识别氨基糖苷类抗生素的比率型荧光印迹阵列传感基础研究

Compared with traditional rapid detection techniques, fluorescent imprinted array sensor exhibits unique superiority in analysis of multi-targets. However, the construction of fluorescent imprinted array sensor still remains a great challenge due to limited number of fluorescent monomers and competitive adsorption indicator. In this project, a new fluorescent response strategy for imprinted recognition of aminoglycosides based on bi-recognition competitive adsorbent for imprinted sites and ratiometric fluorescent probe is proposed for the first time. As a result, a novel ratiometric fluorescent imprinted array sensor will be developed. Additionally, the response rule of the fluorescent system will be comprehensively investigated. Furthermore, a new method for rapid detection of multiple aminoglycosides in complex food samples will be achieved. In summary, the project integrates the advantages of specific imprinted recognition, ratiometric fluorescent probe, and array analysis for detection of veterinary drugs, providing a new research platform for developing rapid detection technique.

荧光印迹阵列传感技术较传统快速检测技术在多靶标分析中显示出独特的优势，但是由于荧光单体或竞争吸附指示剂难寻，荧光印迹阵列传感器的构建仍然存在巨大挑战。本项目拟以氨基糖苷类抗生素为研究对象，首次提出基于印迹位点-比率型荧光探针双识别竞争吸附剂的传感新策略，在此基础上，发展一类比率型荧光印迹阵列传感器；探明传感体系对氨基糖苷类抗生素的响应规律，并开发复杂食品中多种氨基糖苷类抗生素残留同步检测的新方法。本项目研究的意义在于原创性地将分子印迹特异性识别、比率型荧光检测、阵列高通量分析三大优势相结合用于兽药残留分析领域，为开发快速检测技术开辟新的研究思路。

荧光印迹阵列传感技术具备高灵敏度荧光检测、高通量传感、分子印迹选择性识别三大技术优势，其在食品分析领域的应用前景可谓一片蓝海。然而，目前印迹荧光单体及竞争吸附荧光指示剂种类十分有限，加之需要大量繁琐的考察筛选实验才能获得，这为荧光印迹阵列传感器的构建带来巨大挑战。本项目以氨基糖苷类抗生素等水溶性风险因子为研究对象，探索了鸡肉等复杂食品基质的样品净化材料及净化方法，发展了基于印迹位点-比率型荧光探针双识别竞争吸附剂的荧光阵列传感器，并在此基础上，构建了典型氨基糖苷类抗生素的快速检测方法。主要研究内容及进展包括：（1）创制了聚丙烯酰胺基葡萄糖亲水性磁性固相萃取吸附剂、两亲性嵌段共聚物磁性QuEChERS净化吸附剂，高效去除蛋白质、色素、长链脂肪酸、小分子有机酸等食品基质干扰物，并且实现快速磁分离，单个样品净化时间不超过30 s，通过亲水相互作用模式实现对链霉素、卡娜霉素、大观霉素等10种氨基糖苷类抗生素同步富集，利用液相色谱串联质谱技术，检测鸡肉中10种氨基糖苷类抗生素，检测方法的检出限为0.6 μg kg-1-23.6 μg kg-1。（2）制备了氨基糖苷抗生素MIP材料、双识别竞争吸附剂、polymer-PyBr/-CD比率荧光探针，并进一步构建MIP比率荧光传感器，初步实现对链霉素的选择性筛查识别，荧光响应时间为5 min，检测线性范围50 ng mL-1-500 ng mL-1。本研究为发展具有自主知识产权的原创性分子印迹荧光阵列传感器提供了一条新的思路。