Identification of Quality Markers for Tongmai Qiangshen Medicinal Liquor in the Treatment of Heart Failure Based on Multicomponent Qualitative–Quantitative Analysis and Integrated Bioinformatics

ABSTRACT Heart failure (HF) is an end stage of cardiovascular diseases. Tongmai Qiangshen Medicinal Liquor (TMQSJ) is an over‐the‐counter traditional Chinese medicinal liquor, but its efficacy‐related material basis and mechanisms in HF remain unclear. In this study, UPLC‐Q‐TOF‐MS/MS was used to characterize TMQSJ constituents and prototype compounds in rat plasma. HPLC fingerprinting, spectrum‐effect relationship analysis, single‐compound activity evaluation, and integrative bioinformatics were combined to screen efficacy‐related candidate quality markers and HF‐associated candidate genes. A total of 223 compounds were characterized, including 33 prototype plasma components. Chemometric analysis identified caffeic acid, Epimedin A, Epimedin B, Epimedin C, and icariin as characteristic constituents. Spectrum‐effect analysis based on 10 batches of TMQSJ and validation in DOX‐injured AC16 cells highlighted Epimedin A, Epimedin B, and Epimedin C. Among them, Epimedin A and Epimedin C showed prototype plasma exposure and were prioritized as efficacy‐related candidate quality markers. Integration of predicted targets of prototype plasma components with HF transcriptomic data and machine‐learning analysis identified BCL2L1, STAT3, HIF1A, MAPK8, and ESR1 as five candidate core genes. Immune cell estimation and molecular docking provided computational clues. Overall, this study establishes an integrated framework linking chemical quality, pharmacological association, in vivo exposure, and mechanistic prediction for TMQSJ quality evaluation.