基于HPLC指纹图谱和化学模式识别的黄芩汤制备过程质量评价研究

Quality evaluation of the preparation process of Huangqin Decoction based on HPLC fingerprint and chemical pattern recognition

目的:分析黄芩汤制备过程中化学成分的变化,为其质量评控提供参考。方法:遵古籍张仲景《伤寒论》中黄芩汤处方,将不同产地批次的饮片混批投料,按一次煎煮(A组)、二次煎煮(B组)、冷冻干燥(C组)制成3组各12批样品,建立黄芩汤HPLC指纹图谱;计算相似度,标定特征峰;应用SPSS 24.0及SIMCA 14.1软件作HCA、PCA及PLS-DA分析,挖掘影响不同批次黄芩汤质量的标志性化合物。结果:指纹图谱标定13个共有峰。不同产地药材批次的黄芩汤图谱相似度存在差异但不显著(各批次样品相似度均大于0.9000)。指认黄芩苷、黄芩素、芍药苷、甘草苷、甘草酸5个特征峰。共有峰总面积:B组≈C组>A组。HCA、PCA分析将样品分为两类,A组为一类,B组、C组为一类。PLSDA分析找到4个潜在的质量差异标志物。综合得分评价:C组>B组>A组。说明黄芩汤煎煮2次较为合理,冷冻干燥能较好地保留其成分,10号峰、5号峰(甘草苷)、8号峰(黄芩苷)、9号峰可能是影响其质量的重要成分。结论:研究建立了黄芩汤指纹图谱,通过指纹图谱和化学模式识别较好地辨识了制备过程中影响黄芩汤质量的标志性化学成分,为其制备过程的质量评价研究奠定了理论基础。

Objective:To analyze the changes of chemical composition during the preparation of Huangqin Decoction and providing reference for its quality assessment and control.Method:According to the prescription of Huangqin Decoction in Zhang Zhongjing's "Treatise on Febrile Diseases ",12 batches of samples from different origins were mixed and fed into three groups according to primary decoction(group A),secondary decoction(group B) and freeze-drying(group C).HPLC fingerprints of Huangqin Decoction were established.The HCA,PCA and PLS-DA analyses were performed with SPSS 24.0 and SIMCA 14.1 software,in order to explore the signature compounds affecting the quality of different batches of Huangqin Decoction.Result:Thirteen common peaks were identified by fingerprinting.The similarity of Huangqin Decoction from different origins was different.However,the difference of the similarity of all batches of samples was no significant with the values greater than 0.9000.Five characteristic peaks of baicalin,baicalein,paeoniflorin,glycyrrhizin and glycyrrhetinic acid were identified.Total area of shared peaks of group B was similar with group C,and lager than that of group A.HCA and PCA analysis divided the samples into two categories,group A as one category,while group B and group C as another.PLS-DA analysis found 4 potential quality markers of quality differences.Comprehensive score evaluation:group C group B group A.The results indicateed that for Huangqin decoction,frying for 2 times was more reasonable and freeze-drying can retain its components better.Peak 10,peak 5(glycyrrhizin),peak 8(baicalin) and peak 9 may be important components affecting its quality.Conclusion:The study establish a fingerprint profile of Huangqin Decoction.The fingerprint profile and chemical pattern recognition better identify the signature chemical components affecting the quality of Huangqin Decoction in the preparation process,which lay a theoretical foundation for the quality evaluation study of its preparation process.