Metabolites have a close relationship with the efficacy and safety of herbal medicines. However, ubiquitous matrix interferences, complex co-elution, and minor or trace amounts in plasma restrict the comprehensive identification of metabolites. In this study, an efficient strategy comprising a mass defect filter and time-staggered targeted ion lists was established to characterize the metabolites of alkaloids of Uncaria rhynchophylla (UR) for the systematic comparison of metabolic differences in rat, mouse, dog, pig, monkey and human liver microsomes. The mass defect filter model effectively decreased interfering ions by 63–68%, and time-staggered precursor ion lists significantly increased the number of triggered MS/MS fragmentation by 65–120% in liver microsomes of six species. Ultimately, a total of 165 metabolites in the liver microsomes of six species were tentatively characterized, and the main metabolic pathways were demethylation, isomerization, hydrolysis, oxygenation and dehydrogenation. The results showed that the mouse liver microsomes exhibited metabolic behavior most similar to human metabolism of UR alkaloids. We hope that these results provide basic data for further investigation of UR metabolism in different species, and that the strategy can provide a reference for metabolite characterization of herbal medicines in complex biological matrix.

代谢产物与草药的功效和安全性密切相关。但是，普遍存在的基质干扰，复杂的共洗脱以及血浆中微量或微量限制了代谢物的全面鉴定。在这项研究中，建立了一种有效的策略，该策略包括质量缺陷过滤器和错开时间的靶向离子列表，以表征钩藤Uncaria rhynchophylla生物碱的代谢产物。（UR）用于系统比较大鼠，小鼠，狗，猪，猴子和人肝微粒体的代谢差异。质量缺陷过滤器模型有效地将干扰离子减少了63–68％，并且时间交错的前体离子列表显着增加了6种物种的肝脏微粒体中触发的MS / MS碎片数量，增加了65–120％。最终，初步鉴定了六个物种的肝微粒体中的165种代谢物，其主要代谢途径为脱甲基，异构化，水解，氧化和脱氢。结果表明，小鼠肝微粒体表现出的代谢行为与人类UR生物碱的代谢最相似。我们希望这些结果为进一步研究不同物种的UR代谢提供基础数据，