Department of Pharmaceutical Analysis, School of Pharmacy, Fudan University, Shanghai, P. R. China.
J Sep Sci. 2011 May;34(10):1123-32. doi: 10.1002/jssc.201000834. Epub 2011 Mar 25.
We first investigated liquid chromatography (LC) fingerprint method using multi-wavelength combination technique, and successfully used this method for the analysis of a fat-soluble extract from Radix isatidis. LC fingerprints of fat-soluble R. isatidis extracts from 11 origins were established using the Origin software and Similarity Evaluation System for Chromatographic Fingerprints of traditional Chinese medicine (TCM). The typical LC fingerprints of fat-soluble extracts from R. isatidis were first established, and the reference chromatogram was also generated with 24 common peaks showing large peak areas and good separation from adjacent peaks. Seven common characteristic peaks were identified for the first time: anthranilic acid, syringic acid, benzoic acid, salicylic acid, tryptanthrin, indigo and indirubin. The total peak areas of 24 common peaks were more than 80% of the total peak areas. Hierarchical clustering analysis (HCA) of 11 R. isatidis samples was performed, and the results show that the differences between 11 origin R. isatidis were large. Principal component analysis (PCA) on 24 common peaks was obtained to find the possible chemical markers for the discrimination of different samples. The loading plot indicated that peaks 8, 11, 13 and 14 may have more influence on the discrimination of the samples. All these were useful for evaluating and controlling the quality of R. isatidis. Our work provides a general model of chromatogram combination at multi-wavelength detection to study the complex or the undeveloped materials, which can be used to scientifically ensure the quality of such samples and deeply do qualitative, quantitative and multicomponent pharmacodynamic research combined with modern advanced chromatographic technique.
我们首先研究了使用多波长组合技术的液相色谱（LC）指纹图谱法，并成功地将该方法用于分析板蓝根脂溶性提取物。使用 Origin 软件和中药色谱指纹图谱相似度评价系统（TCM）建立了来自 11 个产地的脂溶性板蓝根提取物的 LC 指纹图谱。首先建立了脂溶性板蓝根提取物的典型 LC 指纹图谱，并生成了参考图谱，图谱上有 24 个常见峰，峰面积较大，与相邻峰分离良好。首次鉴定出 7 个共有特征峰：邻氨基苯甲酸、丁香酸、苯甲酸、水杨酸、色氨酸、靛蓝和靛玉红。24 个共有峰的总峰面积超过总峰面积的 80%。对 11 个板蓝根样品进行了层次聚类分析（HCA），结果表明 11 个产地的板蓝根差异较大。对 24 个共有峰进行主成分分析（PCA），以寻找区分不同样品的可能化学标志物。加载图表明，峰 8、11、13 和 14 可能对样品的区分有更大的影响。所有这些都有助于评价和控制板蓝根的质量。我们的工作提供了一种在多波长检测下进行色谱组合的一般模型，用于研究复杂或未开发的材料，可以结合现代先进的色谱技术，用于科学地保证此类样品的质量，并深入进行定性、定量和多成分药效学研究。
