基于超分子体系探究矿物药龙骨在含柴胡的角药配伍中的作用机制

Compatibility Mechanism of Mineral Medicine Os Draconis in Bupleuri Radix-containing Tri-herbal Medicines Based on Supramolecular Systems

目的:以龙骨、柴胡、牡蛎这组角药配伍入手,从超分子体系形成的角度初步探索矿物药龙骨在含植物药柴胡的角药配伍中的作用。方法:①观察龙骨、柴胡、牡蛎3组单煎液和柴胡-龙骨、柴胡-龙骨-牡蛎、柴胡-牡蛎3组共煎液的外观、丁达尔现象,测定各水煎液中混悬颗粒的平均粒径、分散性系数及Zeta电位,以扫描电镜(SEM)观察超分子结构微观形貌。②测定不同配伍系统pH、液体黏滞系数、液体表面张力、冻干粉得率等物理性质,结合红外吸收光谱法(FTIR)、紫外-可见光分光光度法(UV-Vis)检测不同配伍系统内的相互作用。③运用高效液相色谱法(HPLC)和超高压液相色谱串联四级杆飞行时间质谱联用技术(UPLC-Q-TOF-MS)测定不同配伍系统成分及含量的差异。结果:与单煎液相比,共煎液具更明显浑浊和丁达尔现象,共煎混悬液中的粒子变小,分布更均匀,且Zeta电位降低,体系更稳定,SEM下可见柴胡呈不规则层片状,柴胡-龙骨和柴胡-龙骨-牡蛎以球形纳米颗粒为主,柴胡-牡蛎则呈现不规则层片状和少量球形纳米颗粒混合的微观形貌特征;以单煎柴胡为基准,配伍共煎液的pH整体上升,黏度系数有所增加、液体表面张力有所降低,共煎液柴胡-龙骨冻干粉得率最高,其次为柴胡-牡蛎、柴胡-龙骨-牡蛎,而柴胡单煎液得率最低;FTIR主要变化为-OH的伸缩振动,共煎液明显向低频方向移动;UV-Vis显示,各组均在295.8 nm出现最大吸收,而吸光强度不同(柴胡-龙骨>柴胡-龙骨-牡蛎>柴胡-牡蛎>柴胡);以植物药柴胡的成分为指标,HPLC测定的甲醇提取成分的含量高于水煎提取,柴胡单煎液中主要为柴胡皂苷a(SSa)和柴胡皂苷c(SSc),共煎配伍后含量略高;UPLC-Q-TOF-MS在单煎液和共煎液中均可鉴定出37个化合物成分,种类没有变化。结论:龙骨、柴胡、牡蛎这组角药经共煎配伍后可形成粒径更小、更为均匀、稳定的纳米超分子体系,有助于柴胡主要成分的溶出,且含龙骨的配伍组在此过程中贡献最大。

Objective:By starting with the combination of Os Draconis,Bupleuri Radix,and Ostreae Concha,the role of mineral medicine Os Draconis in the combination of the Bupleuri Radix-containing tri-herbal medicines was preliminarily explored from the perspective of supramolecular system formation.Method:①The appearance and Tyndall phenomenon of single decoction of Os Draconis,Bupleuri Radix,and Ostreae Concha,as well as co-decoction of Bupleuri Radix-Os Draconis,Bupleuri Radix-Os Draconis-Ostreae Concha,and Bupleuri Radix-Ostreae Concha were observed,and the average particle size,dispersion coefficient,and Zeta potential of suspension particles in each decoction were determined.The micromorphology of supramolecular structures was observed by scanning electron microscope(SEM).②The pH of different compatibility systems,liquid viscosity coefficient,liquid surface tension,freeze-dried powder yield rate,and other physical properties were determined,and the interaction of different compatibility systems was detected by infrared absorption spectroscopy(FTIR)and UV-visible spectrophotometry(UV-Vis).③The composition and content difference of different compatible systems were determined by high-performance liquid chromatography(HPLC)and ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-QTOF-MS).Result:Compared with the single decoction,the co-decoction had more obvious turbidity and Tyndall phenomenon.The particles in the co-decoction suspension were smaller and more evenly distributed,and the Zeta potential was reduced,indicating a more stable system.Under SEM,Bupleuri Radix was irregularly lamellar,and Bupleuri Radix-Os Draconis and Bupleuri Radix-Os Draconis-Ostreae Concha were mainly spherical nanoparticles.Bupleuri Radix-Ostreae Concha was irregularly lamellar,with a small number of spherical nanoparticles.The pH of the single decoction of Bupleuri Radix and co-decoction increased,and the viscosity coefficient increased.The liquid surface tension decreased.The freeze-dried powder yield rate of the Bupleuri Radix-Os Draconis co-decoction was the highest,followed by Bupleuri Radix-Ostreae Concha decoction and Bupleuri Radix-Os Draconis-Ostreae Concha decoction,and the yield rate of Bupleuri Radix single decoction was the lowest.The main change of FTIR was the stretching vibration of-OH,and the codecoction moved to the low-frequency direction obviously.UV-Vis showed that the maximum absorption occurred at 295.8 nm for all groups,and the absorption intensity was different(Bupleuri Radix-Os Draconis>Bupleuri Radix-Os Draconis-Ostreae Concha>Bupleuri Radix-Ostreae Concha>Bupleuri Radix).The components of Bupleuri Radix were used as the indexes,and the content of methanol extract determined by HPLC was higher than that of water extract,and the components of Bupleuri Radix single decoction were mainly saikosaponin a(SSa)and saikosaponin c(SSc),which were slightly higher after co-decoction compatibility.UPLC-Q-TOF-MS could identify 37 compounds in both single decoction and co-decoction.Conclusion:The combination of Bupleuri Radix,Os Draconis,and Ostreae Concha can form a smaller,more uniform,and stable nano-sized supramolecular system,which is conducive to the dissolution of the main components of Bupleuri Radix,and the Os Draconis contributes the most in this process.