目的 建立测定兔耳草类药材主要有效成分苯丙素苷类（毛蕊花糖苷、松果菊苷、大车前苷及鞭打绣球苷B）及环烯醚萜苷类（10-O-trans-p-methoxycinnamoyl-catalpol及10-O-[(E)-3,4-dimethoxycinnamoyl]-catalpol）的HPLC-PDA含量测定方法，并比较不同基原兔耳草的含量差异，为其质量控制方法及临床有效性提供依据。方法 采用资生堂SPOLAR C₁₈（250 mm×4.6 mm，5µm）；流动相为乙腈-0.4%甲酸溶液，梯度洗脱；体积流量1.0 mL/min；检测波长为325 nm；柱温为30 ℃；进样量为10 μL。依据测定结果使用聚类分析法和偏最小二乘法判别分析（orthogonal partial least squares discrimination analysis，OPLS-DA）法进行分析。结果 在此条件下，34批全缘兔耳草Lagotis integra、21批短筒兔耳草L. brevituba、10批圆穗兔耳草L. ramalana、6批革叶兔耳草L. alutacea、5批短穗兔耳草L. brachystachya及4批紫叶兔耳草L. praecox的6种有效成分的色谱峰分离度均良好，线性关系良好。不同基原所含的主要有效成分类型及含量均不一致，有共性也有差异性。运用SIMCA 14.1和SPSS 25.0软件分析结果显示，短筒兔耳草与圆穗兔耳草聚为一类，紫叶兔耳草与短穗兔耳草聚为一类，内在组成及含量差异小；全缘兔耳草不同产区差异大，革叶兔耳草组间差异最大。结论 建立的含量测定分析方法能有效地对多种基原兔耳草进行分析，能有效地对其进行质量控制；通过比较不同基原的兔耳草，为兔耳草资源的开发及利用提供有效依据。

Objective HPLC-PDA content determination method was established to determinate the main active ingredients of Honglian (Lagotidis Herba), which included phenylpropanoid glycosides (verproside, echinacoside, plantamajoside, hemiphroside B) and iridoid glycosides (10-O-trans-p-methoxycinnamoyl-catalpol and 10-O-[(E)-3,4-dimethoxycinnamoyl]-catalpol and compared with the other primitive species, so as to provide evidence for quality control methods and clinical effectiveness. Methods The HPLC analysis was performed on SPOLAR C₁₈ (250 mm×4.6 mm, 5µm), using acetonitrile and 0.4% formic acid solution as the mobile phase at a flow rate of 1.0 mL/min, the detection wave-length was 325 nm and column temperature was 30℃ with 10 μL injection volume. The combined determination results were analyzed using partial least squares discriminant analysis (OPLS-DA) and cluster analysis. Results Under this condition, the chromatographic peak resolution of the six active ingredients had good separationin in 34 batches of Lagotis integra,21 batches of L. brevituba, 10 batches of L. ramalana, six batches of L. alutacea, five batches of L. brachystachya Maxim.and four batches of L. praecox. The six ingredients had a good linear relationship.The types and contents of the main active ingredients contained in different bases were all inconsistent, with commonalities and differences. The results analyzed by SPSS 25.0 and SIMCA 14.1 software showed that L. brevituba and L. ramalana were clustered, L. brachystachya and L. praecox were clustered together, and there were large difference in different producing areas of L. integra, and the difference between groups of L. alutacea was the greatest. Conclusion The established content determination and analysis method could effectively analyze a variety of Lagotidis Herba and could effectively control its quality. By comparing the species from different origins, it provided an effective basis for the development and utilization of its resources.

R286.2

国家食药监总局药化司专项“特色民族药材检验方法示范性研究”；国家重点研发计划子课题（2019YFC1712302）；成都中医药大学杏林学者项目（XSGG2019023）；成都中医药大学杏林学者项目（QNXZ2019024）