This work describes the preparation and the physicochemical properties of atovaquoneloaded liposomes. It also describes drug release from the liposomes. As many factors can influence liposome stability, we studied several formulations, including different concentrations of atovaquone, phospholipids, and cholesterol. The effect of atovaquone (ATV) concentration was also evaluated. The highest binding percentage (100 ยฑ 2.5%) was obtained under alkaline conditions for a 2 mg/ml concentration of ATV. The percentage of encapsulation decreased significantly when drug concentrations increased. Drug uptake (expressed per unit mass of phospholipids) was nonlinearly related to equilibrium ATV concentration. A Langmuir-type sorption was suggested (r = 0.978). In acidic or neutral buffer, the binding percentage reached 42.1 ยฑ 0.02%. The increase of phospholipids and cholesterol concentrations did not significantly improve the ATV binding yield for the lowest ATV concentration. Conversely, ATV binding was significantly increased for the highest ATV concentration. All the formulations tested gave monodispersed liposomes with a mean diameter around 260 nm. The dilution (1/5-1/20) of liposomes in alkaline conditions induced a significant release of ATV (74% release). In acidic or neutral buffer no release was observed, suggesting an encapsulation process.