主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2019, Vol. 42 ›› Issue (10): 854-861.doi: 10.3969/j.issn.1006-2157.2019.10.010

• 中药化学 • 上一篇    下一篇

百里香酚类部位HPLC和UV-Vis法含量测定方法研究*

范书生1, 隋宏2, 陈晓怡1, 王秀环1, 闫昕1, 王乐1, 王小萍1, 李晓1, 许啸1, 李想1, 孙思琪1, 沈蒙1, 折改梅1#   

  1. 1 北京中医药大学中药学院 北京 102488;
    2 宁夏医科大学药学院
  • 收稿日期:2019-02-25 出版日期:2019-10-30 发布日期:2019-11-25
  • 通讯作者: 折改梅,女,博士,教授,博士生导师,研究方向:中(民族)药药效成分和新药创制研究,E-mail:shegaimei@126.com
  • 作者简介:范书生,男,在读硕士生
  • 基金资助:
    *国家自然科学基金资助项目(No.30020704),北京中医药大学青年教师项目(No. 1000061222118)

Determination of phenolic parts in ThymusquinquecostatusCelak by HPLC and UV-Vis*

Fan Shusheng1, Sui Hong2, Chen Xiaoyi1, Wang Xiuhuan1, Yan Xin1, Wang Le1, Wang Xiaoping1, Li Xiao1, Xu Xiao1, Li Xiang1, Sun Siqi1, Shen Meng1, She Gaimei1#   

  1. 1 School of Chinese Pharnacy, Beijing University of Chinese Medicine, Beijing 102488, China;
    2 School of Pharmacy, Ningxia Medical University, Yinchuan 750004, China
  • Received:2019-02-25 Online:2019-10-30 Published:2019-11-25
  • Contact: Prof. She Gaimei, Doctoral Supervisor. School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China. E-mail:shegaimei@126.com
  • Supported by:
    Natural Science Foundation of China(No.30020704), Young Teachers Program of Beijing University of Chinese Medicine(No. 1000061222118)

摘要: 目的 利用高效液相色谱法(HPLC)和紫外分光光度法(UV-Vis)建立民族药百里香酚类部位活性成分的含量测定方法。方法 以野芩苷和迷迭香酸为对照品,HPLC含量测定方法检测波长为280 nm,流动相为乙腈(A)-0.2%甲酸水(B)梯度洗脱,流速为1 mL/min,进样量为10 μL,柱温为25 ℃。UV-Vis含量测定方法选择三氯化铁-铁氰化钾显色方法,以迷迭香酸为对照品,检测波长为760 nm,显色剂用量为 2 mL,反应时间为 60 min。结果 野黄芩苷、迷迭香酸线性范围分别为0.019~0.592 g/L(R2=0.999)、0.009~0.280 g/L(R2=1.000),精密度(n=6)分别为0.21%、0.56%,稳定性RSD分别为0.30%、0.27%,重现性RSD分别为1.35%、1.47%,加样回收率分别为99.42%、95.31%、104.49%和101.23%、102.02%、97.62%,RSD分别为1.48%、1.46%、0.35%和0.29%、1.38%、2.05%;总多酚线性范围为1.10~2.05 mg/L(R2=0.999),精密度(n=6)RSD=0.97%,稳定性RSD=1.32%,重现性RSD=0.89%,加样回收率为100.88%,RSD为1.55%。测得三批酚类部位野黄芩苷和迷迭香酸平均含量分别为8.40%和8,23%;测得三批酚类部位总多酚平均含量为47.40%。结论 此两种方法简便准确,专属性强,灵敏度高,为百里香新药开发和后续研究奠定基础。

关键词: 百里香, 酚类部位, 高效液相色谱法, 紫外分光光度法, 含量测定

Abstract: ObjectiveTo establish methods for the determination of active components from phenolic parts of folk medicine Thymus quinquecostatus Celak by using HPLC and UV-Vis. Methods For HPLC, scutellarin and rosmarinic acid were used as references. The mobile phase A was acetonitrile while mobile B was 0.2% formic acid. The flow rate was set at 1.0 mL/min and the column temperature was maintained at 25 ℃ . 10 μL sample solutions were injected into auto sampler and monitored at 280 nm. For UV-Vis, developer dosage of K3[Fe(CN)6]-FeCl3 were chosen, rosmarinic acid was used as reference, the dosage of reagent was 2 mL, the determination time was 60 min and the detection wavelength was 760 nm. Results The linear ranges of scutellarin and rosmarinic acid were 0.019~0.592 g/L (R2=0.999), 0.999~0.280 g/L (R2=1.000), respectively. The RSDs(n=6) of precision experiment were 0.21%, 0.56%, respectively. The RSDs(n=6) of stability experiment were 0.21%, 0.56%, respectitvely. The RSDs(n=6) of reproducibility experiment were 1.35%, 1.47%, respectively, while the average adding standard recoveries were 99.42%, 95.31%, 104.49% (RSD=1.48%, 1.46%, 0.35%) and 101.23%, 102.02%, 97.62%(RSD=0.29%, 1.38%, 2.05%). The linear ranges of total polyphenolsisl 1.10~2.05 mg/L (R2=0.999). The RSD(n=6) of precision experiment was 0.97%. The RSD(n=6) of stability experiment was 1.32%. The RSD(n=6) of reproducibility experiment was 0.89%, while the average adding standard recoveries was 100.88%(RSD=1.55%). The average content of scutellarin and rosmarinic acid in all three batchesphenolic parts was 8.40% and 8.23%, respectively, while the averagecontent of total polyphenols was 47.40%.
Conclusion Both methods were simple, accurate, specific and sensitive, and can be foundations for the development and subsequent research of T. quinquecostatus Celak.

Key words: Thymus quinquecostatusCelak, phenolic parts, HPLC, UV-Vis, content determination

中图分类号: 

  • R284.2