淫羊藿不同部位多元活性成分的分析与评价  

Analysis and evaluation of bioactive constituents from different parts of Epimedium brevicornum

作　　者：薛佳 陈海杰 周永逸 袁嘉欢 蔡芷辰[1] 吴楠 陈翠花 刘训红[1,2] 邹立思 殷圣鑫[1] 杨薇 程建明 XUE Jia;CHEN Hai-jie;ZHOU Yong-yi;YUAN Jia-huan;CAI Zhi-chen;WU Nan;CHEN Cui-hua;LIU Xun-hong;ZOU Li-si;YIN Sheng-xin;YANG Wei;CHENG Jian-ming(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Province Engineering Research Center of Classical Prescription,Nanjing 210023,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]江苏省经典名方工程研究中心,江苏南京210023

出　　处：《中国中药杂志》2023年第13期3448-3461,共14页China Journal of Chinese Materia Medica

基　　金：江苏高校品牌专业建设工程项目(PPZY2015A070)。

摘　　要：建立超快速液相色谱-三重四极杆线性离子阱串联质谱(UFLC-QTRAP-MS/MS)同时测定淫羊藿黄酮、生物碱、氨基酸、酚酸、核苷类共45种活性成分含量的方法,并对淫羊藿叶片、叶柄、茎及根茎中多元活性成分进行分析与评价。采用XBridge C_(18)色谱柱(4.6 mm×100 mm,3.5μm),以0.4%甲酸水-乙腈为LC-MS系统流动相进行梯度洗脱,流速0.8 mL·min^(-1),柱温30℃;采用单因素试验及响应面法优化样品提取条件,系统聚类分析(SCA)、主成分分析(PCA)、偏最小二乘判别分析(PLS-DA)、单因素方差分析(One-way ANOVA)对不同部位样品进行区分并明确差异成分;采用灰色关联度分析(GRA)及熵权TOPSIS分析对不同部位样品进行综合排序。结果表明,45种目标成分的质量浓度与对应峰面积均呈现良好的对应关系(r≥0.999 0),精密度、重复性和稳定性良好,RSD均小于5.0%。平均加样回收率为94.51%~105.7%,RSD为0.62%~4.9%,以上结果表明淫羊藿不同部位多元活性成分具有一定差异,综合质量排序为叶片>叶柄>根茎>茎。验证了现行版《中国药典》规定淫羊藿药用部位为叶的合理性,同时发现淫羊藿根茎具有药用开发的潜力。该实验所建立的方法准确、可靠,可用于淫羊藿药材质量的综合评价和全面控制,同时为淫羊藿药用部位的确定以及合理利用淫羊藿资源提供实验依据。A comprehensive analytical method based on ultra-fast liquid chromatography coupled with triple quadrupole/linear ion trap tandem mass spectrometry(UFLC-QTRAP-MS/MS) was established for simultaneous determination of the content of 45 bioactive constituents including flavonoids,alkaloids,amino acids,phenolic acids,and nucleosides in Epimedium brevicornum.The multiple bioactive constituents in leaves,petioles,stems and rhizomes of E.brevicornum were analyzed.The gradient elution was performed at 30 ℃ in an XBridge C_(18) column(4.6 mm×100 mm,3.5 μm) with 0.4% formic acid aqueous solution-acetonitrile as the mobile phase at a flow rate of 0.8 mL·min^(-1).Single factor experiment and response surface methodology were employed to optimize the extraction conditions.Multivariate statistical analyses including systematic cluster analysis(SCA),principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA),and one-way analysis of variance(One-way ANOVA) were carried out to classify the samples from different parts and identify different constituents.Grey relation analysis(GRA) and entropy weight-TOPSIS analysis were performed to build a multi-index comprehensive evaluation model for different parts of E.brevicornum.The results showed that there was a good relationship between the mass concentrations of 45 constituents and the corresponding peak areas,with the correlation coefficients(r) not less than 0.999 0.The precision,repeatability,and stability of the established method were good for all the target constituents in this study,with the relative standard deviations(RSDs) less than 5.0%(0.62%-4.9%) and the average recovery of 94.51%-105.7%.The above results indicated that the bioactive constituents varied in different parts of E.brevicornum,and the overall quality followed the trend of leaves petioles rhizomes stems.This study verified the rationality of the Chinese Pharmacopoeia(2020 edition) stipulating that the medicinal part of E.brevicornum is the leaf.Moreover,our study indicated that the

关 键 词：淫羊藿 不同部位 超快速液相色谱-三重四极杆/线性离子阱质谱 多元活性成分 多元统计分析 

分 类 号：R284.1[医药卫生—中药学]