Objective Based on the supramolecular of traditional Chinese medicine (STCM) theory, this study systematically explored the material basis of Huangqi (Astragali Radix)-Ezhu (Curcumae Rhizoma) (AC) herb pair for synergistic anti-liver cancer, aiming to reveal the scientific connotation of its compatibility. Methods The STCM were separated by the centrifugation-dialysis method. The particle size and morphology were characterized by dynamic light scattering (DLS) technology and transmission electron microscopy (TEM), and the formation mechanism was analyzed by spectroscopic technology. The chemical composition was further analyzed by liquid chromatography-mass spectrometry (LC-MS). Its anti-liver cancer activity in vitro was evaluated. Results The results showed that STCM existed in both the AC co-decoction and the physically mixed solution, but the STCM in the co-decoction (AC-STCM) had smaller particle size, more uniform distribution, and higher stability; its self-assembly process may be jointly driven by components such as calycosin, formononetin, and demethoxycurcumin through hydrogen bonding and π-π stacking interactions. AC-STCM exhibited significant inhibitory effects on HepG2 cells (P < 0.01), with its in vitro antitumor activity notably superior to that of the co-decoction and the physically mixed STCM (Mix-STCM) (P < 0.01), while the co-decoction itself also showed better antitumor effects than the physical mixture (P < 0.01). Conclusion During the decoction process, AC herb pairs self-assemble through non-covalent bonds to form a stable STCM system, which constitutes an important material basis for their synergistic anti-liver cancer effects. This study is the first to explain the scientific connotation of AC for synergistic anti-liver cancer from the perspective of STCM, which provides a theoretical basis and experimental basis for the modernization of the theory of compound compatibility of traditional Chinese medicine.

supramolecules of traditional Chinese medicine  /  Astragali Radix  /  Curcumae Rhizoma  /  herb pairs  /  compatibility  /  material basis  /  LC-MS  /  calycosin  /  formononetin  /  demethoxycurcumin  /  antitumor activity