复方甘草片大鼠血浆移行成分的分析鉴定

Rapid characterization of chemical constituents and rat metabolites of Fufang Gancao Tablets by UHPLC-LTQ-Orbitrap mass spectrometer

采用UHPLC-HR-MSn技术分析复方甘草片原型成分及其在大鼠体内的代谢物。口服复方甘草片后,收集给药前后的大鼠血浆,经固相萃取进行处理后,采用ACQUITY UPLC BEH C18(2. 1 mm×100 mm,1. 7μm)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;应用LTQ-Orbitrap质谱正离子模式分析复方甘草片中的化学成分以及血浆样品中的原型成分和代谢产物。根据精确相对分子质量数据和多级质谱碎片,结合对照品比对,从复方甘草片中分析鉴定了55个成分,包括42个黄酮、9个萜类、4个生物碱;从大鼠血浆中分析检测了26个原型产物及代谢产物,主要代谢途径为羟基化、硫酸酯化、葡萄糖醛酸化等。应用UHPLC-LTQ-Orbitrap高分辨液质联用技术较为全面地阐明了复方甘草片化学成分及其在大鼠血浆中的移行成分,为进一步研究复方甘草片体内代谢过程以及药效物质基础提供了科学依据。

Ultra-high-performance liquid chromatography coupled with tandem high-resolution mass spectrometry(UHPLC-HRMSn) method was used to analyze the constituents of Fufang Gancao Tablets and its main metabolites in rat plasma. Rat plasma was collected both before and after oral administration of Fufang Gancao Tablets. After solid phase extraction,ACQUITY UHPLC BEH C18(2. 1 mm × 100 mm,1. 7 μm) was used with 0. 1% formic acid-acetonitrile solution as the mobile phase for gradient elution. The chemical components in Fufang Gancao Tablets and their prototypes and metabolites in plasma samples were analyzed by LTQ-Orbitrap equipped with an ESI ion source in a positive ion mode. Based on the accurate mass measurements,the retention time and mass fragmentation patterns,a total of 55 compounds were tentatively identified from Fufang Gancao Tablets,including 42 flavonoids,9 triterpenes and 4 alkaloids. Furthermore,metabolites in rat plasma after oral administration of Fufang Gancao Tablets were also analyzed. A total of 26 compounds were identified,including 20 prototypes and 6 metabolites mainly through metabolic pathways of hydroxylation,glucuronide conjugation,and sulfate conjugation,etc. Our results showed that the ultra-high-performance liquid chromatography coupled with linear ion trap quadrupole Orbitrap high resolution mass spectrometry(UHPLC-LTQ-Orbitrap) could comprehensively elucidate the chemical constituents of Fufang Gancao Tablets and their migrating components in rat plasma,providing scientific basis for further studying the metabolism process and pharmacodynamic substance of Fufang Gancao Tablets.