基于活性导向和近红外光谱技术的丹参活血化瘀成分转化规律和调配研究

he salvianolic acids, the important components of Salvia miltiorrhiza Bge. to activate blood and resolve stasis, easy to degradated to each other during preparation, leads to significant variation of concentrations of salvianolic acids, especially the unstable proportion between Salvia acid B danshensu, which increased the instability of the quality and efficacy of chinese patent medicines originated from Salvia miltiorrhiza Bge. Thus, it’s urgently needed to understand the optimal ratio of the active ingredients and the transformation law of the salvianolic acids. And it’s necessary to carry out the control of the transformation process of the salvianolic acids. In this project, the salvianolic acids were considered as measuring object. Firstly, the fingerprint and the material basis of the salvianolic acids extract were carried out. Secondly, a model of blood stasis was established by giving rats adrenaline and iced water method, to screen the best proportional relationship among salvianolic acids of the salvianolic acids extract and the dose-effect relationship between the salvianolic acids extract and efficacy. Thus, the study of parameters and constraints of transformation law among salvianolic acids under different conditions, such as pH, reaction temperature, reaction time and concentrations of components, etc. were spread out by near-infrared spectroscopy combined with traditional Chinese medicine fingerprint technology, chemometrics technology, in order to reveal the transformation law of the salvianolic acids, and obtain the salvianolic acids extract of clear proportional relationship and the optimal effect by the allocation optimization technology. Finally, a preparation process of the salvianolic acids extract with a definite material basis and clear proportional relationship among the salvianolic acids, which could provide a base for the large scale production and depth development.

丹参活血化瘀功效主要成分为丹酚酸类。在制备过程中，丹酚酸类成分多相互转化，造成各种酚酸成分含量变异明显，特别是丹酚酸B和丹参素的比例关系不稳定，造成丹参制剂物质基础组成不稳定，同时多种酚酸类成分之间存在拮抗或促进作用，成分组成的不稳定性对丹参制剂的实际疗效影响极大。因而亟需研究丹参活血化瘀的最优活性成分比例，并明确其相互转化规律，并实现转化过程实时控制，以期实现丹参制剂的质量与疗效双重稳定。本项目前期已明确丹参指纹图谱和物质基础，拟进一步采用皮下注射肾上腺素-冰水法制备大鼠急性血瘀模型，确定药效最优的丹酚酸类成分的含量比例关系。然后采用近红外光谱技术，开展不同条件下（pH、温度、时间、浓度等）丹酚酸类成分的转化参数、约束条件等研究，揭示丹酚酸类成分的转化规律，辅以最优化调配技术制备预期提取物。最终实现丹参活血化瘀成分制备工艺的实时控制，为丹参制剂深度开发、大规模生产等提供研究基础。

丹参活血化瘀功效主要成分为丹酚酸类。在制备过程中，丹酚酸类成分多相互转化，造成各种酚酸成分含量变异明显，特别是丹酚酸B和丹参素的比例关系不稳定，造成丹参制剂物质基础组成不稳定，同时多种酚酸类成分之间存在拮抗或促进作用，成分组成的不稳定性对丹参制剂的实际疗效影响极大。.鉴于此，本研究建立了丹参水溶性成分指纹图谱，鉴定了丹参水提液中20种化学成分；考察了急性血瘀与急性寒凝血瘀大鼠动物模型的建模方法，并开展了不同剂量丹参水溶性提取物的活血化瘀药效研究，通过对5个指标成分采用均匀设计并进行逐步回归分析，得到最优的指标成分配伍比例（丹参素：原儿茶醛：咖啡酸：迷迭香酸：丹酚酸B的最优量效比例为486：41：173：89：18144，其中丹参素、丹酚酸B为主要核心指标），并进行效果验证研究。同时初步开展了丹参治疗急性血瘀证的代谢组学研究和网络药理学研究，对丹参活血化瘀机制做了初步探索研究，发现血瘀证与氨基酸代谢、脂质代谢、核苷酸代谢、能量代谢、炎症反应等密切相关。丹参活血化瘀相关的Q-marker为丹参素、原儿茶醛、迷迭香酸和丹参酚酸B等成分。.开展了丹酚酸B的提取工艺、纯化工艺研究。在最优比例指标成分比例下，以关键核心成分丹酚酸B和丹参素为指标，以及丹酚酸B转化为丹参素工艺研究，并采用近红外光谱技术对提取过程和转化过程开展了在线检测研究，预测效果良好。.开发了最优化调配算法，收集22个批次浓度的丹参提取液，设定目标成分比例关系稳定（丹参素：丹酚酸B=1：37）为约束条件，并选择其中2个和5个批次为例进行最优比例的调配研究，计算其调配比例，制备出组成成分确定、含量比例关系稳定的丹酚酸提取物（提取液）。.本研究初步阐明了丹参活血化瘀的药效物质基础，提供了一套物质基础明确、疗效确切、工艺稳定的丹酚酸类提取物制备工艺。本研究为丹参物质基础、质量标准、药效研究、生产过程控制及相关中药大品种二次开发及质量标准提升等提供了较好的研究基础。