Xu Lu-Lu, Liu Bin, Wang Fei, Gao Xiao-Hui, Wang Yu-Qi, Wang Hong-Jing, Li Ning, Zhang Jia-Yu
School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 102488, China.
KPC Pharmaceuticals, Inc., Kunming 650100, China.
Zhongguo Zhong Yao Za Zhi. 2018 Nov;43(22):4534-4540. doi: 10.19540/j.cnki.cjcmm.2018.0120.
Ultra-high-performance liquid chromatography coupled with tandem high-resolution mass spectrometry (UHPLC-HR-MSn ) 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.
采用超高效液相色谱-串联高分辨质谱(UHPLC-HR-MSn)法分析复方甘草片及其大鼠血浆中的主要代谢产物。在口服复方甘草片前后采集大鼠血浆。经固相萃取后,使用ACQUITY UHPLC BEH C₁₈(2.1 mm×100 mm,1.7 μm)色谱柱,以0.1%甲酸-乙腈溶液为流动相进行梯度洗脱。采用配备电喷雾离子源(ESI)的LTQ-Orbitrap质谱仪在正离子模式下分析复方甘草片中的化学成分及其在血浆样品中的原型和代谢产物。基于精确质量测定、保留时间和质谱裂解模式,从复方甘草片中初步鉴定出55种化合物,包括42种黄酮类、9种三萜类和4种生物碱。此外,还分析了大鼠口服复方甘草片后血浆中的代谢产物。共鉴定出26种化合物,包括20种原型和6种代谢产物,主要通过羟基化、葡萄糖醛酸结合和硫酸结合等代谢途径产生。结果表明,超高效液相色谱-线性离子阱四极杆Orbitrap高分辨质谱联用(UHPLC-LTQ-Orbitrap)技术能够全面阐明复方甘草片的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的代谢过程和药效物质基础提供科学依据。