A multidimensional strategy for exploring quality markers of Xue-Fu-Zhu-Yu decoction and its three processed products based on intestinal absorption and pharmacodynamics.

Ethnopharmacological relevance: Xue-Fu-Zhu-Yu (XFZY) formulas are well-known traditional Chinese medicine extensively utilized to treat various blood stasis syndromes (BSS) by promoting blood circulation and eliminating blood stasis. However, their quality control remains incomplete, probably leading to fluctuations in product quality and thus causing unsatisfactory therapeutic effects.

Aim of the study: To establish a multidimensional strategy that integrates chemical analysis, intestinal absorption, and pharmacodynamic effects for exploring Q-markers of XFZY formulations.

Materials and methods: The fingerprint spectrum analysis of four XFZY formulation intermediates and their component identification were conducted to provide chemical dimensions. The intestinal absorption experiment with everted gut sac model was used to obtain absorbable prototype components, establishing a link between chemical and absorptive dimensions. Blood-activating effects of four XFZY formulation intermediates in BSS models were evaluated, with candidate Q-markers acquired via Pearson's correlation analysis with the pharmacodynamic and absorption outcomes, completing the chemical-absorptive-pharmacological framework. The molecular docking, plasma recalcification time assays, and platelet aggregation experiments were performed to further confirm the final Q-markers for XFZY formulations.

Results: The fingerprints of XFZY formulation intermediates were established, resulting in 30 common peaks with identified chemical structures. 23 of these common compounds were found to be absorbed through the predominant absorption site of the jejunum. Subsequently, the ameliorative effects of XFZY formulas on body weight, hemorheology, NO level, and lipid levels in BSS model rats were confirmed, and then 14 potential Q-markers were obtained via Pearson's correlation analysis with pharmacodynamic data and absorbable components. Finally, gallic acid, neochlorogenic acid, hydroxysafflor yellow A, albiflorin, ferulic acid, 20-hydroxyecdysone, neohesperidin, meranzin, glycyrrhizic acid, and nobiletin were verified as the Q-markers of XFZY formulations by molecular docking and anticoagulant assays.

Conclusions: This study successfully constructed a chemical-absorptive-pharmacological three-dimensional strategy, identifying 10 Q-markers for XFZY formulations, which provides an important basis for constructing a comprehensive quality control system for XFZY formulations.