百里香酚类部位HPLC和UV-Vis法含量测定方法研究  

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

作　　者：范书生 隋宏[2] 陈晓怡[1] 王秀环 闫昕[1] 王乐 王小萍 李晓 许啸 李想 孙思琪 沈蒙[1] 折改梅[1] 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)

机构地区：[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

出　　处：《北京中医药大学学报》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)~~

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.

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 

分 类 号：R284.2[医药卫生—中药学]