目的 以黄连厚朴汤（Huanglian Houpu Decoction，HHD）煎液种类、含量、相态为基础，研究配伍合煎的化学成分及含量变化；探究配伍合煎的组分自组装行为。方法 采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法（UPLC-Q-Exactive Orbitrap HRMS）鉴定并归属HHD化学成分；利用超速离心和透析技术对HHD进行相态拆分，使用马尔文粒径仪、透射电子显微镜对各相态的粒径、ζ电位、微观形态进行表征，选用紫外分光光度法、HPLC法对各相态的蛋白、多糖及黄连、厚朴有效成分表小檗碱、黄连碱、巴马汀、小檗碱、和厚朴酚、厚朴酚含量进行测定。结果 鉴定并归属HHD中的43种成分；分析其中2个未知化合物的裂解规律，发现可能存在组分自组装行为；对比合煎液、混合液及单煎液化学成分及含量，发现6种有效成分含量均表现为合煎液低于混合液低于单煎液；通过相态拆分获得5个亲水型相态，粒径分布在20～1 000 nm，主要为沉淀相态、真溶液相态和胶体相态；对比合煎液与混合液不同相态，分析蛋白、多糖、黄连与厚朴中6种有效成分含量，发现它们在沉淀相态中的含量均表现为合煎液低于混合液，而在胶体相态中正好相反；说明蛋白、多糖及6种有效成分通过配伍合煎后，可能发生结合，未沉降而保留在胶体相态，进一步推测组分自组装行为的存在。结论 HHD配伍合煎的化学成分及含量存在明显差异，这可能与中药汤剂配伍合煎的复杂变化有关，为后续深入研究中药汤剂组分自组装提供依据。

Objective To study the chemical composition and content changes of the decoction of Huanglian Houpu Decoction (HHD, 黄连厚朴汤) based on the decoction type, content and phase state, and to investigate the self-assembly behavior of the components of the decoction of HHD. Methods The compositions of HHD were identified by UPLC-Q Exactive Orbitrap HRMS. The phases of HHD were separated using ultracentrifugation and dialysis, and the particle size, ζ potential, and microscopic morphology of each phase were characterized by Zetasizer Nano ZS instrument and transmission electron microscope. The contents of proteins, polysaccharides, epiberberine, coptisine, palmatine, berberine, magnolol and honokiol in different phases of HHD were determined by UV spectrophotometry and HPLC. Results A total of 43 compositions were identified and assigned, the fragmentation patterns of two unknown compounds which revealed the possible existence of composition self-assembly behaviors were analyzed. Compared chemical compositions and contents of the combined decoction, the mixed decoction and the single decoction, it was found that the contents of six active compositions all showed combined decoction were less than mixed decoction, mixed decoction were less than single decoction. Five hydrophilic phases were isolated from HHD, the particle size distribution ranged from 20 to 1 000 nm, mainly including precipitation phases, true solution phase and colloidal phase. Compared different phases of combined decoction and mixed decoction, the contents of proteins, polysaccharides, and six active compositions from Coptis chinensis and Magnolia officinalis were analyzed, it was found that contents of these compositions in combined decoction were less than mixed decoction under the precipitation phases, while it was just the opposite under the colloidal phase. It indicated that during the compatibility of combined decoction, proteins, polysaccharides and six active compositions may combine with each other, remain in the colloidal phase without sedimentation, and further speculating the existence of self-assembly behaviors among the compositions. Conclusion There were significant differences in chemical compositions and contents during the compatibility of combined decoction in HHD, which may be related to the complex changes during the compatibility of combined decoction and providing a basis for the subsequent research on the composition self-assembly in traditional Chinese medicine decoctions.

R283.6

成都中医药大学西南特色中药资源国家重点实验室开放研究基金资助项目（SKLSCMR2021008）