目的 建立紫菀Aster tataricus的UPLC指紋圖譜及多成分含量測定方法，并結(jié)合化學(xué)計(jì)量學(xué)方法尋找不同產(chǎn)地紫菀藥材質(zhì)量差異成分，為其質(zhì)量控制研究提供技術(shù)方法和基礎(chǔ)數(shù)據(jù)。方法 采用菲羅門Titank C₁₈色譜柱（150 mm×2.1 mm，1.8μm），以0.2%甲酸水（A）-乙腈（B）溶液為流動相，梯度洗脫（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），體積流量為0.3 mL/min，柱溫為40℃，進(jìn)樣量為2 μL，檢測波長采用定時(shí)波長（0～50 min，325 nm；50～70 min，260 nm；70～76 min，203 nm）。采用中藥指紋圖譜相似度評價(jià)軟件進(jìn)行相似度評價(jià)，結(jié)合化學(xué)計(jì)量學(xué)分析，同時(shí)對綠原酸、咖啡酸、阿魏酸、槲皮素、山柰酚、紫菀酮的含量進(jìn)行測定。結(jié)果 建立了紫菀藥材的指紋圖譜，共標(biāo)記了17個(gè)共有峰，基于對照品比對法及質(zhì)譜鑒定指認(rèn)9個(gè)色譜峰，分別為峰1（綠原酸）、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）。聚類分析（cluster analysis，CA‌）將18批紫菀樣品分為河北產(chǎn)地和安徽產(chǎn)地。主成分分析（principal component analysis，PCA）表明，不同產(chǎn)地的紫菀樣品間存在差異。通過正交偏最小二乘法-判別分析（orthogonal partial least squares-discriminant analysis，OPLS-DA）項(xiàng)下的VIP分析篩選出峰11（asterin）、12（山柰酚）、1（綠原酸）、14、10（槲皮素）、9、13（甘草素）、17（紫菀酮）可能是影響不同產(chǎn)地、不同加工方式紫菀質(zhì)量差異的標(biāo)志物。綠原酸、咖啡酸、阿魏酸、槲皮素、山柰酚、紫菀酮的質(zhì)量分?jǐn)?shù)分別為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，經(jīng)方法學(xué)考察，各成分呈現(xiàn)良好的線性關(guān)系。含量測定結(jié)果表明，熏硫?qū)ψ陷彝坑绊戄^大，經(jīng)熏硫后的紫苑中紫菀酮含量顯著降低。結(jié)論 以多指標(biāo)成分定量結(jié)合化學(xué)計(jì)量學(xué)建立的評價(jià)方法區(qū)分了不同產(chǎn)地的紫菀，評價(jià)了不同加工方式的紫菀中化學(xué)成分的含量差異，可為紫菀藥材的質(zhì)量評價(jià)與控制提供科學(xué)依據(jù)及參考。

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

河北省省級科技計(jì)劃項(xiàng)目資助（21372503D）