茶芎HPLC指纹图谱的建立及差异性成分的含量测定
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篇名: 茶芎HPLC指纹图谱的建立及差异性成分的含量测定
TITLE: Establishment of HPLC fingerprint and content determination of differential components in Ligusticum sinense
摘要: 目的 建立不同产地茶芎的指纹图谱,筛选差异性成分并测定其含量。方法以Z-藁本内酯为参照,采用《中药色谱指纹图谱相似度评价系统(2012版)》建立12批茶芎药材的高效液相色谱(HPLC)指纹图谱,指认共有峰并进行相似度评价;进行聚类分析(CA)、主成分分析(PCA)和正交偏最小二乘-判别分析(OPLS-DA),以变量重要性投影(VIP)>1为标准筛选影响药材质量的差异性成分;采用同一HPLC法测定上述差异性成分的含量。结果12批茶芎药材指纹图谱共有17个共有峰,相似度为0.989~1.000;共指认了7个共有峰,分别为绿原酸(1号峰)、阿魏酸(2号峰)、洋川芎内酯Ⅰ(7号峰)、阿魏酸松柏酯(9号峰)、E-藁本内酯(13号峰)、洋川芎内酯A(14号峰)、Z-藁本内酯(17号峰)。CA结果显示,12批茶芎药材可分为3类,S1~S5(武宁茶芎)聚为一类,S6~S8(瑞昌茶芎)聚为一类,S9~S12(德安茶芎)聚为一类;2、13、14、17号峰(分别对应阿魏酸、E-藁本内酯、洋川芎内酯A、Z-藁本内酯)的VIP均大于1。S1~S5、S6~S8、S9~S12样品中,阿魏酸的含量分别为0.488~0.533、0.603~0.658、0.415~0.433mg/g,洋川芎内酯A分别为1.184~1.295、1.450~1.588、1.307~1.377mg/g,E-藁本内酯分别为0.118~0.125、0.130~0.135、0.223~0.229mg/g,Z-藁本内酯分别为7.200~7.681、8.076~8.643、4.508~4.996mg/g,两两比较差异均有统计学意义(P<0.05)。结论所建指纹图谱操作简便、准确,结合多元统计分析可用于评价不同产地茶芎的整体质量。阿魏酸、洋川芎内酯A、Z-藁本内酯、E-藁本内酯可能是影响不同产地茶芎药材质量的差异性成分,且前3种成分含量均以瑞昌茶芎最高,E-藁本内酯含量以德安茶芎最高。
ABSTRACT: OBJECTIVE To establish the fingerprints of Ligusticum sinense from different habitats ,screen differential components and determine their contents. METHODS Using Z-ligustilide as reference ,HPLC fingerprints of 12 batches of L. sinense were established by using Similarity Evaluation System of Chromatographic Fingerprints of TCM (2012 edition);common peaks were identified and their similarities were evaluated. Cluster analysis (CA),principal component analysis (PCA)and orthogonal partial least squares-discriminant analysis (OPLS-DA)were performed to screen differential components with variable importance in the projection (VIP)>1 as standard ;meanwhile,the contents of above differential components were determined by the same HPLC method. RESULTS There were 17 common peaks in the fingerprints of 12 batches of L. sinense ,and their similarities ranged 0.989-1.000. A total of 9 common peaks were identified ,i.e. chlorogenic acid (peak 1),ferulic acid (peak 2), senkyunolide Ⅰ(peak 7),coniferyl ferulate (peak 9),E-ligustilide(peak 13),senkyunolide A (peak 14),Z-ligustilide(peak 17). CA results showed that 12 batches of L. sinense were divided into 3 categories,S1-S5(Wuning)were clustered into one category,S6-S8(Ruichang)were clustered into one category ,S9-S12(De’an)were clustered into one category ;the VIP values of peaks 2,13,14 and 17(corresponding to ferulic acid ,E-ligustilide,senkyunolide A ,and Z-ligustilide respectively )were all greater than 1,respectively. In S 1-S5,S6-S8 and S 9-S12 samples,the contents of ferulic acid were 0.488-0.533,0.603-0.658 and 0.415-0.433 mg/g,respectively;senkyunolide A were 1.184-1.295,1.450-1.588 and 1.307-1.377 mg/g,respectively;E-ligustilide were 0.118-0.125,0.130-0.135 and 0.223-0.229 mg/g,respectively;Z-ligustilide were 7.200-7.681,8.076-8.643 and 4.508-4.996 mg/g, respectively;the differences between two groups were statisti-cally significant (P<0.05). CONCLUSIONS Established ARS-11);fingerprint is simple and accurate ,and can be used for overall quality evaluation of L. sinense from different habitats by combining with multivariate statistical analysis. Ferulic acid , senkyunolide A ,Z-ligustilide and E-ligustilide may be the differential components that affect the quality of L. sinense from different habitats ,the contents of the first 3 components in L. sinense from Ruichang are the highest ,and the content of E-ligustilide in samples from De’an is the highest.
期刊: 2022年第33卷第16期
作者: 龚伟伟,罗光明,秦倩,曾金祥,徐葱茏,刘明贵,张寿文
AUTHORS: GONG Weiwei,LUO Guangming,QIN Qian,ZENG Jinxiang,XU Conglong,LIU Minggui,ZHANG Shouwen
关键字: 茶芎;指纹图谱;多元统计分析;含量测定;质量评价;高效液相色谱法
KEYWORDS: Ligusticum sinense ;fingerprint;multivariate statistical analysis ;content determination ;quality evaluation ;HPLC
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