基于MIPs-DNA适配体识别技术的甾体激素生产过程废液在线分析方法的建立与评价

Steroid hormones have strong biological activities, and the waste emission produced during production process poses a serious threat to ecology. However, the analysis and monitoring of the steroid hormones are still in its infancy. This proposal is aimed to solve the problems of the pollutant analysis in the steroidal drug production process, such as poor specificity, low sensitivity, lagged behind analysis process and a poor correlation between the analytical targets and the physiological activity. Based on the prophase work, this project is planned to prepare molecularly imprinted polymers with high adsorption capacity for steroid compounds, meanwhile, to screen out and prepare the aptamers with exclusive recognition ability for steroid hormones by using SELEX technique. Then the obtained molecularly imprinted polymers and aptamers are used to construct an array enrichment unit and a recognition unit. Combined with graphene-nano-gold signal amplification unit, this project provides an on-line analytical method with a high sensitivity, strong specificity and strong anti-jamming ability by using the constructed electrochemical sensors. After validation and evaluation, the method could be applied to analyze the practical waste water in the production process. Coupled chemical recognition with biochemical identification, this proposal may provide a new idea and potential methods for the analysis of strong biological activity compounds during the drug production process.

甾体激素具有很强的生物活性，生产过程废液排放对生态构成严重威胁，目前对于此类物质的分析与监控尚处于起步阶段。本项目拟在前期工作基础上，针对甾体激素类药物生产过程中污染物分析方法专属性差、灵敏度低、分析过程滞后、分析对象与生理活性相关性差等问题，以典型的甾体激素为模板，采用印迹技术制备具有高容量吸附作用的分子印迹聚合物（MIPs）；根据甾体激素与细胞核中DNA靶点作用的特点，采用SELEX技术，筛选与制备具有专属识别能力的DNA适配体；利用MIPs和DNA适配体构建传感器的化学富集和生物识别阵列单元，采用石墨烯-纳米金材料构建传感器的信号放大单元；建立专属性强、灵敏度高、抗干扰能力强的甾体药物生产过程在线电化学分析方法，对分析方法进行验证与评价，并用于生产过程废液的在线分析。研究集甾体激素的化学识别与生物识别于一体，实现在线分析，为强生物活性药物生产过程的在线监控提供新的思路和方法。

甾体激素具有很强的生物活性，生产过程废液排放对生态构成严重威胁，目前对于此类物质的分析与监控尚处于起步阶段。针对甾体激素类药物生产过程中污染物分析方法专属性差、灵敏度低、分析过程滞后的问题，研究中一方面将分子印迹技术、材料表面修饰技术、固相萃取技术和液相色谱分析方法相结合，制备了一系列具备识别能力和快速分离能力的分子印迹聚合物（MIP）及复合纳米材料，将它们用作固相萃取吸附剂，建立了甾体激素的液相色谱分析方法，实现了对制药废水中微量甾体激素的特异性富集与检测。另一方面以导电性能优良的碳材料和金属等为信号增敏元件，以MIP和聚合氨基酸等为特异性识别元件，采用电聚合、电沉积或滴涂等手段将他们逐层修饰在电极表面，制备得到了一系列具有识别特性的电化学传感器，建立了甾体激素的电化学传感分析方法，并将其成功应用于制药废水中微量甾体激素的快速分析与灵敏检测。这两种分析方法的开发为甾体激素类药物生产过程中废液的在线监控和分析提供了新的思路。