基于免疫层析技术的大黄真伪优劣同步快速分析方法研究

It is important to establish a simultaneous and rapid analysis method for identifying the authenticity and evaluating the quality of Traditional Chinese medicine (TCM) for promoting the development of TCM. Rhei Radix et Rhizoma, a widely used TCM, is one of major TCM breeds in the international market for earn foreign exchange through exports. In this project, rhapontin which used as the identification index of rhubarb will be coupled with carrier protein by active ester action. And a special monoclonal antibody against rhapontin will be produced. While, a hapten model of anthraquinones which served as the quality control marker will be established and a class-special monoclonal antibody against anthraquinones will be produced. Finally, a gold immunochromatography assay (GICA) which can simultaneous detect rhapontin and anthraquinones contents will be established. Moreover, GICA has many advantages such as simple, fast, no dependence of instrument, and so on. The main significance of this work is to provide a probable way for detecting compounds of same kind by GICA. Furthermore, the study will provide a reference method for simultaneous identifying the authenticity and evaluating the quality of TCM with the rapid and simple characteristics.

建立中药真伪优劣同步快速鉴定方法对保障中药质量，促进中药发展具有重要意义。大黄为临床常用中药，也是我国中药出口创汇的主要大品种之一。本项目以中药大黄为例，以大黄真伪鉴定指标性成分土大黄苷和质量优劣评价指标性成分蒽醌类成分为研究对象，通过活泼酯法合成土大黄苷完全抗原，制备抗土大黄苷特异性单克隆抗体；通过构建蒽醌类成分半抗原模型，制备“广谱”特异性识别蒽醌类成分的单克隆抗体，解决免疫层析技术检测一类化学成分的技术难题；以胶体金为标记物，制备胶体金标记抗体，并建立同时检测土大黄苷和蒽醌类成分总量的免疫检测方法，从而实现大黄真伪和品质优劣的同步、快速鉴定。本项目将为解决免疫层析技术检测一类化学成分的问题提供新思路，并为中药的真伪优劣同步、快速鉴定提供参考方法。

中药鉴定的主要任务是鉴定药材的真伪优劣，该技术的实际应用多在于中药材的种植基地、交易市场等产业前端或中药零售药店等，然而现有的中药客观鉴定方法多需要依赖于仪器分析，无法实现中药鉴定的现场化应用，且仪器分析通量较低，无法很好的满足高通量鉴别的实际需要。鉴于以上矛盾，本研究将免疫分析及免疫层析技术的简便、快速、高通量及可视化的优点与中药鉴定技术相结合，并以常用中药大黄为例，尝试建立中药大黄的免疫分析技术，为中药高通量、现场化鉴定技术的开发提供参考。本项目主要实现了：①建立了土大黄苷定性检测方法为大黄类药材的真伪鉴定提供了快速、高通量的鉴定方法；②建立了大黄酸定性定量检测方法为大黄类药材的质量评价提供了补充方法；③基于胶体金免疫层析技术建立了蒽醌类成分大黄酸和芦荟大黄素的同步半定量快速检测方法填补了大黄质量现场化客观评价的技术空白。项目执行期间，共发表相关学术论文6篇，其中SCI论文2篇，授权实用新型专利2项。本项目的研究成果将为中药现场化鉴定技术的开发提供有益参考，推动中药鉴定技术的多元化发展。