摘要
目的:建立刺五加的超高效液相色谱法(UPLC)多成分含量测定与指纹图谱方法,对34批不同生长条件、不同药用部位刺五加进行含量测定、指纹图谱分析和化学模式识别。方法:以Waters ACQUITY UPLC CORTECS T3色谱柱(150 mm×2.1 mm,1.6μm)为固定相,0.1%甲酸水溶液(A)-0.1%甲酸乙腈溶液(B)为流动相梯度洗脱;流速为0.3 mL·min^(-1),检测波长为265 nm,柱温为30℃,进样量为1μL,测定10个化合物(原儿茶酸、紫丁香苷、刺五加苷B1、刺五加苷E、异嗪皮啶、金丝桃苷、绿原酸、异绿原酸A、异绿原酸B、异绿原酸C)的含量;采用指纹图谱分析识别共有峰并计算相似度;运用系统聚类分析(HCA)、主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)研究样品的聚类规律并筛选差异标志成分。结果:建立了刺五加UPLC多成分含量测定和指纹图谱方法。含量测定结果显示,10个化合物在不同生长条件、不同药用部位刺五加中的含量存在一定差异,其中绿原酸、原儿茶酸、金丝桃苷、总香豆素、总异绿原酸的含量在刺五加茎、根及根茎之间差异有统计学意义(P<0.05)。34批刺五加指纹图谱共标定28个共有峰;17批刺五加茎指纹图谱共标定33个共有峰;17批刺五加根及根茎指纹图谱共标定40个共有峰。HCA、PCA可以较好地区分刺五加茎、根及根茎,表明刺五加茎、根及根茎之间存在较大的差异。OPLS-DA表明原儿茶酸、绿原酸、异绿原酸A、异绿原酸C可能是造成刺五加质量差异的标志物。结论:建立的UPLC多成分含量测定方法和指纹图谱法适用于不同生长条件、不同药用部位刺五加的质量评价;相关的研究结果可为刺五加的质量控制提供科学的参考依据。
Objective:To establishing a ultra-high performance liquid chromatography(UPLC)-based multi-component content determination and fingerprinting method for Acanthopanax senticosus,and apply this method for content determination,fingerprinting,and chemical pattern recognition for 34 batches of A.senticosus samples from different growth conditions and different medicinal parts.Methods:Gradient elution was performed with Waters ACQUITY UPLC CORTECS T3 column(150 mm×2.1 mm,1.6μm)as the stationary phase and 0.1% formic acid water(A)-0.1% formic acid acetonitrile(B)as the mobile phase,with a flow rate of 0.3 mL·min^(-1).The detection wavelength,column temperature,and injection volume were 265 nm,30℃,and 1μL,respectively.The content of 10 compounds such as protocatechuic acid,eugenin,eleutheroside B1,eleutheroside E,isozinepicidine,hypericin,chlorogenic acid,isochlorogenic acid A,isochlorogenic acid B,and isochlorogenic acid C in the A.senticosus samples was determined.The common peaks in fingerprints were identified and the similarity was calculated.Hierarchical cluster analysis(HCA),principal component analysis(PCA),and orthogonal partial least squares discriminant analysis(OPLS-DA)were performed to study the clustering patterns and screen differential marker components.Results:A UPLC-based multi-component content determination and fingerprinting method was established for A.senticosus.The content of 10 compounds varied among the A.senticosus samples from different growth conditions and different medicinal parts.The content of chlorogenic acid,protocatechuic acid,hyperoside,total coumarin,and total isochlorogenic acid showed differences between stem samples and root&rhizome samples of A.senticosus(P<0.05).A total of 28 common peaks were identified in the fingerprints of 34 batches of A.senticosus;33 common peaks were identified in the fingerprints of 17 batches of A.senticosus stem samples;and 40 common peaks were identified in the fingerprints of 17 batches of A.senticosus root&rhizome samples.HCA and PCA distinguished the stem samples and the root&rhizome samples of A.senticosus,indicating a significant difference between the two medicinal parts.OPLS-DA indicated that protocatechuic acid,chlorogenic acid,isochlorogenic acid A,and isochlorogenic acid C may be markers responsible for the quality differences of A.senticosus samples.Conclusion:The established UPLC-based multi-component content determination and fingerprinting method is suitable for the quality evaluation of A.senticosus samples from different growth conditions and different medicinal parts.The research results can provide scientific references for the quality control of A.senticosus.
作者
黄鹏程
冯龙峰
刘彦兵
吴宙
柯潇
肖强
王秋玲
魏建和
HUANG Pengcheng;FENG Longfeng;LIU Yanbing;WU Zhou;KE Xiao;XIAO Qiang;WANG Qiuling;WEI Jianhe(Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100193,China;Sichuan Kanghong Chinese Herbal Medicine Planting Co.,Ltd.,Chengdu 611930,China;Hainan Branch of Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Haikou 570311,China)
出处
《中国现代中药》
2025年第9期1683-1695,共13页
Modern Chinese Medicine
基金
高品质中药材生态调控栽培技术研究项目(2022YFC3501500)
中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-032)
国家中医药管理局项目(GZY-KJS-2023-008)。
关键词
刺五加
超高效液相色谱法
多指标含量测定
指纹图谱
化学识别模式
Acanthopanax senticosus(Rupr.ex Maxim.)Harms
UPLC
content determination
fingerprint
chemical pattern recognition
