Development of sensitive and selective method for the determination of drugs and their metabolites in biological fluids using artificial antibodies

首先，以4-VPY和MAA为功能单体，通过多步溶胀和热聚合法制备了2-芳基丙酸衍生物和p-阻滞剂的mip；接下来，用1:1的GMMA和GDMA的混合物进行亲水性表面修饰，在分子印迹开始4小时后直接加入药物的MIP中。结果表明，经过亲水表面修饰后，靶药物的分子识别位点保持不变。此外，在对RAM-MIP进行亲水性表面修饰后，牛血清白蛋白几乎完全从RAM-MIP中恢复。上述结果表明，2-芳基丙酸衍生物或p受体阻滞剂的MIP是通过亲水性外层选择性修饰的，使用该RAM-MIP可以实现药物的直接血清注射测定。所获得的2芳基丙酸衍生物或p受体阻滞剂的RAM-MIP通过柱切换系统（由RAM-MIP和常规CI S-silica柱组成）用于药物的血清直接注射测定。然而，印迹分子的泄漏阻碍了对药物进行准确和精确的分析。通过使用RAM-MIP来制备结构相关的化合物，克服了这一问题。所开发的方法应广泛适用于生物液体中药物的测定

First, the MIPs for 2-arylpropionic acid derivative and p-blocker were prepared using 4-VPY and MAA, respectively, as functional monomers by a multi-step swelling and thermal polymerization method. Next, a 1 : 1 mixture of GMMA and GDMA was used for hydrophilic surface modification, and it was added directly to the MIP for the drug 4 h after the start of molecular imprinting. The results revealed that the molecular recognition sites of the target drug remained unchanged with hydrophilic surface modification.Further, after hydrophilic surface modification of the MIP, bovine serum albumin was almost completely recovered from the RAM-MIP. The results described above reveal that the MIP for 2-arylpropionic acid derivative or P-blocker is selectively modified with hydrophilic external layer, and that direct serum injection assays of the drug could be attained using the RAM-MIP. The obtained RAM-MIP for 2arylpropionic acid derivative or p-blocker was applied for direct serum injection assays of the drug by column-switching system, consisting of a RAM-MIP and a conventional CI S-silica column. However, leakage of the imprint molecule prevented accurate and precise assays of the drug. This problem has been overcome by using the RAM-MIP for a structurally related compound. The developed method should have wide applicability for the determination of a drug in biological fluids