QIP was prepared using quercetin as template, 4-VP as functional monomer and EDMA as the cross-linking monomer (Fig. 1). The choice of functional monomer is crucial for maintaining the stability of template–monomer complexes during the imprinting processes as well as pertinent in stabilizing the binding site and affinity. 4-VP was selected as a functional monomer owing to the H-bond acceptor property from the pyridine nitrogen. Such functional moiety of 4-VP is complementary to quercetin, which contained 5H-donor sites from hydroxyl groups. Previously, systemic investigation on the influence of monomer and crosslinking monomer on the properties of MIPs for quercetin had been carried out [33]. The suitable ratio of quercetin:4-VP:EDMA that afforded high specificity of MIPs for quercetin was 1:4:20, respectively. Meanwhile, MAA-containing polymer showed lower binding affinity for the template. Therefore, MIPs in this study were prepared using the molar ratio of 1:4:50. The binding performance of QIP was examined by varying the amount of polymers using a fixed amount of template. The binding analysis was performed in the same solvent used during polymerization (EtOH). Using 80 mg of polymer incubated against 0.1 mg/mL quercetin, the result is shown in Fig. 2. It can be observed that the QIP can bind to quercetin (44.8%) significantly greater than that of the control polymer (14.8%) by 3 folds of magnitude. To further enhance the binding performance of QIP, a small amount of H2O was added to the rebinding solvent as follows: ethanol:H2O(3:2) and ethanol:H2O(4:1) in order to promote the interaction via π–π stacking in the polar binary solvent mixture. Such findings were in accordance with the results from our previous reports [34] and [35] in which the binding performance of QIP and its control polymer in solvent mixture of EtOH:H2O (4:1,v/v) afforded good rebinding selectivity toward quercetin where MIP afforded 53.9% binding to its template molecule, which is 4.5 folds higher than that of the control polymer (12.5%) at 80 mg of polymer. The calculated binding capacity of QIP was approximately 1 mg/g of polymer. For rebinding analysis in ethanol:H2O(3:2) there were significant increases in non-specific binding as represented for 80 mg of both QIP and control polymer in which binding to the template was 69.6% and 46.2% respectively. Furthermore, the binding specificity of QIP was tested against tocopherol succinate, tocopherol nicotinate and 8-hydroxyquinoline. To 40 mg of QIP, each of various analyses (0.1 mg/mL) was added. QIP could bind quercetin up to 40% whereas it could bind tocopherol succinate, tocopherol nicotinate and 8-hydroxyquinoline at less than 5%. Results indicated that QIP displayed high selective recognition toward its template than those of the other analytes as shown in Fig. 3.