中空纤维/石墨烯/有机溶剂固-液协同单元阵列微萃取及其在中药活性成分分析中的应用

Building the separate and analysis methods on active components from Chinese herbal medicine, which meet herbal character, is a hotspot. For the analysis of low concentration active components from Chinese herbal medicine, a new extraction pattern that is cell array of hollow fiber/graphene/organic solvent synergistic microextraction has been proposed based on hollow fiber liquid phase microextraction and nanomaterial. In this work, the organic solvents contain functionalized graphene were filled into the pores of hollow fiber, and this special cell array as multivariate extraction system was formed. Under the optimal experimental conditions, the active components from Chinese herbal medicine were extracted and determinated by solid-liquid synergistic microextraction combined with chromatography technique. By comparing the difference about chemical composition and physical structure of cell array between before and after exctraction ,to research the special forces and interactions among graphene, extractant, hollow fiber and active components in Chinese herbal medicine, and then the extraction mechanism about the cell array of hollow fiber/graphene/organic solvent synergistic microextraction was elaborated. In this way, to determine partition coefficient and enrichment factor of the analyte, study the correlation relation between the above two-parameter and each physicochemical character on polynary system, and make clear their optimum synergistic action, and then establish the mathematical models in microextracting herbal active components and develop a series of analytical methods about herbal active components by the cell array of solid-liquid synergistic microextraction. In this study, we expatiated on the extraction mechanism about multivariate extraction system in a new viewpoint， and it has both scientific and practical significance on building the separate and analysis methods on active components from Chinese herbal medicine, which meet herbal character.

构建符合中药特点的活性成分分离、分析方法是中药现代研究的热点。中空纤维/石墨烯/有机溶剂固-液协同单元阵列微萃取是课题组在前期中空纤维液相微萃取研究基础上，结合纳米技术提出的一种新型萃取模式。本课题旨在将功能化石墨烯有机分散液填充于中空纤维壁孔中形成"固-液协同微萃取单元阵列"的多元萃取体系，并结合色谱技术对中药活性成分的萃取和浓缩进行研究。根据萃取前后单元阵列化学组成和物理结构的变化，探索单元阵列与目标分析物之间的特殊作用力，阐明固-液协同微萃取机理；通过分析单元阵列中各相性质参数与目标分析物的分配系数和浓缩倍数之间的相关关系，探讨各参数间的最佳协同萃取作用；在此基础上，构建中药活性成分的固-液协同微萃取分离优化模型，建立以多元萃取体系为核心的多种中药活性成分的分析检测方法。本课题从全新的视角阐述多元萃取模式的作用机理，对构建符合中药特点的活性成分分离、分析方法具有重要意义。

揭示中药多成分、多靶点的作用特点，阐明中药药效物质基础，探索体内外中药活性成分的分析检测策略是中药研究中的热点话题。.本项目以中药活性成分为主要研究对象，旨在构建多元萃取体系为核心的一系列中药活性成分的分析检测方法，并将其与药物活性筛选技术相结合，实现体内外活性成分的同步筛选和分析检测。.项目组以菲醌类、苯丙酸类、生物碱类、多酚类、黄酮类、苦参碱类、五环三萜类、香豆素类等8大类中药活性成分为模型化合物，以固液协同微萃取技术为核心，建立了14种中药活性成分的体外定量分析方法，优化了相关萃取条件如固相材料的性质、粒径大小、表面积及用量，萃取溶剂种类及用量，样品相体积、pH及离子强度，萃取时间，搅拌速度等因素，比较了相关萃取模式的差异，阐明萃取机理。.同时，将其中部分方法作为中药活性筛选后的体内定量测定方法，建立完整的体内外中药活性成分的定性定量同步分析方法。.项目组以中空纤维细胞或靶标捕获-色谱/质谱法为筛选技术对中药提取液中活性成分进行活性筛选，实验详细考察了种植癌细胞或载有靶标的中空纤维筛选前后细胞的存活率和死亡率、阳性和阴性对照试验、方法重现性和可靠性。实验对有明显保留且稳定的活性成分通过与其对照品保留时间比对，明确其中药固有成分群，初步研究了活性成分与癌细胞上的结合部位及位点；比较和讨论了活性成分与靶点的结合能力和作用特点，研究和测定了血药浓度、组织浓度分布和药代动力学情况。在此基础上，同步建立体内活性成分检测方法寻找真正起效的活性成分群，对比体外固有成分与体内真正起效的活性成分的相关关系，明确中药药效物质基础。.总之，本项目将固液协同微萃取方法与中空纤维细胞捕获-高效液相色谱法相结合可实现体外活性成分筛选和体内活性成分检测的同步分析，阐明药效物质基础，为中药个性化质量标准的制定提供了新思路和新方法。