| 引用本文: |
袁文珺, 赵小梅, 杨志军, 俞晓英, 张立军.基于HPLC指纹图谱结合化学模式识别和多指标成分定量评价贯叶连翘药材的质量[J].湖南中医药大学学报,2025,45(7):1249-1258[点击复制] |
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| 基于HPLC指纹图谱结合化学模式识别和多指标成分定量评价贯叶连翘药材的质量 |
| 袁文珺,赵小梅,杨志军,俞晓英,张立军 |
| (兰州市中医医院 药剂科, 甘肃 兰州 730050;甘肃中医药大学, 甘肃 兰州 730000) |
| 摘要: |
| 目的 建立贯叶连翘药材HPLC指纹图谱及金丝桃苷、槲皮素、金丝桃素3种黄酮成分含量测定方法，并结合化学模式识别，评价不同产地贯叶连翘药材的质量。方法 采用HPLC-DAD技术，以Phenomenex ODS C18色谱柱（4.6 mm×250 mm,5 μm），甲醇-0.1%磷酸水溶液为流动相梯度洗脱，建立贯叶连翘药材指纹图谱，同时测定3种黄酮成分的含量；采用《中药指纹图谱相似度评价系统（2012版）》软件对12批次药材样品进行共有峰确认及相似度评价；通过SIMCA 14.1和SPSS 22.0统计分析软件，采用聚类分析（HCA）、主成分分析（PCA）和偏最小二乘法-判别分析（PLS-DA）对不同产地贯叶连翘药材进行质量评价。结果 建立了贯叶连翘药材指纹图谱，12批贯叶连翘药材的相似度均大于0.85；确定共有峰14个。通过化学模式识别分析，HCA将12批次贯叶连翘药材分为4类，反映了不同产地贯叶连翘药材的质量特征；PCA筛选出累计贡献率达到88.484%的3个主成分，得到决定贯叶连翘药材质量的5个化学成分；PLS-DA分析筛选出贯叶连翘药材中5个差异性成分，指认出3个成分（金丝桃苷、槲皮素、金丝桃素），可作为评价贯叶连翘药材质量差异的质量标志物。并对贯叶连翘药材中金丝桃苷、槲皮素及金丝桃素含量进行测定，其含量均值分别为0.461 4%、0.276 0%、0.051 4%。结论 建立的HPLC指纹图谱结合含量测定分析方法专属性强、稳定性高、分离度好，可以客观、全面、有效地用于贯叶连翘药材的质量评价，为其质量控制提供依据。 |
| 关键词: 贯叶连翘 指纹图谱 不同产地 金丝桃苷 槲皮素 金丝桃素 化学模式识别 |
| DOI：10.3969/j.issn.1674-070X.2025.07.007 |
| 投稿时间：2025-02-06 |
| 基金项目:国家自然科学基金项目（81460611）。 |
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| Quality evaluation of Hypericum perforatum L. medicinal materials based on HPLC fingerprint combined with chemical pattern recognition and multi-component quantitative analysis |
| YUAN Wenjun, ZHAO Xiaomei, YANG Zhijun, YU Xiaoying, ZHANG Lijun |
| (Pharmacy Department, Lanzhou City Hospital of Chinese Medicine, Lanzhou, Gansu 730050, China;Gansu University of Chinese Medicine, Lanzhou, Gansu 730000, China) |
| Abstract: |
| Objective To establish an HPLC fingerprint and develop a content-determining method for three flavonoid components (hyperoside, quercetin, and hypericin) in Hypericum perforatum L. medicinal materials, and to assess the quality of Hypericum perforatum L. medicinal materials from different origins by applying chemical pattern recognition. Methods HPLC-DAD technique was employed with a Phenomenex ODS C18 column (4.6 mm × 250 mm, 5 μm) and a mobile phase composed of methanol-0.1% phosphoric acid aqueous solution for gradient elution. The fingerprint of Hypericum perforatum L. medicina materials was established, and the component contents of the three flavonoids were simultaneously determined. The software Similarity Evaluation System for Traditional Chinese Medicines Fingerprint (2012 edition) was used to confirm common peaks and evaluate the similarity of 12 batches of medicinal material samples. Statistical analyses, including hierarchical cluster analysis (HCA), principal component analysis (PCA), and partial least squares-discriminant analysis (PLS-DA), were performed using SIMCA 14.1 and SPSS 22.0 software to assess the quality of Hypericum perforatum L. medicinal materials from different origins. Results The HPLC fingerprint of Hypericum perforatum L. medicinal materials was successfully established, with similarity values exceeding 0.85 for all 12 batches, and 14 common peaks were identified. Through chemical pattern recognition analysis, HCA classified the 12 batches of Hypericum perforatum L. medicinal materials into four categories, reflecting the quality characteristics of Hypericum perforatum L. medicinal materials from different origins. PCA selected three principal components with a cumulative contribution rate of 88.484%, identifying five key chemical components influencing quality. PLS-DA screened five differential components, with three (hyperoside, quercetin, and hypericin) identified as quality markers for evaluating quality variations. Furthermore, the content of hyperoside, quercetin, and hypericin were determined, with mean values of 0.4614%, 0.2760%, and 0.0514%, respectively. Conclusion The established HPLC fingerprint combined with quantitative analysis demonstrates strong specificity, high stability, and good resolution. It can be objectively, comprehensively, and effectively used for the quality evaluation of Hypericum perforatum L. medicinal materials, providing a basis for their quality control. |
| Key words: Hypericum perforatum L. fingerprint different origins hyperoside quercetin hypericin chemical pattern recognition |
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