In this study, silica spheres (SiO2) nanomaterials were introduced as the support materials for glucose sensors. Glucose oxidase (GOx) was immobilized onto SiO2 spheres (SiO2/GOx) using physical adsorption (electrostatic interaction) method. The SiO2 spheres were synthesized in-house using a nonsurfactant template of palm oil derived fatty alcohols (PODFA). Based on FT-IR analyses, SiO2/GOx exhibited N-H absorbance at ca. 2990 cm-1 and O-H absorbance at ca. 3330 cm-1. One new absorbance peak was observed at ca. 2320 cm1 attributed to the bending vibration (νbend)of silane molecules (Si-H) obtained by the interaction of GOx and the surface silanols (Si-OH). UV-Vis analysis results exihibited the presence of a new broad peak for GOx/SiO2 sample at ca. 250 nm to 280 nm suggested to be assigned to H2O2, C=C and benzene ring from GOx. The optimum pH and optimum temperature for SiO2/GOx for GOx activity were ca. 7.0 and ca. 50 °C respectively. The increase of glucose concentration from 2mM to 5mM resulted to the increment of absorbance value for both GOx and SiO2/GOx suggesting succesful immobilization of GOx on SiO2 spheres. These materials were suitable for glucose detection at very small glucose concentrations particularly in salivary glucose detection.