基于UHPLC-Q-TOF-MS技术研究黄甲软肝颗粒及其中间体物质基础

Analysis of Chemical Constituents in Huangjia Ruangan Granules and Intermediate Based on UHPLC-Q-TOF-MS

目的:系统分析黄甲软肝颗粒及其中间体的化学成分,阐明其复方新药的化学物质基础。方法:采用超高效液相色谱-飞行时间串联质谱(UHPLC-Q-TOF-MS)技术,离子源为ESI,色谱条件:Agilent Zorbax Eclipse Plus C_(18)色谱柱(2.1mm×150mm,1.8μm),流动相为0.1%甲酸水溶液(A)-0.1%甲酸乙腈(B),洗脱梯度:0~20min,3%~60%B;20min~25min,60%~95%B;25min~30min,95%B;30min~31min,95%~3%B;31min~35min,3%B,流速0.35mL·min^(-1),柱温35℃,进样量1μL。质谱分析采用正、负离子模式扫描,扫描范围m/z100~1700。通过与自建化学成分数据库比对(部分对照品验证)后对各离子峰进行鉴定。结果:从黄甲软肝颗粒中共鉴定出126个化学成分,黄甲软肝中间体中共鉴定出141个化学成分,包括有机酸类、酚酸类、黄酮类、三萜类、皂苷类等。结论:建立的UHPLC-TOF-MS分析方法,结合数据库匹配,可全面快速地分析黄甲软肝颗粒及中间体中的化学成分,为其组方药效物质基础及质量控制研究提供科学依据。

Objective:In order to systematically clarify the chemical composition of Huangjia Ruangan granules,the main chemical components in this preparation and intermediate were rapidly identified and assigned.Methods:Ultra-performance liquid chromatography-time-of-flight mass spectrometry(UHPLC-Q-TOF-MS)was used.Agilent Zorbax Eclipse Plus C_(18) column(2.1mm×150mm,1.8μm)was employed for UHPLC analysis with the mobile phase of 0.1%formic acid aqueous solution(A)-0.1%formic acid acetonitrile(B)for gradient elution(0~20min,3%~60%B;20min~25 min,60%~95%B;25min~30min,95%B;30min~31min,95%~3%B;31min~35min,3%B).The flow rate was 0.35mL·min^(-1),the column temperature was 35℃,and the volume of sample injection was 1μL.Electrospray ionization(ESI)was applied for scanning under positive and negative ion modes with the scanning range of m/z100~1700.MS mode was used to collect mass spectral data.The ion peaks were identified by comparing with the information of control substances,literature references and self-built database.Results:A total of 126chemical components were identified in Huangjia Ruangan granules,and 141chemical components were identified in Huangjia Ruangan intermediate.These ingredients include organic acids,phenolic acids,flavonoids,triterpenes,saponins and other components.Conclusion:The established UHPLC-Q-TOF-MS can comprehensively and rapidly analyze the chemical constituents in Huangjia Ruangan granules and intermediate,which can lay a foundation for further research on the pharmacodynamic material basis and quality control of Huangjia Ruangan granules.