Glasses and silica-gel packings modified with fluorocarbon and hydrocarbon silylation agents were prepared to investigate the separation mechanism of a fluorocarbon bonded layer in reversed-phase high-performance liquid chromatography (RP-HPLC). On the hydrocarbon layer the contact angles of benzene (Be) and hexafluorobenzene (FB) decreased with increasing carbon chain length of the modifier, and the mobile phase (MP) was independent of the modifier. The fluorocarbon layer gave a larger contact angle for each liquid than the hydrocarbon layer. The retention factor, k, of Be and FB on hydrocarbon packings increased with decreasing contact angle. It was demonstrated that the separation of Be and FB on hydrocarbon packings is caused by solvophobic interaction between solute molecules and the stationary layer. In contrast, k increased with increasing contact angle at the fluorocarbon layer. The retention of Be and FB on fluorocarbon packings may be attributed to the penetration of the solute molecules from the mobile phase into the fluorocarbon stationary layer.