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    高效液相色谱指纹图谱结合化学计量学评价桔梗汤物质基准的质量

    Quality Evaluation of Jiegeng Decoction Material Benchmarks by High Performance Liquid Chromatography Fingerprint with Chemometrics

    • 摘要: 为研究桔梗汤物质基准的质量控制方法,进行了题示研究。以12批桔梗汤物质基准为研究对象,建立高效液相色谱(HPLC)指纹图谱,与对照指纹图谱对比,指认其中的共有峰,利用中药色谱指纹图谱相似度评价系统(2012.130723版本)进行相似度评价,采用超高效液相色谱-四极杆飞行时间串联质谱法(UHPLC-Q-TOF-MS/MS)定性鉴别桔梗汤物质基准,并结合聚类分析(CA)、主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)等化学计量学方法筛选影响质量的主要差异性成分。通过测定1,1-二苯基-2-三硝基苯肼(DPPH)自由基的清除率来评价桔梗汤物质基准的抗氧化活性,并采用偏最小二乘回归分析方法建立桔梗汤物质基准中共有峰与抗氧化活性的谱效关系模型。结果显示,12批桔梗汤物质基准HPLC指纹图谱中得到7个共有峰,指认出3个成分(甘草苷、异甘草苷、甘草酸)均归属于甘草药材,相似度均大于0.900。采用UHPLC-Q-TOF-MS/MS鉴定出75个化学成分,其中13个化学成分归属于桔梗药材,57个化学成分归属于甘草药材。CA、OPLS-DA均将12批桔梗汤物质基准分为3类;PCA得到2个主成分,累计方差贡献率为92.001%;OPLS-DA得到4个影响桔梗汤物质基准整体质量的差异性成分,包含甘草酸和3种未知物。根据谱效关系模型可知,甘草酸和3种未知物是清除DPPH自由基的关键成分,直接影响桔梗汤物质基准的抗氧化活性。

       

      Abstract: To investigate the quality control method for Jiegeng decoction material benchmarks, the title research was conducted. With 12 batches of Jiegeng decoction material benchmarks as research object, high performance liquid chromatography (HPLC) fingerprints were established, and common peaks were identified by comparing with reference fingerprint. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (2012.130723 edition) was used for similarity evaluation. Ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry (UHPLC-Q-TOF-MS/MS) was employed for the qualitative identification of Jiegeng decoction material benchmarks. Chemometric methods including cluster analysis (CA), principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to find the main difference components affecting the quality. The antioxidant activity of the Jiegeng decoction material benchmarks was evaluated by determining the scavenging rate of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals, and the spectrum-effect relationship model between the common peaks in the Jiegeng decoction material benchmarks and antioxidant activity was established using partial least squares regression analysis method. As shown by the results, 7 common peaks were obtained from the HPLC fingerprints of 12 batches of Jiegeng decoction material benchmarks. 3 components (liquiritin, isoliquiritin, glycyrrhizic acid) were identified, all of which were attributed to Glycyrrhiza uralensis Fisch. herbs, and the similarity was greater than 0.900. 75 chemical components were identified by UHPLC-Q-TOF-MS/MS, of which 13 chemical components were attributed to Platycodon grandiflorus (Jacq.) A.DC. herbs and 57 chemical components were attributed to Glycyrrhiza uralensis Fisch. herbs. 12 batches of Jiegeng decoction material benchmarks were classified into 3 categories by CA and OPLS-DA. 2 principal components were obtained by PCA, with cumulative variance contribution rate of 92.001%. 4 differential components that effect the overall quality of the Jiegeng decoction material benchmarks were obtained by OPLS-DA, including glycyrrhizic acid and 3 unknown substances. The spectrum-effect relationship model indicated that glycyrrhizic acid and 3 unknown substances were the key components for scavenging DPPH free radicals, and the antioxidant activity of the Jiegeng decoction material benchmarks was directly affected by the above 4 chemical compounds.

       

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