目的:建立能有效区分葛花(Pueraria lobata (Willd.) Ohwi)、粉葛花(Pueraria thomsonii Benth.)及其混淆品种的特征图谱和多指标成分测定分析方法,用于葛花药材的整体质量控制。方法:采用高效液相色谱法(HPLC)对不同商品规格葛花中主要化学成分进行分析,确定最佳色谱条件,建立特征图谱;利用主成分分析(PCA)和偏最小二乘判别分析(PLS-DA)对不同商品规格葛花特征图谱数据进行分析,筛选确认差异性标志物;通过与对照品比对,对差异性标志物进行指认,进行定量分析,并分析测定结果。结果:建立了葛花、粉葛花、葛麻姆花及紫藤花的特征图谱,分别匹配了18、27、18及8个共有峰,指认出葛根素(Pu)、大豆苷(Da)、大豆苷元(Dae)、染料木苷(Ge)、染料木素(Gee)、葛花苷(Ka)、glycitin(Gl)、tectoridin(Td)、tectorigenin(Tg)及tectorigenin-7-O-xylosylglucoside(Tx)10个色谱峰,并同时建立了上述10个成分的含量测定方法。PCA和PLS-DA结果可将4种不同商品规格葛花样品明显区分,以Gl、Tx、Ge、Td、Ka及未知化合物(6号峰)为差异性标志物。葛花、粉葛花所含成分明显区别其混淆品,葛花中Ka含量为粉葛花的10倍,Ge含量为粉葛花的50%,未检测到Da、Gl。结论:所建立的特征图谱结合化学计量学及多指标成分测定分析方法简便、可行,可客观、有效地用于葛花药材质量控制,作为修改葛花质量标准提供参考。
张立军, 李鹏, 李隆海, 张新玥, 马冬妮, 张转平, 胡芳弟, 王磊, 柯红梅, 李鹏, 刘丽, 吴春燕, 李小安
. 基于特征图谱、化学计量学和多指标成分测定的不同商品规格葛花质量差异分析*[J]. 药物分析杂志, 2024
, 44(5)
: 903
-911
.
DOI: 10.16155/j.0254-1793.2024.05.19
Objective: To establish HPLC characteristic chromatogram and multi-index component determination methods, so as to effectively distinguish Pueraria lobata (Willd.) Ohwi (GH), Pueraria thomsonii Benth (FGH) and their confusing species, and to evaluate the quality of Puerariae Flos. Methods: HPLC was used to analyze the chemical components from different Puerariae Flos with different commercial specifications. The chromatographic conditions were optimized, and the characteristic chromatogram was established. Principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) analysis were used to analyze the characteristic chromatogram data of Puerariae Flos with different commercial specifications and to select the differential markers of Puerariae Flos. The differential markers were identified by comparing them with the reference substance, and their contents were determined and the determination results were analyzed. Results: HPLC characteristic chromatograms of GH, FGH and their confusing species were established, and a total of 18, 27, 18 and 8 common peaks were matched in GH, FGH, Pueraria lobata var. montana (GMM) and Wisteria sinensis (Sims) Sweet (ZTH) respectively. Ten chromatographic peaks were identified, including puerarin (Pu), daidzin (Da), daidzein (Dae), genistin (Ge), genistein (Gee), kakkalide (Ka), glycitin (Gl), tectoridin (Td), tectorigenin (Tg) and tectorigenin-7-O-xylosylglucoside (Tx), and HPLC method for the determination of the above ten components was established simultaneously. The results of PCA and PLS-DA could distinguish four kinds of samples with different commercial specifications. Gl, Tx, Ge, Td, Ka and one unknown compound 6 were the components with significant differences between the samples with four different commodity specifications. The chemical components of GH and FGH were obviously different from their adulterants. The content of Ka in GH were 10 times as much as that in FGH, and the content of Ge was half as much as that in FGH. Da, Gl was not detected in GH. Conclusion: The established characteristic chromatogram, chemometrics and multi-index component determination method in this study is rapid, simple, objectively and effectively and comprehensive quality evaluation of Puerariae Flos, which can provide the references for improving the quality control standard of Puerariae Flos.
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