目的 对不同煎煮时间的补阳还五汤（Buyang Huanwu Decoction，BHD）的汤液进行相态拆分和表征，探究汤剂煎煮过程中的相态变化及成分分布规律，为阐明中药复方成分协同起效的物质基础提供参考。方法 分别煎煮BHD 30、60、90、120 min制得汤液，采用“高速离心-透析”法分离得到混悬相态和纳米相态溶液；通过马尔文粒径仪、紫外/红外光谱、透射电子显微镜表征2种相态；测定BHD原汤及纳米相态、混悬相态的冻干粉得率，并采用液质联用仪（LC-MS/MS）测定BHD原汤及2种相态中15种指标成分含量。结果 4个煎煮时间点的BHD原汤、纳米相态、混悬相态均形成类球形颗粒，其中纳米相态的粒径随着煎煮时间的延长，由314.7 nm（30 min）降至170.2 nm（120 min）；多糖主要富集于纳米相态，而蛋白质主要富集于混悬相态；光谱分析结果显示，不同煎煮时间的BHD原汤和各相态的化学结构大致相似，但官能团的含量存在差异；LC-MS/MS结果显示，15种指标成分中多数成分在混悬相态中的平均含量高于纳米相态，其中黄芩苷、异亮氨酸、阿魏酸、腺嘌呤、丹皮酚、黄芪甲苷、毛蕊异黄酮、芒柄花素、山柰酚-3-O-芸香糖苷9种成分在各相态中的含量具有显著差异（P＜0.05）。结论 煎煮的热力学过程是驱动BHD成分自组装形成不同相态的关键因素。BHD成分在煎煮过程存在相互作用使药效成分在各相态间的含量呈现较大差异，为研究中药汤剂煎煮的科学性提供物质基础。

To investigate phase separation and characterize the decoction of Buyang Huanwu Decoction (BHD, 补阳还五汤) prepared at different decoction times, exploring phase transitions and component distribution patterns during the decoction process. This study aims to provide insights into the material basis for synergistic effects among components in traditional Chinese medicine formulas. Methods Decoction extracts were prepared by boiling BHD for 30, 60, 90, and 120 min. The extracts were separated into suspension and nano-phase solutions using the “high-speed centrifugation-dialysis” method. Both phases were characterized using a Malvern particle size analyzer, UV/IR spectroscopy, and transmission electron microscopy. The freeze-dried powder yield was determined for the original decoction and the nano- and suspension-phase solutions. The content of 15 target components in the original decoction and the two phases was measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results Spherical-like particles formed in the original decoction, nano-phase, and suspension phase at all four decoction time points. The particle size of the nano-phase decreased from 314.7 nm (30 min) to 170.2 nm (120 min) with prolonged boiling. Polysaccharides were predominantly enriched in the nano-phase, while proteins were mainly concentrated in the suspension phase. Spectral analysis revealed that the chemical structures of the original decoction and each phase were broadly similar across different decoction times, though functional group contents varied. LC-MS/MS results indicated that the average content of most components in the suspension phase exceeded that in the nano-phase. Among these, baicalin, l-isoleucine, ferulic acid, adenine, paeonol, astragaloside IV, calycosin, formononetin, and kaempferol-3-O-rutinoside exhibited significant content differences across phases (P < 0.05). Conclusion The thermodynamic process of decoction is the key factor driving the self-assembly of BHD components into distinct phases. Interactions among components of the BHD during boiling cause significant variations in the content of active ingredients across different phases, providing a material basis for studying the science of boiling Chinese herbal decoctions.

R283.6

国家自然科学基金资助项目(82274215);国家自然科学基金青年基金资助项目(82404815); 湖南中医药大学2022年度校级“揭榜挂帅”专项(2022-12-27-1)