基于限进性分子印迹荧光传感的血清中甲氨蝶呤快速分析方法研究

Methotrexate is widely used in the treatment of a number of cancers, and the safety of its application has draw much attention in clinical. Particularly, the proper usage of calcium folinate can reduce the side-effect and improve the safety of methotrexate therapy. However, The perfect rescue of calcium folinate depends on the monitoring of the concentration of methotrexate in blood rapid and accurate. At present, the methods for methotrexate detecting require tedious derivations and pretreatments, which make the analysis time-consuming and lack of selectivity. In this study, the self assemble of the template molecule and functional monomer is used to directly synthesize a restricted molecularly imprinted polymers on the surface of quantum dots with the addition of water-compatible cross-linker. The arrangement of the binding sites constitutes an induced molecular memory, which makes the prepared polymers capable of recognizing the target. The prepared restricted-accessed molecularly imprinted fluorescent polymers can be used for the selective recognition and fast fluorescence detection for methotrexate in the complex biological sample. Thus, the method has advantages of rapid and convenient operation, high fluorescence efficiency, unnecessary derivation and shows excellent selectivity in serum. Subsequently, the mechanism of molecular recognition and luminescent response of the polymers are investigated. According to the optimization results, the proposed method can be adopted for the detecting of methotrexate in serum, which has an excellent perspective of practical applications in the future. This technology resolves the problems of traditional method for methotrexate detecting, improves the development of molecularly imprinted polymers in the field of clinical treatment and provides a new method for the development of precision medicine.

甲氨蝶呤是一种临床广泛使用的抗肿瘤药物，其治疗安全性是临床关注的焦点。快速而准确的监测患者血清中甲氨蝶呤的药物浓度，使用其特殊解救剂亚叶酸钙解救可减少治疗副作用、提高用药安全性。目前甲氨蝶呤的检测需要繁琐的衍生试剂衍生和提取前处理，操作费时且缺乏选择性，难以实现快速且准确的分析。本研究拟通过模板分子/功能单体的合理自组装，引入亲水交联剂，在量子点荧光材料的表面合成具有选择性识别特性的限进性分子印迹聚合物，简化样品前处理过程、降低血清中蛋白等成分的干扰，实现甲氨蝶呤的高选择性识别和快速荧光分析，并系统研究该聚合物对甲氨蝶呤的选择性识别及荧光响应机理。在此基础上，建立一种甲氨蝶呤的快速分析方法，对患者血清中的甲氨蝶呤药物浓度进行检测。本研究克服了甲氨蝶呤常规检测方法的不足，提供了一种甲氨蝶呤的快速准确分析方法，促进了分子印迹识别材料在临床治疗领域的应用和发展，为精准医疗的发展提供了一种新思路。

甲氨蝶呤是一种临床应用广泛的抗肿瘤药物，其治疗安全性是临床关注的焦点。快速而准确的监测患者血清中甲氨蝶呤的药物浓度，使用其特殊解救剂亚叶酸钙解救可减少治疗副作用、提高用药安全性。目前甲氨蝶呤的检测需要繁琐的衍生试剂衍生和提取前处理，操作费时且缺乏选择性，难以实现快速且准确的分析。在本课题中以建立新的、简单、快速、选择性高的甲氨蝶呤检测方法为首要目标，依次建立了四种甲氨蝶呤检测新方法，其中三种电化学检测方法，一种磁性固相萃取-高效液相色谱联用的检测方法。根据这四种方法的建立本课题研究共分为5个部分：1.基于乙炔黑修饰的玻碳电极快速检测血清中甲氨蝶呤2.十八烷基三甲基溴化铵修饰的乙炔黑糊电极检测血清中甲氨蝶呤3.甲氨蝶呤亲水性磁性分子印迹聚合物的制备与性能评价4.基于磁性分子印迹聚合物快速选择性检测血清中甲氨蝶呤5.基于甲氨蝶呤磁性分子印迹聚合物的磁性固相萃取-高效液相色谱联用方法检测试剂血样中甲氨蝶呤的研究。每部分都对各影响检测及制备的条件进行了详细的优化，得到了每部分方法建立及材料制备的最优条件，且对方法进行了验证，结果显示所建立的方法准确、快速、简便。本研究克服了甲氨蝶呤常规检测方法的不足，提供了新的甲氨蝶呤的快速准确分析方法，促进了分子印迹识别材料在临床治疗领域的应用和发展，为精准医疗的发展提供了一种新思路。