目的 建立4个产地21批杜仲Eucommia ulmoides雄花的高效液相色谱(HPLC)指纹图谱,为杜仲雄花的质量控制提供参考。方法 采用HPLC法,色谱柱为Agilent HC-C18色谱柱(250mm×4.6mm,5.0μm);流动相为乙腈-0.1%甲酸水溶液;梯度洗脱;体积流量0.8mL/min;柱温35℃;程序变化波长;进样量20μL。通过相似度评价、偏最小二乘法分析(partial least squares discriminant analysis,PLS-DA)、系统聚类分析(hierarchical cluster analysis,HCA)以及气候因子相关性分析等对杜仲雄花指纹图谱进行分析评价。结果 建立4个产地杜仲雄花HPLC指纹图谱,共标定15个共有峰,指认出其中6个色谱峰,分别为桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、芦丁、槲皮素;21批杜仲雄花指纹图谱与对照图谱的相似度为0.911~0.989;PLS-DA分析表明不同产地杜仲雄花样品具有一定差异,根据变量投影重要性(variable importance in the projection,VIP)筛选出7个差异性成分;差异性成分峰面积结合气候因子的相关性分析结果<显示9号峰峰面积与3、4月平均最低气温呈显著正相关,15号峰槲皮素峰面积与3月平均降水天数呈显著负相关,其余差异性成分与气候因子未呈现出明显的相关性。结论 该方法可应用于杜仲雄花的质量评价,为杜仲雄花质量控制与资源开发提供参考。

Objective To establish HPLC fingerprints of 21 batches of male flowers from Eucommia ulmoides in four producing areas and provide experimental reference for quality control and resource development of male flowers from E. ulmoides. Methods HPLC mthod was adopted. The determination was performed on a column of Agilent HC-C₁₈ (250 mm×4.6 mm, 5.0 μm); Mobile phase: 0.1% formic acid (A) and acetonitrile (B); Flow rate: 0.8 mL/min; Column temperature: 35 ℃; Injection volume: 20 μLwas used to detect organic chemicals in male flowers. The fingerprint of male flowers from E. ulmoides was evaluated via similarity evaluation, partial least squares discriminant analysis (PLS-DA), hierarchical cluster analysis and its relationship with climatic factors were also analyzed by using correlation analysis. Results HPLC fingerprints of male flowers of E. ulmoides from four producing areas were established. A total of 15 common peaks were calibrated and six chromatographic peaks were identified, which were aucubin, geniposidic acid, chlorogenic acid, geniposide, rutin and quercetin. The similarity between the established fingerprint of 21 batches and standard reference was 0.911—0.989. The results of PLS-DA analysis showed that there existed differences among male flower samples from different producing areas. Seven differential compounds were screened by variable importance in the projection (VIP). Moreover, correlation analysis between peak area of different compounds and climatic factors indicated that the peak area of No.9 was positively correlated with the average minimum temperature in March and April. The peak area of quercetin (No.15) was negatively correlated with the average precipitation days in March. The other five compounds showed no significant correlation with climate factors. Conclusion The established method in this paper could be applied to evaluate the quality of male flowers from E. ulmoides, with view to providing reference for quality control and resource development of male flowers from E. ulmoides.

R286.12

国家自然科学基金资助项目（31960323）；吉首大学校级科研项目（Jdy20080）