目的 建立道地产区及不同主产区金银花HPLC指纹图谱，结合化学计量学评价为金银花质评质控体系的构建及其潜在质量标志物的探索提供参考和依据。方法 采集河南、山东、河北产区的金银花样品，优化提取与检测方法，建立最佳HPLC指纹图谱条件。色谱柱：Symmetry C₁₈（250 mm×4.6 mm，5μm）；流动相为乙腈（A）-0.2%甲酸水溶液（B）；梯度洗脱条件，进样体积10 μL；检测波长245 nm；体积流量0.5 mL/min；柱温38℃。采用中药色谱指纹图谱相似度评价系统（2012.130723）对HPLC图谱数据进行共有峰确定、相似度评价。采用SIMCA-P 14.1软件进行聚类分析（cluster analysis，HCA）、主成分分析（principal component analysis，PCA）、正交偏最小二乘判别分析（orthogonal partial least squares discriminant analysis，OPLS-DA）。结果 3个产地金银花HPLC指纹图谱共匹配45个共有峰成分，标示出16个色谱峰。HCA与PCA结果可将河南产区与山东、河北产区金银花样品完全区分。OPLS-DA分析结果表明异绿原酸B、香豆酸、莫诺苷、芦丁、新绿原酸、断氧化马钱苷等物质含量在河南产金银花样品中普遍高于山东、河北产金银花；绿原酸、异绿原酸C、木犀草苷在山东产金银花样品中普遍高于河南、河北产金银花；忍冬苷在河北产金银花样品中普遍高于河南、山东产金银花。结论 所建立的金银花HPLC指纹图谱结合化学计量学的方法，可应用于金银花质量标准的研究与开发及其潜在质量标志物和道地因子的探索。

Objective To establish HPLC fingerprints of Lonicera Japonica Flos from different producing areas, in order to provide reference and basis for the construction of quality evaluation system and the exploration of potential quality markers of Lonicera Japonica Flos combined with chemometrics. Methods The extraction and detection methods of Lonicera Japonica Flos from Henan, Shandong and Hebei were optimized, and the optimal HPLC fingerprint conditions were established. Chromatographic column:symmetry C₁₈(250 mm×4.6 mm, 5 μm); mobile phase:acetonitrile (a)-0.2% formic acid aqueous solution (b); gradient elution conditions; injection volume:10 μL; detection wavelength:245 nm; flow rate:0.5%. The column temperature was 38 ^oC. HPLC fingerprint similarity evaluation system (2012.130723) was used to determine common peaks and evaluate similarity. Cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis were performed by SIMCA-P 14.1 software. Results The HPLC fingerprints of Lonicera Japonica Flos from three regions matched 45 common peaks and 16 peaks were identified. The results of HCA and PCA can completely distinguish the samples from Shandong and Hebei provinces. The results of OPLS-DA showed that the samples from Henan Province contained the most isochlorogenic acid B, cumalic acid, morroniside; troxerutin, neochlorogenic acid and secoxyloganin; The samples from Shandong Province contained the most chlorogenic acid, isochlorogenic acid C and cynaroside; the samples from Hebei Province contained the most xylostein. Conclusion The HPLC fingerprint of Lonicera Japonica Flos combined with chemometrics can be applied to the research and development of the quality standard of Lonicera Japonica Flos and the exploration of its potential Q-Markers and daodi factors.

R286.2

国家自然科学基金资助项目(81603232);河南中医药大学博士科研基金资助项目(BSJJ2015-13)