Flucytosine is chiefly employed in the antifungal eye drops formulation, especially recommended for treatment of serious infections. The poor bioavailability of eye drops failed to produce the maximum therapeutic effects to the patient. Hence, it was intended to prepare and evaluated in situ ophthalmic gel of Flucytosine for better therapeutic efficacy. The different proportion of the Xanthan gum and HPMC K4M along with sodium alginate were used for the preparation of in situ ophthalmic gel of Flucytosine. The six in situ ophthalmic gels were prepared, and evaluated for clarity, visual appearance, pH, gelling capacity, drug content, assessment of drug release, kinetic modelling, ocular irritancy and stability study. The formulation demonstrated appreciable findings for all the assessment. The F2 revealed greatest drug release for 8 hrs, and it implies the sustained release compared to other formulations. Further, the F2 followed the zero order drug release. The findings concluded that in situ ophthalmic gel of Flucytosine produce better therapeutic efficacy, and also reduce the frequency instillation into eye.