Four types of copper-containing MCM-41 mesoporous silicas were synthesized by the surface organometallic chemistry (SOMC) procedure (Cu/MCM-41-S), mechanical mixing (Cu/MCM-41-M), impregnation (Cu/MCM-41-I), and the hydrothermal technique (Cu/MCM-41-H). The resultant samples were characterized in detail by X-ray diffraction (XRD), N-2 physical adsorption, transmission electron microscopy (TEM), UV-vis diffuse reflectance spectroscopy (UV-vis DRS), temperature-programmed reduction (TPR), and infrared spectroscopy (IR) of NO adsorption. Catalytic behaviors of these samples for hydroxylation of phenol with H2O2 were evaluated. The results revealed that depending oil the preparation methods the samples contain different copper-oxo species and thus show different catalytic behaviors. Among these samples, the one prepared by SOMC contains a predominant amount of isolated Cu2+ and exhibits the most excellent catalytic activity and selectivity. The amount of isolated copper species decreases in the order of Cu/MCM-41-S > Cu/MCM-41-H > Cu/MCM-41-I > Cu/MCM-41-M, while the amount of copper oxide clusters increases in a reversal order. The difference in the catalytic activity and product selectivity of these four samples could be rationally explained by the distinction of chemical states of copper species. The highly dispersed isolated Cu2+ species are identified as the active sites in the phenol hydroxylation, while the nonisolated Cu2+ clusters or oxide are responsible for the deep oxidation of primary product HQ and the decrease of product selectivity. The mechanism of the copper-catalyzed phenol hydroxylation was proposed.