目的 探究甘草酸与芫花素配伍后的自组装行为及分子缔合规律，从微观层面分析甘草Glycyrrhizae Radix et Rhizoma-芫花Genkwa Flos配伍禁忌与成分状态的相关性。方法 通过动态光散射（dynamic light scattering，DLS）、扫描电子显微镜（scanning electron microscope，SEM）和超滤分离，分析甘草酸-芫花素配伍溶液中微粒的粒径、形貌和超滤分离行为。结果 甘草酸与芫花素以55∶1和45∶1的质量浓度比配伍后，粒径由相应质量浓度的甘草酸单体溶液的100～150 nm显著减小至约50 nm，且配伍后DLS在10～100 nm出现了单体溶液没有的光强信号。SEM观察到甘草酸-芫花素配伍溶液形成了规整的立方体颗粒，与单体的球状、长针状完全不同。使用截留相对分子质量分别为3 000、10 000、30 000、50 000、100 000的超滤膜进行分离时发现，随着甘草酸质量浓度升高，甘草酸透过率逐渐下降，透过率10%时的截留相对分子质量（T₁₀）与缔合系数（N）逐渐增大。在同质量浓度条件下，配伍体系中甘草酸透过率大于甘草酸单体体系，T₁₀和N小于单体体系。此外芫花素在含甘草酸体系中透过率显著提高。药材饮片煎煮体系与单体体系规律一致。对比单煎液与合煎液的SEM，甘草单煎液的颗粒结构规则，尺寸相对均一，芫花单煎液颗粒尺寸差异大。合煎液未能重现单体配伍的规整立方体，但颗粒均一性介于2种单煎液之间。结论 甘草酸与芫花素在水溶液中并非简单共存，芫花素以分子替换的形式进入甘草酸胶束，自组装形成新的缔合物，为研究甘草-芫花药对“反”的机制提供了新思路。

Objective To explore the self-assembly behavior and molecular association patterns of glycyrrhizic acid (GA) and genkwanin (Gen) after combination, and to analyze the correlation between the incompatibility and the molecular state of Gancao (Glycyrrhizae Radix et Rhizoma)-Yuanhua (Genkwa Flos) from the microscopic perspective. Methods Employ dynamic light scattering (DLS), scanning electron microscopy (SEM), and ultrafiltration to characterize the particle size, morphology and ultrafiltration behavior of GA-Gen mixed solutions. Results When GA and Gen were combined at mass ratios of 55:1 and 45:1, the particle size decreased significantly from 100—150 nm (GA monomer solutions) to approximately 50 nm. After combination, DLS showed a light intensity signal that was not present in the monomer solution at 10—100 nm. SEM images showed that the GA-Gen compatible solutions formed well-defined cubic particles, distinctly different from the globular and long needle-like morphologies of the individual components. When ultrafiltration membranes with molecular weight cut-off (M_w) of 3 000, 10 000, 30 000, 50 000 and 100 000 were used for separation, it was found that with the increase of GA concentration, the transmittance of GA gradually decreased, and the molecular weight at 10% transmittance (T₁₀) and the association coefficient (N) also gradually increased. At the same concentration, the transmittance of GA in the compatibility system was higher than that in the GA monomer system, and T₁₀ and N were lower than those in the monomer system. In addition, the transmittance of Gen in GA-containing system was significantly improved. The decoction system of medicinal materials is consistent with the trends of monomer system. Comparing the SEM of single decoction and co-decoction, the particle size of Glycyrrhizae Radix et Rhizoma single decoction was regular and the size was relatively uniform, and the particle size of Genkwa Flos single decoction was greatly different. The co-decoction failed to reproduce the regular cube of monomer compatibility, but the particle uniformity was between the two single decoctions. Conclusion GA and Gen do not simply coexist in aqueous solution. Instead, Gen is incorporated into GA micelles via molecular substitution, leading to the formation of new self-assembled associates. These findings provide a novel perspective for understanding the mechanism of “reverse” of Glycyrrhizae Radix et Rhizoma-Genkwa Flos herb pair.

R283.6

国家自然科学基金资助项目（82274106）；中药制药过程控制与智能制造技术全国重点实验室创新项目（NZYSKL240207）；南京中医药大学中药学一流学科科学研究培育项目（ZYXPY2024-006）