| 摘要: |
| 目的 建立白花蛇舌草Hedyotis diffusa Willd HPLC指纹图谱,并对不同产地、采收期、野生与栽培白花蛇舌草及混淆品进行对比分析。方法 采用Kromasil C-18色谱柱(4.6×250mm,5μm),流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,流速为1.0mL.min-1,波长为330nm,柱温为 30 ℃,进样量为10μL。结合相似度评价、聚类分析(cluster analysis,CA)、主成分分析(principal component analysis,PCA)、正交偏最小二乘法-判别式分析(orthogonal partial least squares method discriminant analysis,OPLS-DA)比较不同产地、采收期、野生与栽培白花蛇舌草及其混淆品的差异。结果 HPLC指纹图谱共标定了白花蛇舌草12个共有峰,并指认了水晶兰苷、隐绿原酸、白麻苷、槲皮素-3-O-桑布双糖苷、芦丁、槲皮素-3-O-[2-O-(6-O-E-芥子酰基)-β-D-吡喃葡萄糖基]-β-D-吡喃半乳糖苷、槲皮素-3-O-[2-O-(6-O-E-阿魏酰基)-β-D-吡喃葡糖基]-β-D-吡喃半乳糖苷、(E)-6-O-对香豆酰鸡屎藤次苷甲酯、山奈酚-3-O-[2-O-(6-O-E-阿魏酰)-β-D-葡萄糖基]-β-D-半乳糖苷、(E)-6-O-阿魏酰鸡屎藤次苷甲酯、(E)-6-O-对香豆酰鸡屎藤次苷甲酯异构体。通过相似度分析发现,白花蛇舌草的不同产地和采收期、野生与栽培样品均存在一定的差异,与混淆品样品有明显差异；通过CA、PCA发现,30批白花蛇舌草样品存在产地交叉的现象,说明产地不是影响其质量的关键因素；通过CA、PCA、OPLS-DA 可将40批样品根据不同采收期、野生与栽培、基原进行区分,通过 OPLS-DA 项下的投影中的变量重要性投影(variable importance in projection,VIP)各筛选出6个差异性标志物。结论 建立的指纹图谱方法能准确有效区分白花蛇舌草不同采收期、野生与栽培及其混淆品,为白花蛇舌草的质量评价及鉴别提供参考。 |
| 关键词: 白花蛇舌草 HPLC指纹图谱 化学模式识别 相似度评价 质量评价 |
| DOI： |
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| 基金项目:2024年度国家药品标准制修订研究课题（2024Z001） |
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| Quantitative evaluation of Hedyotis diffusa Willd quality based on HPLC fingerprint combined with chemical pattern recognition |
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Huang Lan1, Chen Bilian2
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1.浙江中医药大学;2.Zhejiang Institute for Food and Drug Control
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| Abstract: |
| ABSTRACT: OBJECTIVE To establish HPLC fingerprint for Hedyotis diffusa Willd and analyze them with different regions, harvesting periods, wild and cultivated and adulterants. METHODS The HPLC analysis was performed on a Kromasil C-18 column (4.6×250mm，5μm) by using a mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution in a gradient elution at a flow rate of 1.0 mL·min-1, with a detection wavelength of 330 nm, and a column temperature of 30 ℃ and the injection volume of 10 μL. Similarity evaluation, cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares method discriminant analysis (OPLS-DA)
were utilized to compare the differences among different regions and harvesting periods and wild and cultivated of
Hedyotis diffusa Willd and its adulterants.RESULTS The HPLC fingerprints identified 12 common peaks for Hedyotis diffusa Willd, including eleven compounds: Monotropein、Cryptochlorogenic acid、Baimaside、Quercetin-3-O-sambubioside、Rutin、Quercetin-3-O-[2-O-(6-O-E-sinapoyl)-β-D-glucopyranosyl-β-D-galactopyranoside、Quercetin-3-O-[2-O-(6-O-E-feruloyl)-β-D-glucopyranosyl]-β-D-galactopyranoside、 (E)-6-O-Coumaroylscandoside methyl ester、Kaempferol 3-O-(2-O-(6-O-E-feruloyl)-β-D-glucopyranosyl]-β-D-galactopyranoside、(E)-6-O-Feruloylscandoside methyl ester、(E)-6-O-Feruloylscandoside methyl ester isomer. Similarity analysis found that there were some differences between the wild and cultivated samples in different
places and harvesting periods, and there were obvious differences between the samples and the confused ones. But CA and PCA found that 30 batches of samples had cross-origin phenomenon, indicating that origin was not the key factor affecting its quality. CA，PCA，and OPLS-DA could differentiate 40 batches of samples according to different harvesting periods ，wild and cultivated,and botanical origins, and OPLS-DA identified six differential markers through the variable importance in projection (VIP). CONCLUSION The established fingerprint method can accurately and effectively differentiate between different harvesting periods and wild and cultivated Hedyotis diffusa Willd, as well as its adulterants, which provides reference for quality evaluation and identification of Hedyotis diffusa Willd. |
| Key words: hedyotis diffusa Willd HPLC fingerprints chemical pattern recognition similarity evaluation quality evaluation |
