Mesoporous MCM-41 silica spheres were prepared via the pseudomorphic route, using commercial prontosil silica spheres. Surface modification offers a great opportunity to adjust both the pore structure and surface properties of MCM-41 type materials which results in materials of improved hydrothermal and mechanical stability to make them promising candidates for chromatographic applications. In the present context, the obtained MCM-41 silica spheres were surface modified with octadecyl (C18) alkyl chains, by using direct grafting and surface polymerization methods, in order to vary the degree of surface hydrophobicity. The resulting materials were characterized before and after surface modification using various characterization techniques, with special emphasis on NMR and FTIR spectroscopies, for studying the attachment, mobility and the conformational order of the attached alkyl chains.