Molecular Recognition Materials Research Unit, Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat Yai, Thailand.
J Sep Sci. 2019 Dec;42(24):3662-3678. doi: 10.1002/jssc.201900583. Epub 2019 Oct 22.
In this study, we have developed a method to assess adenosine 5'-triphosphate by adsorptive extraction using surface adenosine 5'-triphosphate-imprinted polymer over polystyrene nanoparticles (412 ± 16 nm) for selective recognition/separation from urine. Molecularly imprinted polymer was synthesized by emulsion copolymerization reaction using adenosine 5'-triphosphate as a template, functional monomers (methacrylic acid, N-isopropyl acrylamide, and dimethylamino ethylmethacrylate) and a crosslinker, methylenebisacrylamide. The binding capacities of imprinted and non-imprinted polymers were measured using high-performance liquid chromatography with UV detection with a detection limit of 1.6 ± 0.02 µM of adenosine 5'-triphosphate in the urine. High binding affinity (Q , 42.65 µmol/g), and high selectivity and specificity to adenosine 5'-triphosphate compared to other competitive nucleotides including adenosine 5'-diphosphate, adenosine 5'-monophosphate, and analogs such as adenosine, adenine, uridine, uric acid, and creatinine were observed. The imprinting efficiency of imprinted polymer is 2.11 for urine (Q , 100.3 µmol/g) and 2.51 for synthetic urine (Q , 48.5 µmol/g). The extraction protocol was successfully applied to the direct extraction of adenosine 5'-triphosphate from spiked human urine indicating that this synthesized molecularly imprinted polymer allowed adenosine 5'-triphosphate to be preconcentrated while simultaneously interfering compounds were removed from the matrix. These submicron imprinted polymers over nano polystyrene spheres have a potential in the pharmaceutical industries and clinical analysis applications.
在这项研究中，我们开发了一种通过使用表面腺苷 5'-三磷酸印迹聚合物在聚苯乙烯纳米粒子（412 ± 16nm）上进行吸附萃取来评估腺苷 5'-三磷酸的方法，用于从尿液中选择性识别/分离。印迹聚合物是通过乳液共聚反应合成的，以腺苷 5'-三磷酸为模板、功能单体（甲基丙烯酸、N-异丙基丙烯酰胺和二甲基氨基乙基甲基丙烯酸酯）和交联剂亚甲基双丙烯酰胺。使用高效液相色谱法（HPLC）结合紫外检测法，通过检测尿液中腺苷 5'-三磷酸的检测限为 1.6±0.02µM，测量了印迹和非印迹聚合物的结合能力。与其他竞争性核苷酸（包括腺苷 5'-二磷酸、腺苷 5'-单磷酸和类似物如腺苷、腺嘌呤、尿嘧啶、尿酸和肌酸）相比，印迹聚合物具有高的结合亲和力（Q，42.65µmol/g）、高选择性和特异性。与尿液（Q，100.3µmol/g）和合成尿液（Q，48.5µmol/g）相比，印迹聚合物的印迹效率分别为 2.11 和 2.51。该萃取方案成功应用于从加标人尿中直接提取腺苷 5'-三磷酸，表明这种合成的分子印迹聚合物允许在同时去除基质中干扰化合物的情况下，对腺苷 5'-三磷酸进行预浓缩。这些亚微米印迹聚合物在纳米聚苯乙烯球上具有在制药工业和临床分析应用中的潜力。
