典型甘油醚类化合物及其衍生物的平面波导荧光免疫分析方法构建与环境行为研究

Bisphenol A diglycidyl ether (BADGE), novolac glycidyl ether (NOGE) and their derivatives are typical glyceryl ethers that are widely used in life and industrial productions. Studies reported endocrine-disrupting, genotoxic and cytotoxic potentials of the containments, and the chemicals could be discharged to environments and result in serious environmental problems. However, up to date, the environmental distribution and ecological risk of the pollutants are not clear, which is mainly due to lack of the suitable analytical methods. Currently the traditional instrument approaches suffer from complicated procedures in sample pretreatment, and time consuming to pre-concentrate and separate targets from matrices, as a consequence, the systematic results can not be obtained after measurement of massive environmental samples within a short time. Therefore, it is necessary to develop a simple and rapid immunoassay to monitor the organic containments. In the project we will design and synthetize a series of haptens, based on this, monoclonal antibodies against these compounds can be produced, after labelled by fluorescent reagent, a quick, simple, sensitive planar waveguide fluorescence biosensing system with good stability and high tolerance to numerous matrix interferents could be established for analysis of these chemicals from a large number of different environmental, biological and human samples in some typical regions. The distribution, transportation and bioaccumulation of these chemicals in contaminated regions will be studied in detail, our results will provide a scientific basis for further in-depth understanding of the environmental and health risks of these pollutants.

双酚A二缩水甘油醚(BADGE)、酚醛清漆甘油醚(NOGE)及其衍生物是一类在生活与工业生产中广泛使用的典型甘油醚类化合物，具有内分泌干扰效应、细胞与基因毒性。这类物质被排放到环境中，将导致严重的环境污染问题。然而，目前其污染特征与环境风险仍不明确，方法学是制约这一问题的关键因素。现有的仪器分析法（SPE-LC/MS/MS）因前处理繁琐、分析时间长等不足，无法在较短时间内通过大量样本的检测得出规律性的结论。因此，发展简单快速的免疫分析方法极其必要。本项目拟分别制备可识别BADGE、NOGE及其衍生物的单克隆抗体，再以荧光试剂标记，构建前处理简单、灵敏度高、稳定性好的平面波导荧光免疫传感器，对污染物进行快速分析；通过高频次、多样品、长期的区域采样与分析，系统研究这些化合物在典型区域环境中的分布、迁移及生物富集过程，为进一步深入了解此类污染物的环境暴露风险并制定有关防控对策提供科学依据。

针对BADGE及其衍生物，设计并合成了一系列半抗原，采用碳二亚胺法和活泼酯法将半抗原与大分子蛋白偶联制备免疫抗原和包被抗原，并通过免疫新西兰大白兔得到八种抗血清。基于此构建了针对此类污染物的间接竞争免疫分析方法，在此基础上，通过引入纳米材料，及提高酶载量的方式进行信号放大，建立了一系列高通量、高灵敏、快速生物分析方法，部分方法学探索结果，将有较好的应用前景。利用所构建的生物分析方法，对典型区域及罐头食品中此类化合物的污染水平与现状进行了系统的调查研究，结果表明，罐头食品中普遍检出了此种污染物，但最高浓度均在1ng/g之下，且BADGE及其衍生物的总量随着时间和温度的增加呈现递增的趋势。但在典型区域采集的多介质环境样品与生物样本检测的结果表明，其检出率较低，含量极低，对生态环境与人体健康未造成负面影响。