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The aim of this work was to develop a freeze-dried microemulsion using Design of Experiment and loaded with Amphotericin B, which is quite unstable in aqueous media, as a drug model. Different types of cryoprotectants were studied. The microemulsion containing Maltose at 5% (w/w), frozen at the temperature of-80 °C, and performed with a 24h of freeze-drying time yielded the best results. The freeze-drying process reduced the microemulsion droplet size and don't change the AmB content. Therefore, microemulsions containing Maltose are suitable for drug incorporation and the freeze-drying was able to enhance the drug stability in the system.