We examined the behavior of various entrapped guest molecules within silica hydrogel and evaluated the effect of Coulombic interactions and physical confinement on molecular mobility. Although rhodamine 6G (R6G) and fluorescein (FL) share similar size and molecular structure, their behavior in silica hydrogel was found to be dramatically different. A good majority of R6G was immobilized with little to no exchangeable molecules, whereas FL displayed a considerable amount of mobility in silica hydrogel. Moreover, silica hydrogel encapsulated R6G failed to gain mobility even under low pH or high ionic strength conditions to minimize Coulombic interactions, implying that encapsulated R6G molecules were inaccessible and likely trapped deep inside the silica matrix of a hydrogel. On the contrary, FL was relatively free to rotate and translate inside a silica hydrogel, implying that FL remained solvated in the solvent phase and was able to maintain its mobility throughout the hydrogel formation process. Fluorescence recovery after photobleaching measurements Put the diffusion coefficient of FL in silica hydrogel at ca. 2.1 x 10(-6) cm(2) s(-1), about a factor of 3 slower than that in solution. The substantial difference in mobility between cationic R6G and anionic FL led us to conclude that the effect of Coulombic interactions on mobility is more dominating in hydrogel than in alcogel. Our results also suggest that Coulombic interactions are strong enough to influence the eventual placement of a guest molecule in a silica hydrogel, causing R6G and FL to reside in different microenvironments. This has a profound implication on the use of molecular probes to study silica hydrogel since a slight difference in physical attribute may result in very diverse observations even from identically prepared silica hydrogel samples. As demonstrated, the repulsion between FL and silica renders FL liquid-bound, making FL more suitable for monitoring the change in viscosity and physical confinement during hydrogel formation, whereas other researchers have shown that silica-bound R6G is more suitably used as a reliable probe for monitoring the growth of silica colloids because of its strong attraction toward silica.