In this study, we report the development of adsorptive extraction materials by surface protein‐imprinted polymers (MIPs) over silica gel for selective recognition/separation of human serum albumin (HSA) from urine. The HSA‐imprinted polymers prepared on silica particle had at interface between the silica gel and different MIPs greatly produced enrichment for the binding of protein from the urine. The solid‐phase extraction of the optimized polymer layer was prepared by copolymerization of methacrylic acid (MAA), acrylamide (AAm), and dimethylaminoethylmethacrylate (DMAEMA) and a crosslinker methylenebisacrylamide (MBA) at the mole ratio of 1:158:88 (T:M:C) and showed moderate affinity (<104 order M−1) toward target protein HSA and selectivity. Four analogues, egg white albumin (EWA), bovine serum albumin (BSA), lysozyme (Lyz), and creatinine (Cre) were selected to study the binding efficiency of MIPs in single and binary protein solutions. We studied the influence on recognition ability for HSA and found that prepolymer mixture and matrix flexibility of the optimized thin polymer layer (35 ± 10 nm) on the submicrosilica particles. The high‐binding affinity (QMIP, 86.7 mg g−1) and fast kinetics (180 min) were observed for this synthesized HSA‐MIP when compared with other reported HSA‐MIPs in surface imprinting (5.9 and 11.3 mg g−1) and epitope surface imprinting (46.6 mg g−1) methods. We demonstrated the application in real and synthetic urine samples that the approach allowed the efficient adsorption of HSA in real urine (129.48 mg g−1) is almost double to the binding of HSA in synthetic urine (67.84 mg g−1). Apart from this, only minor interference of Cre (2.74 mg g−1) was observed, eventhough Cre is the final metabolite in urine. These adsorptive submicrosilica materials have potential in the pharmaceutical industry and clinical analysis applications. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 46894.

In this study, we have developed a method to assess adenosine 5ʹ‐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 (QMIP, 42.65 µmol/g), and high selectivity and specificity to adenosine 5′‐triphosphate compared to other competitive nucleotides including adenosine 5ʹ‐diphosphate, adenosine 5ʹ‐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 (QMIP, 100.3 µmol/g) and 2.51 for synthetic urine (QMIP, 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.