经典名方泽泻汤的HPLC指纹图谱及多指标含量测定研究

HPLC fingerprint and multi-components determination of classical herbal prescription Zexie Decoction

目的建立经典名方泽泻汤的HPLC指纹图谱,结合化学模式识别技术对其进行分析,并测定泽泻汤中3种成分23-乙酰泽泻醇B、23-乙酰泽泻醇C和白术内酯Ⅲ的含量,为泽泻汤质量控制提供科学依据。方法采用Waters WAT054275C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-水为流动相进行梯度洗脱,建立15批泽泻汤的HPLC指纹图谱,并采用"中药色谱指纹图谱相似度评价系统"、SPSS22.0及SIMCA14.1软件对15批泽泻汤的HPLC指纹图谱进行相似度评价及聚类分析(CA)、偏最小二乘判别分析(PLS-DA)等化学模式识别。同时测定23-乙酰泽泻醇B、23-乙酰泽泻醇C和白术内酯Ⅲ的含量。结果建立了15批泽泻汤的HPLC指纹图谱,相似度均>0.94,标定了18个共有峰,指认出3个色谱峰,分别为11号峰白术内酯Ⅲ、15号峰23-乙酰泽泻醇C、16号峰23-乙酰泽泻醇B,CA和PLS-DA将15批泽泻汤样品分为2类,同时15批泽泻汤中23-乙酰泽泻醇B、23-乙酰泽泻醇C和白术内酯Ⅲ的质量分数分别为0.321~0.569、0.075~0.139、0.106~0.142 mg/g。结论 HPLC指纹图谱结合多成分同时测定的方法,快速、简便、重复性好,为泽泻汤及其制剂的质量评价提供参考。

Objective To establish fingerprints of Zexie Decoction(ZXD) by HPLC, analyze them with chemical pattern recognition technology, and determine the contents of 23-acetyl alisone B, 23-acetyl alisone C, and atractylode Ⅲ in ZXD, in order to provide a scientific basis for its quality control. Methods The HPLC fingerprint of ZXD was performed on Waters WAT054275 C18 column(250 mm × 4.6 mm, 5 μm) with the mobile phase consisting of acetonitrile and water. Similarity Evaluation System for Chromatographic Fingerprint of TCM(2004 A edition) was used to evaluate the fingerprints. SPSS 22.0 software and SIMCA 14.1 software was used for cluster analysis and discriminant analysis of partial least squares of those samples. Furthermore, the content of 23-acetyl alisone B, 23-acetyl alisone C, and atractylode Ⅲ was determined. Results The HPLC fingerprint of 15 batches of ZXD was established. The similarity was > 0.94, and 18 common peaks were marked. Three peaks were confirmed, they were: atractylode Ⅲ(peak 11), 23-acetyl alisone C(peak 15), and 23-acetyl alisone B(peak 16) were confirmed. Cluster analysis and partial least square discriminant analysis were used to classify the 15 batches of ZXD samples into two groups. The mass fraction of 23-acetyl alisone B, 23-acetyl alisone C, and atractylode Ⅲ in 15 batches of ZXD were in the range of 0.321-0.569, 0.075-0.139, and 0.106-0.142 mg/g, respectively. Conclusion The combination methods of HPLC fingerprint and simultaneous determinations of multiple components are rapid, simple and reproducible, which can provide reference for the quality evaluation of ZXD and its preparation.