不同产地生/醋炙乌药指纹图谱的建立及模式识别研究

目的：采用HPLC法建立不同产地生/醋炙乌药的指纹图谱，并结合相似度计算和化学模式识别方法，评价生/醋炙乌药质量。方法：采用Agilent 5 HC-C₁₈（2）（250 mm×4.6 mm，5 μm）色谱柱，以甲醇-0.05%甲酸水溶液为流动相，梯度洗脱，流速1 mL·min^-1，检测波长235 nm，柱温（25±5）℃，进样量10 μL。采用中药指纹图谱相似度软件对53批生/醋炙乌药进行测定，计算相似度。再将数据导入SIMCA 13.0软件并进行主成分分析（principal component analysis，PCA）和正交最小二乘法-判别分析（orthogonal partial least squares discriminate analysis，OPLS-DA）。结果：建立了53批生/醋炙乌药HPLC指纹图谱；中药指纹图谱相似度软件评价生/醋炙乌药之间相似度，可得生/醋炙乌药之间相似度均在0.51~0.70，表明生/醋炙乌药差异性较大；PCA可将生/醋炙乌药基本上分为两类；OPLS-DA筛选差异标记物，确定差异贡献最大的3个成分为去甲异波尔定、乌药内酯、乌药醚内酯。结论：该方法为生/醋炙乌药质量控制提供了方法依据，同时结合化学模式识别研究有助于乌药炮制前后整体质量控制及质量评价。

Objective:To establish the fingerprint of crude and vinegar processed Linderae Radix from different habitats with HPLC method, and to evaluate the quality through the similarity calculation and chemical pattern recognition. Methods:The separation was performed on an Agilent 5 HC-C₁₈(2) (250 mm×4.6 mm, 5 μm) at(25±5)℃. A binary gradient elution system was composed of methanol and water containing 0.5% formic acid and the flow rate was 1.0 mL·min^-1. The detection wavelength was 235 nm. Similarities of fingerprints of 53 batches of crude and vinegar processed Linderae Radix were analyzed by software "Similarity Evaluation System for Chromatographic Fingerprint of TCMs"(Version 2012A). Then principal component analysis(PCA) and orthogonal partial least squares discriminate analysis(OPLS-DA) were performed with SIMCA 13.0. Results:The HPLC fingerprints of 53 batches of crude and vinegar processed Linderae Radix were established successfully in this study. The similarities of crude and vinegar processed Linderae Radix were between 0.51 and 0.70, indicating the difference between the crude and processed samples. All the samples were classified into two categories by PCA. The OPLS-DA result showed that the primary differentiating components were norisoboldine, linderalactone and linderane. Conclusion:The proposed method provided basis for quality control of crude and vinegar processed Linderae Radix. And chemical pattern recognition is proved to be helpful in comprehensive quality control and evaluation of Linderae Radix before and after the process.