摘要
目的建立补骨脂炮制前后分别纳入复方二神丸的石油醚提取物的HPLC指纹图谱,初步建立炮制前后二神丸提取物的化学模式识别方法,为其内在质量的控制提供参考。方法采用HPLC法,以思普乐C18(250 mm×4.6 mm,5μm)为色谱柱,0.01%甲酸水溶液和甲醇-乙腈(1∶1)为流动相进行梯度洗脱,检测波长210 nm,柱温30℃,体积流量1 m L/min,进样量10μL,分别建立炮制前后二神丸的提取物指纹图谱,并且采用指纹图谱相似度评价,结合聚类分析和主成分分析(PCA)进行化学模式识别研究。结果分别建立了炮制前后二神丸提取物的HPLC指纹图谱,各色谱峰分离较好,标定了20批次提取物的28个共有峰,明确了其中6个成分(补骨脂素、异补骨脂素、补骨脂酚、甲基丁香酚、甲基异丁香酚、去氢二异丁香酚);10批炮制前、后提取物分别与对照图谱比较,相似度均大于0.9;PCA和聚类分析可将20批提取物很好地分为炮制前和炮制后2类,且分类结果一致,同时PCA结果提示了一些可区别炮制前后二神丸的指纹图谱特征性的色谱峰。结论该方法稳定、可靠,为炮制前后二神丸提取物的质量控制提供了方法依据,同时结合化学模式识别研究可有助于炮制前后二神丸提取物整体质量控制及质量评价。
Objective To establish the HPLC fingerprint of Ershen Pills extracted with petroleum ether, which contains Psoraleae Fructus（PF） before and after processing, and initially set up the pattern recognition technique of the extract, so as to provide reference for quality control. Methods HPLC was used, and the separation was performed at 30 ℃ on an Spolar Hplc Packed C18 column（250 mm × 4.6 mm, 5 μm）. Gradient elution was performed with the mobile phases of methanol-acetonitrile（1∶1） and water containing 0.1% formic acid. The flow rate was 1 m L/min, and sample size was 10 μL. The UV detection wavelength was set at 210 nm. Furthermore, the fingerprint was analyzed by similarity analysis, as well as pattern recognition technique, including hierarchical clustering analysis and principal component analysis. Results The fingerprints were established respectively consisting of 28 common peaks well separated. There into, six compounds were identified（psoralen, isopsoralen, methyleugenol, methylisoeugenol, dehydrodiisoeugenol, and backuchiol）. Compared with the reference spectrum, the similarity degrees were greater than 0.9. When dealing with principal component analysis, the samples were well divided into two categories, which was consistent with the principal component analysis. Meanwhile, the results of principal component analysis prompted some chromatographic peaks which could be used to distinguish between two species. Conclusion The method is stable and reliable, which can be used for quality control of Ershen Pills extracted with petroleum ether. Meanwhile, it is contributing to the entire quality control and quality evaluation when combined with chemical pattern recognition.
出处
《中草药》
CAS
CSCD
北大核心
2017年第19期3977-3985,共9页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金面上项目(81274035)
国家基础科学人才培养基金项目(J13100340-17)
作者简介
熊瑞（1991-），女，重庆人，在读博士研究生，主要从事中药学和中药炮制原理的研究。Tel：15281090514 E-mail：342296918@qq．com
通信作者胡昌江（1952-），男，四川成都人，教授，博士研究生导师，学士学位，主要从事中药炮制学的教学和科研工作。E-mail：hhccjj@hotmail．com
