Abstract：
A reliable method using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was established to conduct a comprehensive analysis of the chemical constituents of Du-zhi pill (DZP) as well as their metabolites in rat plasma, urine and feces after gastric perfusion. The efficient on-line mass data acquisition modes combined the various off-line mass data mining strategy was applied. A full mass scan was performed, and then accurate MS/MS datasets were obtained through the use of a multiple mass defect filter (MMDF) and dynamic background subtraction (DBS)-dependent data acquisition method. Furthermore, post-acquisition data processing was conducted using various data-mining tools, including extracted ion chromatography (XIC), mass defect filtering (MDF), product ion filtering (PIF), and neutral loss filtering (NLF) (MetabolitePilot™). Finaly, a total of 176 compounds were identified or tentatively characterized in DZP. Moreover, a total of 233 components in vivo, which includes 92 prototype components and 141 metabolites, were unambiguously or tentatively identified in rat plasma, urine and feces. The metabolic pathways, including phase I reactions (hydroxylation, dehydroxylation and hydrogenation) and phase II reactions (acetylation, sulfation, glucuronidation and methylation), for the absorbed constituents, were explored and summarized. This is the first systematic study on the components of DZP and their metabolites in vivo. This study provide a valid analytical strategy for the characterization of chemical compounds and metabolites of TCM formulas. Moreover, an integrative strategy was proposed for the characterization and identification of chemical constituents and metabolites for additional TCM prescriptions.
摘要：
建立了一种可靠的超高效液相色谱-四极杆飞行时间质谱（UPLC-Q-TOF-MS）方法，对杜氏丸（DZP）的化学成分进行了综合分析。大鼠血浆中的代谢产物，胃灌注后的尿液和粪便。有效的在线海量数据采集模式结合了各种离线海量数据挖掘策略。进行了全质量扫描，然后通过使用多重质量缺陷过滤器（MMDF）和动态背景减除（DBS）相关的数据采集方法获得准确的MS/MS数据集。此外，采集后的数据处理是使用各种数据挖掘工具进行的，包括萃取离子色谱法(XIC)，质量缺陷过滤(MDF)，产品离子过滤（PIF），和中性损耗滤波(NLF)(MetabolitePilot™)。最后,在DZP中鉴定或初步表征了总共176种化合物。此外，体内共有233种成分，其中包括92种原型成分和141种代谢物，在大鼠血浆中明确或初步鉴定，尿液和粪便。代谢途径，包括第一阶段反应(羟基化，脱羟基和氢化)和第二阶段反应(乙酰化，硫酸化，葡糖醛酸化和甲基化)，对于吸收的成分，进行了探索和总结。这是首次对DZP及其代谢产物的体内成分进行系统研究。本研究为中药方剂化合物和代谢物的表征提供了有效的分析策略。此外，提出了一种综合策略来表征和鉴定其他中药处方的化学成分和代谢物。
