A novel hybrid material, which is an enzyme/inorganic nanosheet complex coated by molecularly imprinted polymer (MIP), was developed, and applied to colorimetric quercetin assay. First, an enzyme/inorganic nanosheet complex was prepared from horseradish peroxidase (HRP) enzyme and titanate nanosheet (TiOx), using electrostatic interactions between them in acetate buffer. In the next place, dopamine self-polymerization was performed in the presence of HRP/TiOx complex with quercetin as a template, to prepare MIP membrane onto the HRP/TiOx complex. After washing process, a new hybrid material, MIP-coated HRP/TiOx complex (MIP-HT) was obtained. MIP-HT adsorbed quercetin efficiently, compared with NIP-HT that is an HRP/TiOx complex coated with non-imprinted polydopamine. MIP-HT showed enzymatic activity for an oxidation reaction of guaiacol, which is a chromogenic substrate of HRP, whereas the enzymatic activity of NIP-HT was significantly suppressed. The amount of brown product, formed by the color reaction, reduced owing to the presence of quercetin in sample solution, and a good liner relationship was observed between the concentration of quercetin and the increment of absorbance at 470 nm. The investigation using several biomolecules indicates that MIP-HT has the ability to detect quercetin and its analogues with selectivity. Therefore, MIP-HT shows great promise as a new and attractive material for use in colorimetric assay of quercetin or quercetin analogues.
Keywords: Colorimetric assay; Horseradish peroxidase; Molecularly imprinted polymer; Polydopamine; Quercetin; Titanate nanosheet.
© 2022. The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry.