Microemulsions are isotropic mixtures of oil and water stabilized by surfactant and excellent drug delivery carriers due to their typical properties. Their antimicrobial activity can be due to direct interaction with fungal cytoplasmic membrane. The present study aims to enhance the solubility of hydrophobic drug using formulated novel microemulsion system and check probable synergistic impact of oil system along with conventional drug against fungal strain of C. albicans. The worked out optimized formulation was achieved through oil: surfactant: water (5:20:70) which was subjected to physiochemical analysis. The rate of drug release was checked for drug loaded microemulsion compared to conventional drug (Ketoconazole). The drug loaded and unloaded optimized microemulsion formulations were checked for its antimicrobial activity against C. albicans, for its minimum inhibitory concentration (MIC) along with cytoplasmic leakage using membrane integrity assay. Kill kinetics study was carried out to check the time-based lethality of Ketoconazole based microemulsion against the fungal strain. In vitro toxicity was analyzed against human lymphocyte for microemulsion. The appearance of morphological distortion was observed using fluorescent staining and SEM. The results indicate promising delivery vehicle for the intended purpose and the optimized emulsion formulation was found to be non-toxic at lower concentration for oral usage. © 2018 Elsevier B.V.