基于化学模式识别方法分析止得咳颗粒的抗炎谱效关系

Analysis on Spectrum-effect Relationship of Anti-inflammatory Effect of Zhideke Granules Based on Chemical Pattern Recognition

目的:研究止得咳颗粒HPLC指纹图谱与抗炎作用的谱效关系,探索其抗炎药效物质基础。方法:采用HPLC建立10批止得咳颗粒的指纹图谱并进行色谱峰归属,流动相甲醇-0.2%磷酸水溶液梯度洗脱,检测波长254 nm。综合应用相似度评价,聚类分析(HCA),主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)评价止得咳颗粒批间质量差异。采用偏最小二乘法(PLS)将指纹图谱共有峰与二甲苯致小鼠耳肿胀和小鼠棉球肉芽肿的抑制率进行关联性分析,筛选出对抗炎药效作用贡献较大的色谱峰。结果:止得咳颗粒HPLC指纹图谱标定了31个共有峰,10批样品相似度均≥0.992。HCA与PCA分析结果一致,均将样品分为3类;OPLS-DA结果发现15个成分是影响样品批间差异的主要标记物。不同批次止得咳颗粒样品均有不同程度的抗炎作用,PLS分析得出与抗炎作用呈正相关的色谱峰主要来自射干、黄芩,贡献较大的色谱峰为3,6,19,27~30号峰,其中28,30号峰依次为次野鸢尾黄素、汉黄芩素。结论:HPLC指纹图谱结合化学模式识别方法可为系统评价止得咳颗粒整体质量提供参考。止得咳颗粒对小鼠急、慢性炎症反应有一定的抑制作用,抗炎作用是多种成分共同作用的结果,射干、黄芩对该制剂发挥抗炎作用具有显著意义。

Objective:The aim of this study was to research the relationship between HPLC fingerprint and anti-inflammatory effect of Zhideke granules,and the substance basis of its anti-inflammatory effect was preliminary explored.Method:The fingerprint of 10 batches of Zhideke granules were determined by HPLC,the mobile phase was consisted of methanol-0.2%phosphoric acid solution for gradient elution with a detection wavelength of 254 nm.Similarity analysis,hierarchical cluster analysis(HCA),principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were used to evaluate the quality difference between batches of Zhideke granules.The correlation analysis between the common peaks and the inhibition rates of Zhideke granules on ear swelling and cotton ball granuloma in mice was carried out by partial least squares(PLS),and the peaks greatly contributing to the anti-inflammatory effect were screened out.Result:There were 31 common peaks in the HPLC fingerprint of Zhideke granules.The similarities of 10 batches samples were≥0.992.The HCA and PCA analysis results were consistent,and the samples were divided into 3 categories.Combined with the OPLS-DA results,15 components were the main markers affecting the differences of different batches of samples.Different batches of Zhideke granules differed in anti-inflammatory effect.The chromatographic peaks being positively correlated with the anti-inflammatory effect were mainly from Belamcandae Rhizoma and Scutellariae Radix,Chromatographic peaks 3,6,19,27-30 had significant contribution to anti-inflammatory effect,of which peaks 28 and 30 were irisflorentin and wogonin.Conclusion:HPLC fingerprint combined with chemical pattern recognition method can provide a reference for systematic evaluation of the overall quality of Zhideke granules.Zhideke granules has a certain inhibitory effect on acute and chronic inflammation in mice,and the anti-inflammatory effect is the result of the combined action of various ingredients,while Belamcandae Rhizoma and Scutellariae Radix have significant significance for the antiinflammatory effect.