The use of in situ gels as an eye drug delivery mechanism has been thoroughly investigated to increase bioavailability and efficacy. The purpose of the current study was to use an animal model to assess the effectiveness of prepared in situ gel. In vivo irritation and pharmacokinetics of the improved formulation (G7) were investigated in rabbits. All the physicochemical characteristics of G7 were within acceptable bounds. The results of the investigation on ocular irritation show that the chosen formulation is secure and non-irritating when administered intravenously. As shown by higher Cmax Cmax (834±47 μg/ml) and AUC (2451± 94 μg h/ml) when compared with commercial eye drops (Cmax: 503±38 μg/ml and AUC: 876±54 μg h/ml), in-vivo pharmacokinetics data shows a substantial enhancement of fluconazole bioavailability (p 0.0001) from G7.