Multicomponent tracking and targeting strategy based on ion identity molecular networking for in vivo characterization of Weifuchun constituents

Background: Traditional Chinese medicine (TCM) demonstrates significant therapeutic efficacy in clinical practice; however, its complex composition and multicomponent synergistic effects present considerable challenges for research. Currently, efficient methods for identifying in vivo -exposed components of TCM and elucidating its pharmacologically active substances remain limited, which restricts a comprehensive understanding of its holistic mechanisms of action. Objective: To develop an integrated multicomponent tracking and targeting strategy based on ion identity molecular networking for targeted in vivo tracking and analysis of TCM components, using the proprietary multi-herb Chinese formula “Weifuchun” as a case study. Results: A total of 296 chemical constituents in Weifuchun were successfully identified. By integrating ultra-high-performance liquid chromatography-electrospray ionization-Orbitrap-mass spectrometry, ion identity molecular networking, and the Global Natural Products Social platform, components absorbed into the blood and distributed across multiple tissues in rats with chronic atrophic gastritis were detected, and their organ-specific in vivo distribution patterns were preliminarily characterized. This strategy enabled systematic tracing of in vivo -exposed components and their tissue distribution in the disease model, providing a basis for elucidating the multi-organ therapeutic characteristics of TCM. Conclusion: This study proposes a standardized and efficient integrated multicomponent tracking and targeting strategy for in vivo tracking of TCM constituents. It provides a novel methodological framework for investigating the in vivo processes and multi-organ synergistic mechanisms of herbal formulas, thereby supporting the modernization and evidence-based application of TCM.