Triamcinolone acetonide in ointment was determined by first-derivative ultraviolet spectrophotometry and by high performance liquid chromatography. The UV derivative spectrophotometric method eliminated the interference of the ointment excipients when the determination was made at 274 nm, Δλ = 4 nm, scan speed of 300 nm/min, and ordinate settings ±0.050. The correlation coefficient and the relative standard error of the estimate were, respectively, 0.9999 and 0.39%. The coefficient of variation and the recovery average were 0.48 and 99.77%, respectively. The chromatographic methods were standardized using a LiChrospher® 100 RP-18 (5μm) in LiChroCART® (125-4) column, UV detection at 254 nm and mobile phases consisting of: (1) acetonitrile-water (35:65 v/v) and (2) methanol-water-96% acetic acid (55:44:1 v/v), apparent pH 3.0. The peak area response versus concentration was linear at a concentration range from 10.0 to 100.0 μg/mL. The correlation coefficients and the relative standard errors of the estimate were 0.9995, 1.84% and 0.9998 and 1.14% for methods 1 and 2, respectively. The coefficients of variation and the recovery averages were 0.66%, 99.84% and 0.60%, 99.36% for methods 1 and 2, respectively.