Objectives The aim of this study was to evaluate the effect of topical tropicamide at two concentrations (0.5% and 1.0%) on the Schirmer tear test (STT) results in clinically normal cats. Methods Twenty-four adult domestic shorthair cats were randomly assigned to three groups. In all three groups, ophthalmic solutions were instilled in a randomly selected eye and the opposite eye served as the control. In groups 1, 2 and 3 one drop of 0.5% tropicamide, 1.0% tropicamide and distilled water was used, respectively. Tear production in both eyes was tested 30 and 60 mins after instillation in all three groups. Results Baseline mean +/- SEM STT values for the treated eyes in groups 1, 2 and 3 were 13.37 +/- 2.91 mm/min, 10.87 +/- 1.39 mm/min and 11.37 +/- 1.65 mm/min, respectively. Thirty minutes after the drug instillation in the treated eye, mean +/- SEM STT values in groups 1, 2 and 3 were 4.87 +/- 3.05 mm/min, 2.00 +/- 0.84 mm/min and 11.25 +/- 1.81 mm/min, respectively. The mean +/- SEM STT levels of the treated eye after 60 mins were 3.75 +/- 1.87 mm/min, 0.5 +/- 0.37 mm/min and 11.42 +/- 1.78 mm/min in groups 1, 2 and 3, respectively. Conclusions and relevance Use of 0.5% topical tropicamide, instead of 1.0% tropicamide, as a mydriatic agent, can be recommended in cats. Tear production measurement should be performed prior to the administration of tropicamide, regardless of the concentration of this drug.