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目的建立不同产地马兜铃蜜炙前后的HPLC指纹图谱,比较分析马兜铃蜜炙前后化学成分变化。方法采用高效液相色谱分析,Agilent ZORBAX SB-C_(18)柱(4.6 mm×250 mm,5μm),流动相为乙腈-1%乙酸溶液,梯度洗脱,流速1.0 mL·min~(-1),检测波长250 nm,柱温30℃;采用《中药色谱指纹图谱相似度评价软件》、SPSS22.0软件对HPLC色谱进行相似度分析、聚类分析及主成分分析。结果建立14批不同产地马兜铃蜜炙前后的指纹图谱,蜜炙后马兜铃酸Ⅰ(AAⅠ)降低率达36%以上;相似度评价显示生、制马兜铃相似度大于0.9(除S4为0.822,S18为0.771,S19为0.639);聚类分析表明,生品马兜铃大致分为4类,主成分分析可知,5个主成分(包括AA I)的累积方差贡献率为84.914%,从化学成分上反映了不同产地样品的相似性及差异性。结论本实验结果可以为马兜铃炮制减毒及炮制工艺的规范化提供参考,具有示范性。
OBJECTIVE To establish the HPLC fingerprints of Aristolochia bicolor before and after Sunburn in different areas and analyze the changes of chemical composition before and after the Sunburn. Methods High performance liquid chromatography (HPLC) was performed on an Agilent ZORBAX SB-C 18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase of acetonitrile-1% acetic acid and gradient elution at a flow rate of 1.0 mL · min -1 ), The detection wavelength was 250 nm, and the column temperature was 30 ℃. The similarity analysis, cluster analysis and principal component analysis of HPLC chromatograms were carried out by using the software of similarity evaluation of chromatographic fingerprints of Chinese traditional medicine. Results Fingerprints of 14 batches of Aristolochia nipponense were obtained before and after the treatment. The reduction rate of AA Ⅰ after honey-burning was above 36%. The similarity evaluation showed that the similarity of raw and prepared aristolochia was greater than 0.9 S4 was 0.822, S18 was 0.771 and S19 was 0.639). The cluster analysis showed that the crude product was divided into four categories. The principal components analysis showed that the cumulative variance of the five principal components (including AA I) was 84.914 %, From the chemical composition reflects the similarities and differences in samples of different origin. Conclusion The results of this experiment can provide a reference for the standardization of processing Aristolochia concanavalin processing and processing, with a demonstration.