基于化学模式识别法的连花清瘟胶囊HPLC指纹图谱研究

    Study on HPLC Fingerprint of Lianhua Qingwen Capsule Based on Chemical Recognition Pattern Method

    • 摘要:
      目的  采用HPLC建立连花清瘟胶囊的指纹图谱,并结合化学模式识别法对其质量进行分析评价。
      方法 采用Agilent Zorbax SB-C18(250 mm×4.6 mm,5 μm)色谱柱,以0.1%磷酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱,流速为1.0 mL·min–1,柱温为30 ℃,检测波长为207 nm。利用相似度评价、系统聚类分析(hierarchical clustering analysis,HCA)、雷达图分析、主成分分析(principal component analysis,PCA)与正交偏最小二乘判别分析(orthogonal partial least-squares discrimination analysis,OPLS-DA)对13批连花清瘟胶囊的指纹图谱进行分析评价。
      结果 建立的连花清瘟胶囊的HPLC 指纹图谱共标定了40个共有峰,通过化学对照品指认了其中10个共有峰,13批连花清瘟胶囊指纹图谱的相似度均在0.947~1.000。通过HCA与OPLS-DA等分析,13批连花清瘟胶囊样品被分为3类,分类结果与生产日期呈现相关性,并筛选出导致质量差异的8个主要化学标记物。
      结论 建立的HPLC指纹图谱稳定性、重复性好,可为连花清瘟胶囊的质量评价提供参考。

       

      Abstract:
      OBJECTIVE To establish the fingerprint of Lianhua Qingwen capsule(LHQW) with the HPLC method, and to evaluate the quality by combining with chemical pattern recognition method.
      METHODS The chromatographic separation was performed on an Agilent Zorbax SB-C18(250 mm×4.6 mm, 5 μm) column with 0.1% phosphoric acid(A)-acetonitrile(B) as the mobile phase for the gradient elution. The detection wavelength was set at 207 nm. The flow rate was set at 1.0 mL·min–1 and the column temperature was 30 ℃. Similarity evaluation, hierarchical clustering analysis(HCA), radar plot analysis, principal component analysis(PCA), and orthogonal partial least-squares discrimination analysis(OPLS-DA) were used for the further assessment of 13 batches of LHQW samples.
      RESULTS The fingerprint of LHQW was established with 40 common peaks, in which 10 common peaks were identified, and the similarities of 13 batches of LHQW samples were 0.947–1.000. By applying chemical pattern recognition methods such as HCA, and OPLS-DA, 13 batches of samples were classified into three clusters, and the results of classification were correlated with the production date. And 8 major chemical markers causing quality differences were screened.
      CONCLUSION With good reproducibility and stability, this method could provide the reference for the quality evaluation of LHQW.

       

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