Discovery of Cardioprotective Markers in Huangqi Shengmai Yin via Integrated UHPLC-Q-TOF-MS/MS Fingerprinting, Chemometric Quality Assessment, and Bioactivity Correlation.

Huangqi Shengmai Yin (HQSMY) displays notable cardioprotective properties. Nevertheless, the principal pharmacodynamic constituents underpinning this efficacy have not yet been fully clarified. To tackle this issue, an integrative workflow coupling UHPLC‑Q‑TOF‑MS/MS fingerprinting, chemometrics, and spectrum‑effect correlations was constructed for the systematic discovery of cardioprotective markers. Fifty-one components from 10 batches of HQSMY samples were characterized by UHPLC‑Q‑TOF‑MS/MS in conjunction with the GNPS molecular networking platform. Subsequently, chromatographic fingerprints were established, with 22 and 29 characteristic common peaks assigned in negative and positive ion modes, respectively. Interbatch consistency was assessed via chemometric methods. Spectrum‑effect relationship analyses were conducted through Pearson correlation, gray relational analysis, and OPLS regression, revealing that formononetin, astrapterocarpan, astragaloside II, schisandrin, and biochanin A exhibited highly significant positive correlations with H9c2 cardiomyocyte protective activity. All these compounds attained gray relational scores exceeding 0.7, implying strong associations with cardioprotective action. These five components were then chosen from the candidate markers for subsequent functional validation using the CCK-8 assay, which yielded EC50 values of 18.60 ± 3.15 µM (biochanin A), 26.42 ± 4.78 µM (formononetin), 92.93 ± 19.14 µM (astragaloside II), 70.85 ± 13.26 µM (schisandrin), and 72.73 ± 14.05 µM (astrapterocarpan). The cardioprotective effect of astragaloside II, schisandrin, and astrapterocarpan in H2O2-induced H9c2 injury model were reported for the first time. This integrated analytical strategy not only pinpoints the primary cardioprotective components of HQSMY but also furnishes a dependable methodological framework for quality evaluation and pharmacodynamic material research.