Surfaces that exhibit a gradual change in their chemical and/or physical properties are termed as surface gradients. Based on the changes in properties they are classified either as physical or chemical gradients. Chemical gradients show variations in properties like polarity, charge, functionality concentration and have found potential applications in fields of biology, physics, biosensing, catalysis and separation science. In this dissertation, surface gradients have been prepared using controlled rate infusion (CRI). CRI is a simple method in which a surface gradient is formed by carrying out the infusion of organoalkoxysilane in a time-dependent fashion using a set infusion rate. Depending on concentration of silane, rate of infusion and time of infusion, the gradient profiles on surfaces can be varied and the surface chemistry of the substrate can be altered. Initial work in the dissertation focuses on demonstrating different gradient profiles and selectivity obtained using amine and/ or phenyl functionalized gradient stationary phases on thin layer chromatography (TLC) plates prepared by CRI. The presence of amine and phenyl on the surfaces were confirmed by X-ray Photoelectron Spectroscopy (XPS) and diffuse reflectance spectroscopy, respectively. The change in surface chemistry was demonstrated by changes in the selectivities of water and fat soluble vitamins. After successful preparation and characterization of single and multi-component stationary phase gradients for planar chromatography, single-component gradients were prepared for column chromatography (Silica monolithic columns). Similar to that observed for planar chromatography, the selectivity was evaluated from retention factors and was found to be different for a weak acid/weak base mixture. The results obtained showed the promising approach of using gradient stationary phases in column chromatography. This work was further extended to prepare amine and phenyl multi-component gradients on silica monolithic columns to investigate mixed-mode and synergistic effects. Finally, amine, phenyl and thiol gradients were also prepared on cellulose substrates, particularly water color paper, The goal was to study the formation of functionality gradients on cellulose substrates particularly the interaction between hydroxyl groups on cellulose and silanols and to study the stability of the silanes on the cellulose surface.