神农香菊茎叶总黄酮与总酚的提取纯化和抗氧化活性研究  

Extraction and Purification of Total Flavonoids and Total Phenols from Stems and Leaves of Chrysanthemum Indicum var. Aromaticum and Study of Antioxidant Activity

作　　者：冯颖 李彬 周春苗 叶利春 卢山 刘义飞 Feng Ying;Li Bin;Zhou Chunmiao;Ye Lichun;Lu Shan;Liu Yifei(College of Pharmacy,Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学药学院,武汉430065

出　　处：《世界科学技术-中医药现代化》2022年第4期1422-1432,共11页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：湖北省自然科学基金委员会重点类项目(2019CFA097):药用野菊的基因组遗传评价与优异种质筛选,负责人:刘义飞;湖北省财政厅、湖北省科学技术厅湖北省中央引导地方科技发展专项(2019ZYYD063):中药组学研究与资源评价创新平台,负责人:刘义飞。

摘　　要：目的 提高神农香菊茎叶总黄酮与总酚的产率和纯度并探究其抗氧化机制。方法 采用正交试验设计,优化神农香菊茎叶总黄酮和总酚的提取参数。使用大孔树脂吸附技术,对提取物中总黄酮和总酚进行纯化。通过UPLC-MS/MS分析纯化前后提取物中黄酮和酚酸类成分的含量变化。采用1,1-二苯基-2-苦肼基(DPPH)清除试验和分子对接评价提取物的抗氧化潜力。结果 最佳提取工艺为:料液比1:5 g·mL^(-1),乙醇浓度70%,提取时间60 min。最佳纯化条件为:上样浓度0.12 g·mL^(-1),上样体积3 BV,上样流速3.0BV/h,洗脱溶剂80%乙醇,洗脱流速4.5 BV/h,洗脱体积4 BV。与纯化前相比,纯化后提取物中总黄酮和总酚纯度分别提高了2.85倍和4.03倍。UPLC-MS/MS分析证实,纯化后部分黄酮和酚类物质含量显著增加。此外,神农香菊茎叶提取物显示出较强的DPPH自由基清除活性,其黄酮成分与黄嘌呤氧化酶具有较强结合能力。结论 本文对神农香菊茎叶总黄酮与总酚的富集和纯化具有重要指导意义,并表明了其潜在抗氧化价值。Objective To improve the yield and purity of total flavonoids and total phenols in stems and leaves of C.indicum var. aromaticum and explore their antioxidant mechanism.Methods The orthogonal experimental design was used to optimize the extraction parameters of total flavonoids and total phenols from stems and leaves of C. indicum var.aromaticum. The total flavonoids and total phenols in the extract were purified using macroporous resin adsorption technology. The contents of flavonoids and phenolic acids in the extracts before and after purification were analyzed by UPLC-MS/MS. The antioxidant potential of the extract was evaluated by 1, 1-diphenyl-2-picrohydrazine(DPPH)scavenging assay and molecular docking. Results The best extraction process was as follows: 1: 5 g·mL^(-1)ratio of material to liquid, 70% concentration of ethanol, and 60 min of extraction time. The best purification conditions were as follows: 0.12 g·mL^(-1)of sample concentration, 3 BV of sample volume, 3.0 BV/h sample flow rate, 80% ethanol of elution solvent, 4.5 BV/h of elution flow rate, and 4 BV of elution volume. Compared with before purification, the purity of total flavonoids and total phenols in the purified extract was increased by 2.85 times and 4.03 times, respectively. UPLC-MS/MS analysis confirmed that the contents of some flavonoids and phenolics were significantly increased after purification.In addition, C. indicum var. aromaticum stem and leaf extract showed strong DPPH free radical scavenging activity, and its flavonoids showed strong binding ability to xanthine oxidase. Conclusion This paper has important guiding significance for the enrichment and purification of total flavonoids and total phenols from stems and leaves of C. indicum var. aromaticum and shows their potential antioxidant value.

关 键 词：神农香菊 总黄酮 总酚 抗氧化活性 UPLC-MS/MS 分子对接 

分 类 号：R284.2[医药卫生—中药学]