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10μL,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(R^2=0.999)、0.009~0.280 g/L(R^2=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(R^2=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。
Objective To 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℃.10μL 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.
作者
范书生
隋宏
陈晓怡
王秀环
闫昕
王乐
王小萍
李晓
许啸
李想
孙思琪
沈蒙
折改梅
Fan Shusheng;Sui Hong;Chen Xiaoyi;Wang Xiuhuan;Yan Xin;Wang Le;Wang Xiaoping;Li Xiao;Xu Xiao;Li Xiang;Sun Siqi;Shen Meng;She Gaimei(School of Chinese Phamacy,Beijing University of Chinese Medicine,Beijing 102488,China;School of Pharmacy,Ningxia Medical University,Yinchuan 750004,China)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2019年第10期854-861,共8页
Journal of Beijing University of Traditional Chinese Medicine
基金
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)~~
