To assess whether there are variations in the intraocular pressure (IOP) between the morning and afternoon in healthy subjects.This study was carried out in a Primary Care Health Center among usual patients of general practitioners. Two measurements of the intraocular pressure were assessed on the same day (between 8 and 9 a.m. and between 5 and 6 p.m.) using a Perkins applanation tonometer. The subjects (119 females and 101 males) had a mean age of 46.3 years, with ages ranging between 14 and 83 years. They were proportionally distributed among 4 age groups (14-24, 25-44, 45-64 and >65 years).The IOP mean values were: right eye, morning time and afternoon, 13.29 and 11.81 mm Hg, respectively (p<0.001); left eye, morning time and afternoon, 13.48 and 12.04 mm Hg (p<0.001). The IOP was significantly higher in males than in females, and showed a weak association with age.We found a diurnal variation of IOP of approximately 1.5 mm Hg. The time of day when the IOP is measured needs to be considered as a variable in research in ocular hypertension and glaucoma.