基于UHPLC-LTQ-Orbitrap-MS/MS技术的小儿豉翘清热颗粒化学成分鉴定及网络药理学研究

Identification of chemical constituents of Xiaoer Chiqiao Qingre Granules based on UHPLC-LTQ-Orbitrap-MS/MS and network pharmacology analysis

应用UHPLC-LTQ-Orbitrap-MS/MS技术对小儿豉翘清热颗粒中的化学成分进行定性分析,明确其物质基础,同时结合其入血成分,整合网络药理学研究方法探讨小儿豉翘清热颗粒潜在的作用机制。选用ACQUITY UPLC HSS T3(2.1 mm×100 mm,1.8μm)色谱柱和0.1%甲酸水溶液(A)-乙腈(B)流动相系统进行梯度洗脱;高分辨液质联用质谱正负离子扫描模式;根据精确相对分子质量数据和多级质谱碎片离子,结合对照品比对以及文献报道,共从小儿豉翘清热颗粒中分析鉴定了124个化学成分,其中有29个化合物经对照品比对完全确定。并借助UHPLC-LTQ-Orbitrap-MS/MS技术阐明小儿豉翘清热颗粒的主要入血成分,利用BATMAN-TCM和SwissTargetPrediction数据库对入血成分进行靶点预测,结合Cytoscape 3.8.2绘制关联网络图,对核心靶基因进行GO以及KEGG通路富集分析,构建成分-靶点-通路网络图,得到8个小儿豉翘清热颗粒治疗小儿发热的主要靶点,以及关键的8个入血成分,甘草次酸、橙皮素、大黄素、reticuline、大豆苷元、magnolignanC、magnolignan A、magnaldehyde D。推断小儿豉翘清热颗粒可能通过肿瘤坏死因子、PI3K-AKT等信号通路,改善消化道系统异常、炎症反应,氧化应激和内分泌紊乱等。该研究全面地阐明小儿豉翘清热颗粒中的化学成分,明确了主要入血成分同时整合网络药理学探讨其潜在机制,为小儿豉翘清热颗粒的临床应用提供了一定的理论参考。

This study aimed to qualitatively analyze the chemical components in Xiaoer Chiqiao Qingre Granules(XRCQ)by UHPLC-LTQ-Orbitrap-MS/MS and identify its material basis.The absorbed components in plasma were combined for exploring the potential action mechanism by integrated network pharmacology.ACQUITY UPLC HSS T3(2.1 mm×100 mm,1.8μm)column and mobile phase system of 0.1%formic acid solution(A)-acetonitrile(B)were used for gradient elution,followed by high resolution liquid chromatography-mass spectrometry in both positive and negative ion scanning modes.According to the precise relative molecular mass and MS/MS fragment ions,a total of 124 chemical components were identified in XRCQ by the comparison with references and literature reports,among which 29 compounds were completely confirmed by comparison with reference substances.Then,the main absorbed components of XRCQ in plasma were also analyzed and clarified by UHPLC-LTQ-Orbitrap-MS/MS.BATMAN-TCM and SwissTargetPrediction were used for target prediction of absorbed components in plasma.Following the plotting of association network with Cytoscape 3.8.2,the core targets were subjected to GO and KEGG pathway enrichment analysis and a component-target-pathway network was constructed.A total of eight main targets of XRCQ against fever in children were obtained together with eight absorbed components in plasma,including glycyrhydinic acid,hesperidin,emodin,reticuline,daidzein,magnolignan C,magnolignan A,and magnolaldehyde D.It was inferred that XRCQ might improve alimentary system abnormality,inflammatory response,oxidative stress,and endocrine disorder through tumor necrosis factor,PI3 K-AKT,and other signaling pathways.The present study comprehensively expounded the chemical profiles of XRCQ and the main absorbed components in plasma and predicted the potential mechanism of XRCQ based on integrated network pharmacology,which has provided certain theoretical reference for the clinical application of XRCQ.