目的 建立紫菀Aster tataricus的UPLC指纹图谱及多成分含量测定方法，并结合化学计量学方法寻找不同产地紫菀药材质量差异成分，为其质量控制研究提供技术方法和基础数据。方法 采用菲罗门Titank C₁₈色谱柱（150 mm×2.1 mm，1.8μm），以0.2%甲酸水（A）-乙腈（B）溶液为流动相，梯度洗脱（0～3 min，5%～9% B；3～13 min，9%～10% B；13～15 min，10%～16% B；15～20 min，16% B；20～50 min，16%～28% B；50～54 min，28%～51% B；54～64 min，51%～100% B；64～74 min，100% B；74～76 min，100%～5% B），体积流量为0.3 mL/min，柱温为40℃，进样量为2 μL，检测波长采用定时波长（0～50 min，325 nm；50～70 min，260 nm；70～76 min，203 nm）。采用中药指纹图谱相似度评价软件进行相似度评价，结合化学计量学分析，同时对绿原酸、咖啡酸、阿魏酸、槲皮素、山柰酚、紫菀酮的含量进行测定。结果 建立了紫菀药材的指纹图谱，共标记了17个共有峰，基于对照品比对法及质谱鉴定指认9个色谱峰，分别为峰1（绿原酸）、峰3（咖啡酸）、峰5（阿魏酸）、峰10（槲皮素）、峰11（asterin）、峰12（山柰酚）、峰13（甘草素）、峰16（豆甾醇葡萄糖苷）、峰17（紫菀酮）。聚类分析（cluster analysis，CA‌）将18批紫菀样品分为河北产地和安徽产地。主成分分析（principal component analysis，PCA）表明，不同产地的紫菀样品间存在差异。通过正交偏最小二乘法-判别分析（orthogonal partial least squares-discriminant analysis，OPLS-DA）项下的VIP分析筛选出峰11（asterin）、12（山柰酚）、1（绿原酸）、14、10（槲皮素）、9、13（甘草素）、17（紫菀酮）可能是影响不同产地、不同加工方式紫菀质量差异的标志物。绿原酸、咖啡酸、阿魏酸、槲皮素、山柰酚、紫菀酮的质量分数分别为5.941 6～18.745 1、0.477 2～1.046 6、0.177 4～0.265 6、1.135 8～1.720 7、0.574 9～2.755 7、148.340 8～252.163 9 μg/g，经方法学考察，各成分呈现良好的线性关系。含量测定结果表明，熏硫对紫菀酮含量影响较大，经熏硫后的紫苑中紫菀酮含量显著降低。结论 以多指标成分定量结合化学计量学建立的评价方法区分了不同产地的紫菀，评价了不同加工方式的紫菀中化学成分的含量差异，可为紫菀药材的质量评价与控制提供科学依据及参考。

Objective To establish an ultra-high performance liquid chromatography (UPLC) fingerprint and multi-component content determination method for Ziwan (Asteris Radix et Rhizoma) from different sources, and evaluate the quality of Asteris Radix et Rhizoma from different sources in combination with chemometrics methods, so as to provide technical method reference and basic data for its quality control research. Methods Phenomenex Titank C₁₈ chromatographic column (150 mm×2.1 mm, 1.8 μm ) was used, with 0.2% formic acid water (A) - acetonitrile (B) solution as mobile phase, gradient elution (0—3 min, 5%—9% B; 3—13 min, 9%—10% B; 13—15 min, 10%—16% B; 15—20 min, 16% B; 20—50 min, 16%—28% B; 50—54 min,28%—51% B; 54—64 min, 51%—100% B; 64—74 min,100% B;74—76 min, 100%—5% B). The flow rate was 0.3 mL/min, the column temperature was 40 ℃, the injection volume was 3 μL, and the detection wavelength was set at a fixed wavelength (0—50 min, 325 nm; 50—70 min, 260 nm; 70—76 min, 203 nm). The fingerprints of Asteris Radix et Rhizoma from different sources were constructed, and the contents of chlorogenic acid, caffeic acid, ferulic acid, quercetin, kaempferol and shionone were determined by similarity analysis and chemometrics analysis. ResultsThe fingerprints of Asteris Radix et Rhizoma were established. A total of 17 common peaks were marked. Nine chromatographic peaks were identified by the comparison method of reference substances and MS identification, which were peak 1 (chlorogenic acid), peak 3 (caffeic acid), peak 5 (ferulic acid), peak 10 (quercetin), peak 11 (asterin), peak 12 (kaempferol), peak 13 (liquiritigenin), peak 16 (stigmasterol glucoside), and peak 17 (shionone). Eighteen batches of Asteris Radix et Rhizoma samples were divided into Hebei origin and Anhui origin by cluster analysis (CA). Principal component analysis (PCA) showed that there were differences among Asteris Radix et Rhizoma samples from different habitats. According to the VIP analysis under orthogonal partial least squares-discriminant analysis (OPLS-DA), peaks 11 (asterin), 12 (kaempferol), 1 (chlorogenic acid), 14, 10 (quercetin), 9, 13 (liquiritigenin), 17 (shionone) may be a marker affecting the quality of Asteris Radix et Rhizoma from different habitats and different processing methods. The contents of chlorogenic acid, caffeic acid, ferulic acid, kaempferol, quercetin and shionone ranged from 5.941 6—18.745 1, 0.477 2—1.046 6,0.177 4—0.265 6,1.135 8—1.720 7, 0.574 9—2.755 7, 148.3 408—252.1 639 μg/g. According to the methodology, each component showed a good linear relationship. The content determination results showed that sulfur fumigation of Asteris Radix et Rhizoma had a great effect on the content of shionone, and the content of shionone decreased significantly. Conclusion The quality of Asteris Radix et Rhizoma from different habitats was distinguished by the evaluation method established by quantitative analysis of multi index components combined with chemometrics, and the content differences of chemical components in Asteris Radix et Rhizoma from different processing methods were evaluated, which could provide scientific basis and reference for the quality evaluation and control of Asteris Radix et Rhizoma.

R286.2

河北省省级科技计划项目资助（21372503D）