Page 69 - 202007

草果总黄酮的大孔吸附树脂纯化工艺优化研究 Δ

1
1，2 #
1，2
1
1
1＊
1
赵雨鸿 ，沈 华 ，代双亿 ，蒲忠慧 ，谢子锐 ，肖 灵 ，况 敏 ，代 敏 （1.成都医学院检验医学院，成都
1
610500；2.成都医学院四川省动物源性食品兽药残留防控技术工程实验室，成都 610500）
中图分类号 R284.2 文献标志码 A 文章编号 1001-0408（2020）07-0831-06
DOI 10.6039/j.issn.1001-0408.2020.07.13
摘 要 目的：建立草果总黄酮的含量测定方法，并对其大孔吸附树脂纯化工艺进行优化。方法：采用高效液相色谱法测定草果
中总黄酮的含量。色谱柱为Eclipse Plus C18，流动相为乙腈-1％醋酸水溶液（15∶85，V/V），柱温为40 ℃，流速为0.8 mL/min，检测波
长为256 nm，进样量为10 μL。以吸附、解吸性能为考察指标，采用静态吸附和解吸试验对6种大孔吸附树脂进行筛选，采用静态
吸附和解吸动力学试验考察吸附和解吸时间。在单因素试验的基础上，以总黄酮含量（以芦丁计）为评价指标，以上样液质量浓
度、上样液pH、乙醇体积分数及洗脱用量为考察因素，采用正交设计优化草果总黄酮的纯化工艺并进行验证试验。结果：芦丁检
测质量浓度的线性范围为0.028～0.281 mg/mL（r＝0.999 9）；定量限为437.5 ng/mL，检测限为109.4 ng/mL；精密度、稳定性、重复
性试验的RSD均小于2％，加样回收率为96.24％～99.75％（RSD＜2％，n＝6）。HPD450型大孔吸附树脂对草果总黄酮的静态吸
附和解吸综合能力最适中，最佳静态吸附与解吸时间均为12 h。最优纯化工艺为上样质量浓度1.854 4 mg/mL，上样液pH 7，乙醇
体积分数60％，乙醇洗脱用量8倍柱体积。验证试验显示，按最优工艺纯化后的草果总黄酮含量由纯化前的22.556 7 mg/g上升至
57.728 2 mg/g，纯化倍数为2.56（n＝3）。结论：所建含量测定方法灵敏度高、稳定性好，优化的纯化工艺稳定、可行，可用于草果总
黄酮的纯化。
关键词 草果；总黄酮；含量测定；大孔吸附树脂；纯化工艺

Study on the Optimization of Purification Technology of Total Flavonoids from Amomum tsao-ko by
Macroporous Adsorption Resin
ZHAO Yuhong ，SHEN Hua ，DAI Shuangyi ，PU Zhonghui ，XIE Zirui ，XIAO Ling ，KUANG Min ，DAI Min 1，2
1
1
1
1
1
1
1，2
（1. School of Laboratory Medicine，Chengdu Medical College，Chengdu 610500，China；2. Sichuan Provincial
Engineering Laboratory for Prevention and Control Technology of Veterinary Drug Residue in Animal-origin
Food，Chengdu Medical College，Chengdu 610500，China）
ABSTRACT OBJECTIVE：To establish a method for the content determination of total flavonoids from Amomum tsao-ko，and to
optimize the purification technology by macroporous resin. METHODS：The content of total flavonoids was measured by HPLC.
The determination was performed on Eclipse Plus C18 column with mobile phase consisted of acetonitrile-1％ acetic acid solution
（15 ∶ 85，V/V）at the flow rate of 0.8 mL/min. The column temperature was 40 ℃，and the detection wavelength was set at 256 nm.
The sample size was 10 μL. Taking the adsorption and desorption performance as indexes，6 kinds of macroporous resins were
screened out by static adsorption and desorption tests；adsorption and desorption time were investigated by static adsorption and
desorption kinetics tests. Using the content of total flavonoids（calculated by rutin）as index，with sample concentration，sample
pH，ethanol volume fraction and elution amount as factors，based on single factor test，orthogonal design was used to optimize the
purification technology of total flavonoids from A. tsao-ko，and validation test was performed. RESULTS：The linear range of rutin
were 0.028-0.281 mg/mL（r＝0.999 9）. The limit of quantification was 437.5 ng/mL and the limit of detection was 109.4 ng/mL.
RSDs of precision，stability and reproducibility tests were all lower than 2％；the recoveries were 96.24％-99.75％（RSD＜2％，n＝
6）. The comprehensive capacity of adsorption and desorption of HPD450 macroporous resin was the most suitable，and the best
static adsorption and desorption time both were 12 h. The optimal purification technology was 1.854 4 mg/mL，sample pH value
was 7，ethanol volume fraction was 60％ ，the amount of
Δ 基金项目：四川省科技创新苗子工程资助项目（No.2018086）；
四川省省级大学生创新创业训练计划项目（No.201913705037）；成都 ethanol elution was 8 times of the column volume. Vertification
医学院校基金计划（No.CYZ18-18）；成都医学院检验医学院院级自然 tests show that after optimized，the content of total flavonoids
科学研究项目（No.JYZK201802） from A. tsao-ko increased from 22.556 7 mg/g to 57.728 2
＊本科生。研究方向：天然药物提取、分离与纯化。E-mail： mg/g. The purity of was 2.56 times higher than before
1546129084@qq.com
purification. CONCLUSIONS：Established method is sensitive
# 通信作者：讲师。研究方向：天然药物化学成分及药效物质基
and stable for the content determination. Optimal purification
础。E-mail：zhonghui.pu@163.com

中国药房 2020年第31卷第7期 China Pharmacy 2020 Vol. 31 No. 7 ·831 ·