The purpose of this study was to evaluate the effect of timolol hemihydrate 0.5% (Betimol [THH], Ciba Vision Ophthalmics) and timolol maleate gel forming solution 0.5% (Timoptic-XE , [TXE], Merck, Inc.), when both are dosed once daily on the exercise performance. Maximum exercise heart rate reflects systemic beta-blockade activity. Fourteen healthy subjects were randomized to receive either placebo (HypoTears , Ciba Vision Ophthalmics), THH, or TXE by a Latin square technique in a three period crossover design. Subjects were dosed one drop every morning beginning three days before exercise testing. The interval between each test was one week. Exercise testing was performed two hours after dosing. Maximum exercise heart rate showed no statistical difference between TXE and THH (174 +/- 13.1 vs. 172 +/- 14.9 beats/min, respectively, P = 0.72). Both active treatments, however, decreased heart rate compared to placebo (185 +/- 7.3 beats/min, P = 0.017). Time to exhaustion showed no difference between groups (P > 0.10). The double product (product of heart rate and systolic blood pressure) did not show a difference between TXE and THH (P = 0.38) but was reduced compared to placebo (P = 0.0053). One subject on TXE was discontinued from the study after the first exercise test because of secondary heart block during the recovery period. It was concluded that TXE and timolol hemihydrate solution show similar systemic beta-blockade activity when both are dosed once a day.