Grafting silicon wafers with CH3O-(CH2-CH2-O)(n)-C3H6-trimethoxysilane and -trichlorosilane (n = 6 to 9) was performed in different conditions (solvent, reaction time, washing) in order to select procedures compatible with the design of nanostructured surfaces for biomaterial applications, using electron-beam lithography. After a first screening by principal component analysis (PCA), the X-ray photoelectron spectroscopy (XPS) data were analyzed by plotting the carbon to oxygen molar ratio vs the molar ratio of carbon singly bound to oxygen [C-O] over carbon bound only to carbon and hydrogen [C-(C,H)]. This was found to be a convenient method for discarding samples containing free polymerized silane. Such excess occurred as a result of insufficient washing or unsuitable solvent for the reaction (ether), as confirmed by AFM and thickness measured by X-ray reflectometry. Angle resolved XPS analysis indicated that the grafted silane layer had a 1-2 nm thickness and was covered by a thin layer of adventitious contaminant. As a result, the surface chemical composition obtained covered a broad range (O/C of 0.4 to 1.1; C-O/C-(C,H) of 2.5 to 6.5); variations could not be related to the nature of the silane reagent and no significant difference was found between hexane and toluene as solvent for the reaction. The grafted silane layer was not stable upon incubation during 24 h in phosphate buffered saline (PBS) at 37 degrees C, which mimics biological environments. As a consequence, the grafted wafers did not show protein repellent properties. This alteration was not observed at room temperature. XPS analysis demonstrated that silane layer detachment was due to a hydrolysis within the SiO2 layer initially present at the wafer surface.