生物技术通报 ›› 2017, Vol. 33 ›› Issue (5): 63-70.doi: 10.13560/j.cnki.biotech.bull.1985.2017.05.009

• 技术与方法 • 上一篇    下一篇

沙冬青种子总黄酮测定方法及纯化工艺研究

陶波,方梅,张嘉男,欧云文,贾宁   

  1. 甘肃农业大学动物医学院,兰州 730070
  • 收稿日期:2016-10-30 出版日期:2017-05-25 发布日期:2017-05-19
  • 作者简介:陶波,男,硕士研究生,研究方向:动物病理学及新兽药研发;E-mail:315661125@qq.com
  • 基金资助:
    国家自然科学基金项目(31560686)

Determination and Purification of Total Flavonoids from Seeds of Ammopiptanthus mongolica

TAO Bo FANG Mei ZHANG Jia-nan OU Yun-wen JIA Ning   

  1. College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070
  • Received:2016-10-30 Published:2017-05-25 Online:2017-05-19

摘要: 旨在建立沙冬青种子总黄酮测定方法及沙冬青种子总黄酮纯化工艺。选用常用4种黄酮显色方法,通过紫外波长扫描分别确定槲皮素为对照品及沙冬青种子总黄酮的最佳吸收波长,建立标准曲线,并对各测定方法进行方法学验证,利用新建立的条件方法对沙冬青种子总黄酮进行含量测定;实验选择AB-8、D101、S-8型大孔树脂通过静态吸附及解析试验,筛选出AB-8型大孔树脂进行动态试验,建立大孔树脂纯化工艺条件,进一步通过乙酸乙酯进行萃取,提高其总黄酮纯度。结果显示,以槲皮素为对照品,采用AlCl3-CH4O显色法,在检测波长为335 nm条件下可对沙冬青种子总黄酮含量进行有效测定;AB-8型大孔树脂在上样液浓度为6 mg/mL、上样液pH5.5、上样液体积5 BV、上样液流速3.5 BV/h条件下吸附,以70%乙醇、洗脱液流速1.5 BV/h、洗脱液体积6 BV条件下洗脱,沙冬青种子总黄酮含量由纯化前5.69%提高到41.07%;用AB-8树脂纯化后的总黄酮配置成一定浓度溶液后用乙酸乙酯萃取,总黄酮含量达到76.15%。

关键词: 沙冬青种子, 总黄酮, AlCl3显色法, 大孔树脂, 萃取, 纯化

Abstract: This work is to establish the determination and purification method of total flavonoids from seeds of Ammopiptanthus mongolica. Four chromogenic methods were chosen for the determination of total flavonoids,the best absorption wavelengths were selected by UV-wavelength scanning of quercetin standard and total flavonoids from seeds of A. mongolica. Standard curves for quercetin by four chromogenic methods were built,and each method was verified via methodology,and the new experimental condition and method after verification were used to determine the content of total flavonoids from seeds of A. mongolica. In this experiment,3 kinds of macroporous resins of AB-8,D101,and S-8 were selected for static adsorption and elution test,and AB-8 was chosen for dynamic test,with which the process condition for purifying total flavonoids from seeds of A. mongolica was established,and the extraction was conducted with ethyl acetate to improve the purity of total flavonoid. The results showed that the determination of total flavonoids from seeds of A. mongolica with AlCl3-CH4O chromogenic method at 335 nm and quercetin as the standard substance was reliable and effective. Under the optimal condition,adsorption with AB-8 macroporous resins was 5 BV of 6 mg/mL sample solution at pH 5.5 at rate of 3.5 BV/h,and then elution was 6 BV of 70% ethanol at rate of 1.5 BV/h,the purity of the product was 41.07% from 5.69% before purification. The total flavonoids after purified with AB-8 resin were prepared into a certain concentration solution,and then extracted with ethyl acetate,the total flavonoids content reached 76.15%.

Key words: seeds of Ammopiptanthus mongolica, total flavonoids, AlCl3 chromogenic method, macroporous resin, extraction, purification