Dingkun Dan (DKD), a reputable traditional Chinese medicine formula, has been used to treat gynecological diseases and showed significant clinical effects since ancient times. However, the application and development of DKD are seriously hampered by the unclear active substances. Structural characterization of compounds absorbed in vivo and their corresponding metabolites is significant for clarifying the pharmacodynamic material basis. In this study, an integrated strategy using ultra-performance liquid chromatography, coupled with quadrupole time-of-flight mass spectrometry and UNIFI™ software, was used to identify prototypes and metabolites after oral administration of DKD in rats. As a result, a total of 261 compounds, including 140 prototypes and 121 metabolites, were tentatively characterized in rat plasma, urine, and feces. The metabolic pathways of prototypes have been studied to clarify their possible transformation process in vivo. Moreover, an in vitro metabolism study was applied for verifying the metabolites under simulating the metabolic environment in vivo. This first systematic metabolic study of DKD is important for elucidating the metabolites and metabolic pathways and could provide a scientific basis for explaining the integrative mechanism in further pharmacology study.

中文翻译：

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超高效液相色谱-四极杆飞行时间质谱法研究中药定坤丹在大鼠体内外代谢的研究

定坤丹（DKD）是著名的中药方剂，自古以来就被用于治疗妇科疾病，临床疗效显着。然而，由于活性物质不明确，严重阻碍了DKD的应用和发展。体内吸收的化合物及其相应代谢物的结构表征对于阐明药效学物质基础具有重要意义。在这项研究中，使用超高效液相色谱法、四极杆飞行时间质谱法和 UNIFI™ 软件的综合策略用于鉴定大鼠口服 DKD 后的原型和代谢物。结果，在大鼠血浆、尿液和粪便中初步鉴定了总共 261 种化合物，包括 140 种原型和 121 种代谢物。已经研究了原型的代谢途径以阐明它们在体内可能的转化过程。此外，在模拟体内代谢环境的情况下，应用体外代谢研究验证代谢物。这项首次系统性的DKD代谢研究对于阐明代谢物和代谢途径具有重要意义，可为进一步药理学研究中解释整合机制提供科学依据。