Dynamic stability-activity profiling of traditional Chinese medicine: Decoding functional substances through forced degradation characteristics using Guizhi Fuling capsules as a demonstration.

Traditional Chinese medicine (TCM) is a complex system with multiple components, and its effective ingredients are difficult to identify. This study aims to develop a dynamic stability-activity research strategy, taking Guizhi Fuling Capsules (GFC) as an example, to explore the stability of chemical components and decode functional substances through forced degradation experiments. GFC, a widely prescribed formula for gynecological blood stasis syndrome. Given the established association between dysmenorrhea pathogenesis and elevated cyclooxygenase-2 (COX-2) activity in endometrial cells, this study employed COX-2 enzyme inhibition as an in vitro bioassay to characterize the biological activity of GFC. Following the International Conference on Harmonization (ICH) guidelines, hydrolysis (acid, alkali, and neutral), oxidation (H2O2), thermal, and photolysis degradation tests were conducted. Significant degradation of major components, including total phenols, gallic acid derivatives, free fatty acids (linolenic/linoleic/oleic acid), paeonol, and cinnamaldehyde occurred under acidic, alkaline, oxidative, and thermal conditions. Crucially, Pearson correlation analysis revealed these components' stability positively correlated with preserved COX-2 inhibitory potency. Validation experiments further demonstrated that the total phenolic fraction and individual compounds (e.g., gallic acid) exhibited stronger COX-2 inhibition than intact GFC in vitro, suggesting total phenols, gallic acid, ethyl gallate, and 1,2,3,4,6-O-pentagalloyl glucose contribute significantly to GFC's bioactivity this study establishes a novel "stability-activity" assessment paradigm for traditional formulas and underscores the necessity of future in vivo investigations to elucidate integrated mechanisms within physiological contexts.