume with elution rate of 1 mL·min-1. Under these conditions, the total flavonoids recovery was 98.19%, while scale-up test shown was 94.37%, the content was 81.02%, and the scavenging of DPPH and ABTS radicals was 83.76% and 92.64%, respectively. CONCLUSIONS The optimized purification technology is stable, feasible and suitable for the purification of total flavonoids from Vernonia anthelmintica.
关键词
大孔树脂 /
均匀设计法 /
驱虫斑鸠菊总黄酮 /
纯化工艺 /
抗氧化活性
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Key words
microporous resin /
uniform design /
Vernonia anthelmintica total flavonoid /
purification process /
antioxidant activity
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中图分类号：
R282
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参考文献
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脚注
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基金
国家自然科学基金项目资助(81860730);新疆生产建设兵团重大科技项目资助(2020AA005)
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