The eye is a paired organ, the organ of vision. The eye is made up of various components, which enable it to receive light stimuli from the environment, and deliver these stimuli to the brain in the form of an electrical signal. The aim of this research was to develop formulation of the mucoadhesive nanoparticles of Moxifloxacin (BCS Class I drug) to increase residence time and bioavailability for the effective treatment of various ocular disorders like Glaucoma, conjunctivitis, corneal ulcer, trauma etc. The 12 trials batches with different ratios of excipients were taken, out of these three batches were blanks and nine were formulated with drug and best formulations were selected. The PDI of the formulations was varied from 0.137 to 0.512. Particle size of formulation varies from 183.7 nm to 962.1 nm. Zeta potential on the particles determines their physical stability (high zeta potential leads to more stable the colloid particles). The zeta potential of the formulations was varied from 14.2 to 27.5.Entrapment Efficiency of all formulation was found to be in the range of 39.80 to 94.24. The percent loading capacity of all formulation were found to be in the range of 3.32 to 7.87. Optimized formulation (F2)&marketedeyedropsofMoxifloxacinHClwereevaluatedforantimicrobial activity by cup-plate method. Formulation (F2) gave a clear zone of inhibition comparable with the zone of inhibition given by marketed eye drops. Results showed that formulation has better antimicrobial efficacy compared with the marketed eye drops.