北刘寄奴标准汤剂指纹图谱及含量测定研究

Fingerprint and Content Determination of Siphonostegia chinensis Benth.Standard Decoction

目的:建立北刘寄奴标准汤剂的高效液相色谱法(HPLC)指纹图谱和多成分定量分析方法。方法:采用HPLC建立北刘寄奴标准汤剂指纹图谱,并采用“中药色谱指纹图谱相似度评价系统”(2012版)进行相似度评价。通过聚类分析(CA)、主成分分析(PCA)及正交偏最小二乘法-判别分析(OPLS-DA),以变量重要性投影(VIP)值>1为标准筛选影响质量差异的标志物。采用HPLC测定木犀草素-7-O-β-D-葡萄糖醛酸苷、毛蕊花糖苷、木犀草素的含量。结果:北刘寄奴标准汤剂指纹图谱有11个共有峰,各批样品相似度均大于0.90,指认峰1为咖啡酸、峰3为木犀草素-7-O-β-D-葡萄糖醛酸苷、峰4为毛蕊花糖苷、峰6为芹菜素-7-O-β-D-葡萄糖醛酸苷、峰7为异类叶升麻苷、峰10为木犀草素、峰11为芹菜素。CA结果表明,18批样品分为4类:S16~S18为一类、S4和S12为一类,S9、S11和S14为一类,其余批样品聚为一类。PCA结果表明,3个主成分的累积方差贡献率为89.019%。OPLS-DA表明,峰11、峰9、峰7、峰5、峰4、峰10是影响北刘寄奴标准汤剂质量的关键性成分。18批北刘寄奴标准汤剂中毛蕊花糖苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、木犀草素质量分数分别为0.38%~2.17%、0.13%~0.73%、0.06%~0.21%。结论:建立的指纹图谱和含量测定方法操作简单,具有良好的重复性和重现性,可用于评价北刘寄奴标准汤剂的质量差异,对其质量控制及整体性评价具有一定的意义。

Objective:To establish a high-performance liquid chromatography(HPLC)fingerprint and multi-component quantitative analysis method for Siphonostegia chinensis Benth.standard decoction.Methods:The HPLC method was used to establish the fingerprint of S.chinensis Benth.standard decoction,and the similarity was evaluated using the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2012 version).Cluster analysis(CA),principal component analysis(PCA),and orthogonal partial least squares-discriminant analysis(OPLS-DA)were performed to screen marker compounds influencing quality differences based on a variable importance projection(VIP)value>1.The content of luteolin-7-O-β-D-glucuronide,verbascoside,and luteolin was determined by HPLC.Results:The fingerprint of S.chinensis Benth.standard decoction showed 11 common peaks,with a similarity greater than 0.90 among all samples.Peak 1 was identified as caffeic acid,peak 3 as luteolin-7-O-β-D-glucuronide,peak 4 as verbascoside,peak 6 as apigenin-7-O-β-D-glucuronide,peak 7 as isoacteoside,peak 10 as luteolin,and peak 11 as apigenin.Cluster analysis divided the 18 batches of samples into four categories:S16-S18,S4 and S12,S9,S11,and S14,and the remaining batches clustered into one group.Principal component analysis showed that the cumulative variance contribution rate of the three principal components was 89.019%.OPLS-DA indicated that peaks 11,9,7,5,4,and 10 were key components affecting the quality of S.chinensis Benth.standard decoction.The content of verbascoside,luteolin-7-O-β-D-glucuronide,and luteolin in the 18 batches of S.chinensis Benth.standard decoction ranged from 0.38%to 2.17%,0.13%to 0.73%,and 0.06%to 0.21%,respectively.Conclusion:The established fingerprint and quantitative analysis methods are simple to operate and have good repeatability and reproducibility.They can be used to evaluate the quality differences of the standard decoction of S.chinensis Benth.and have certain significance for its quality control and holistic evaluation.