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Abstract: In accordance with the concept of Surface Imprinting, as proposed by the present authors, a Cu(II)-imprinted microsphere was studied in detail in order to obtain information about the origin of the imprinting effect. Unimprinted and Cu(II)-imprinted microspheres were prepared under several pH (4.0, 5.0, 5.6, or 6.0) conditions in second-step polymerization. The obtained microspheres were characterized regarding particle form, size-distribution pattern, average diameter, Cu(II) adsorption behavior, and so on. During preparation at pH 4.0-5.6 in the second-step polymerization, imprinted microspheres adsorbed Cu(II) more effectively than did unimprinted microspheres in this pH range, indicating a clear metal- imprinting effect. On the other hand, the imprinting effect was not observed during preparation at pH 6.0, because of the formation of copper(II) hydroxide precipitates. The maximum imprinting effect was provided by microspheres obtained at pH 5.6. Spectroscopic studies were made on Cu(II)-loaded microspheres. The FT-IR spectra suggested that carboxyl groups on the microsphere surface participated in the Cu(II) binding in their ionized carboxylate forms. An FE-SEM (angstrom SEM) observation of the microsphere indicated a smooth surface structure at the 10 nm level. The collected data indicate that the origin of the imprinting effect on a Cu(II)-imprinted microsphere is the interaction between Cu(II) and the carboxylate group at the aqueous-organic interface, supporting the concept of Surface Imprinting