The composition of ophthalmic preparation is administered topically to the eye in the form of a solution, suspension, ointment, gel or foam for the purpose of treating eye disease. Virgin Coconut Oil (VCO) has been one of the desired ingredients of choice, as its benefits as functional food oil is known among the public. The uniqueness of coconut oil is its fats and oils that contain the highest percentage of medium-chain fatty acids (MCFA), which have antimicrobial properties, such as lauric acid and capric acid. This study aimed to evaluate the antimicrobial effects of eye drop containing a different VCO concentration using the Kirby-Bauer test. The formulation of eye drop had 1.5%, 2% and 3% amount of virgin coconut oil, which was later added with all basic materials needed for eye drop. The samples were evaluated for its zone of inhibition (ZOI). The antimicrobial effects of eye drop formulation that contains 3% of VCO (F3) against Streptococcus pneumonia were similar with all control products (Eye Glo, Pred Forte, Cationorm), and antimicrobial effects of F3 against Staphylococcus aureus are better than Cationorm. It is noticeable that the higher the VCO content in the formulation, the better the antimicrobial effects of the eye drop. In conclusion, VCO possesses moisture, anti-inflammation, better anti-microbial properties, and it could be further formulated as a stable eye drop emulsion.