基于印迹纳米杂化膜及增敏探针双识别作用的SPR-SERS农药多残留研究

Rapid screening for multi-residues is the main developing direction in current rapid detection technology, and also is one of main technical bottlenecks of fast detection for pesticides in agro-products . The SPR and SERS can achieve the qualitative and quantitative analysis for pesticides residues, but they can not meet the need of the simultaneous determination for weak signal multi-pesticides in agricultural products due to poor signal stability, matrix interference and so on. In order to achieve the rapid and accurate determination of multi-pesticides residues in complex matrixes, this work is proposed to establish a highly sensitively, simultaneously quantitative-qualitative two-mode detection system of SPR specific recognition and SERS signal amplification based on the mechanism of surface plasmon resonance signal enhancement and the theory of competitive recognition of structure analogues via the research of signal amplification technology of multi-compounds labeled probe and efficient recognition technology of molecularly imprinted nano hybrid array membrane, by using two online modes and molecular competitive strategy of sample loading and elution in SPR. Based on that, this work is aimed at synthesizing ‘object - loaded gold carrier protein - labeled probe’ signal enhancement complexes of both SPR and SERS, constructing the SPR chip of molecularly imprinted nano hybrid array and the SERS substrate of multiple nano composite sensitization, studying each modified layer material to the influence of the probe's sensitivity and response, illustrating the mechanism of SPR multi-channel high bionic competitive recognition and control mechanism of interference factors, establishing the fast, efficient, qualitative-quantitative and simultaneous detection system of SPR and SERS, developing new methods for rapid detection of pesticide residues in agricultural products.

多残留快速筛查是当前速测技术的主要发展方向，也是农药快速检测面临的技术难点。表面等离子共振技术（SPR和SERS）可实现农药残留定性与定量分析，但存在信号稳定性差、基质干扰大等问题，无法满足农产品中信号特征较弱的多组分农药同步检测。本项目拟基于表面等离子共振信号增强机理与结构类似物竞争识别理论，构筑能同时提高SPR和SER两者信号增强的“多组分目标物-负载金载体蛋白-标记探针”，制备对多组分探针具有高灵敏响应的多元纳米复合增敏SERS基底，探明多组分信号探针分子的信号增强机制；合成具有高效识别作用的分子印迹纳米杂化阵列SPR芯片，研究探针分子与目标物对SPR分子印迹阵列膜的竞争吸附与解离规律，阐明SPR多通道高效仿生识别机理与干扰因素可控机制，构建SPR特异性识别与SERS信号放大的高灵敏同步定性定量双模式检测体系，拓展农药多残留实时在线快速检测技术的新方法和新途径。

多残留快速筛查是当前速测技术的主要发展方向，也是农药快速检测面临的技术难点。表面等离子共振技术（SPR和SERS）可实现农药残留定性与定量分析，但存在信号稳定性差、基质干扰大等问题，无法满足农产品中信号特征较弱的多组分农药同步检测。本项目拟基于表面等离子共振信号增强机理与结构类似物竞争识别理论，通过研究多组分标记探针信号多重放大技术与分子印迹纳米杂化阵列膜高效识别技术机理，构建 SPR特异性识别与 SERS 信号放大的高灵敏同步定性定量双模式检测体系，拓展农药多残留快速检测技术的新方法和新途径。.本项目通过EC-SPR制备了基于石墨烯/纳米金基底的分子印迹杂化膜，筛选和优化敏感仿生识别膜的聚合体系和杂化比例等制备条件，构建了最佳分子印迹纳米杂化膜制备工艺体系，利用EC-SPR联用技术表征杂化膜芯片掺杂去掺杂过的界面反应、吸附解离反应，获得具有高特异性识别草甘膦的分子印迹纳米杂化SPR芯片，并建立了其检测方法；设计合成一种类特异性识别含硫三嗪类农药的MIPs，制备了具有良好稳定性和高增强性能的纳米金SERS增强基底，研究了QuEChERS和MIPs两种前处理技术对SERS信号的影响机制，筛选出QuEChERS串联MIPs高效净化前处理技术，建立了小麦和水稻基质中扑草净和西草净残留检测的SERS方法；合成了Pt NPs@BSA-三唑磷半抗原复合模拟酶探针，探讨了模拟酶探针竞争响应机制，制备了Au NPs@MIL-101增强基底，创建了三唑磷农药模拟酶仿生比色/SERS快速检测方法。采用了原位还原法合成了Au NPs@MIL-101 SERS基底，合成了Pt NPs@BSA-三唑磷半抗原复合模拟酶探针，建立了分子印迹与模拟酶复合探针双重选择方法，成功应用于水及梨样品中三唑磷的检测；合成了Au NPs/4, 4-联吡啶/BSA-靶标半抗原复合标记探针，研制了分子印迹SPR芯片，筛选和优化了SPR-SERS联用的最佳缓冲体系和载样体系，建立了SPR-SERS联用技术。.课题取得了显著的成果，其中授权发明专利3个，并实现了成果转让，其转让金额1200万元，部分内容获国家技术发明奖1项，中国专利优秀奖1个，中国农科院成果转化奖1个，发表SCI论文36篇，培养研究生5人。