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

首先，分别以4-VPY和MAA为功能单体，通过多步溶胀-热聚合法制备了2-芳基丙酸衍生物和p-阻滞剂的分子印迹聚合物。然后，用GMMA和GDMA的1：1混合物进行亲水性表面修饰，并在分子印迹开始后4h直接添加到MIP中。结果表明，亲水性表面修饰后，目标药物的分子识别位保持不变，亲水性表面修饰后，牛血清白蛋白几乎完全从RAM-MIP中回收。上述结果表明，2-芳基丙酸衍生物或P-阻滞剂的MIP被选择性地修饰在亲水性的外层，利用RAM-MIP可以实现药物的直接血清注射分析。2-芳基丙酸衍生物或β-受体阻滞剂的RAM-MIP用于2-芳基丙酸衍生物或β-阻滞剂的直接血清注射分析。然而，印迹分子的泄漏阻碍了对药物的准确和精确的分析。通过将RAM-MIP用于结构上相关的化合物，这个问题已经被克服。该方法可广泛应用于生物体液中药物的测定。

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