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Background: The fluconazole transdermal delivery system as an antifungal provided to avoid side effects of nausea, dyspepsia and abdominal cramps. Transdermal delivery Fluconazole was made through the microemulsions system with tween 80:1-butanol as surfactant:co-surfactant. Objective: This study aimed to determine the physical stability of fluconazole microemulsions with increased concentration in tween 80 as surfactant and 1-butanol as cosurfactants. Materials and Methods: This study was initiated by determining the microemulsion with Isopropyl myristate as an oil phase where the comparison of tween 80:1-butanol 3:1. A clear area of ​​microemulsion was found in surfactant + cosurfactant concentrations of 22%, 23%, 24% and 25%. Based on this, 4 formulas for fluconazole microemulsions were made at surfactant concentrations:cosurfactant F1 (16.5%:5.5%) F2 (17.25%:5.75%) F3 (18%: 6%) F4 (18.75%: 6.25%). Further evaluation of physical stability included organoleptic, pH, cycling test, centrifugation, viscosity, particle size, zeta potential, and polydispersity index. Results: This study found the separation of phases in the formula 1 and 2, increased of pH of all the formula in every week of testing, and decreased of viscosity of formula 2 at week 4 while the other formulas experienced an increased. Whereas in PSA measurements showed particle size ranges from 20.2 to 90.1; polydispersity index 0.167-0.571; and zeta potential -6.02 to -20.3 mV. Conclusion: The increased concentration in tween 80 and 1-butanol affected the physical stability of microemulsions.