A simple, rapid, accurate and sensitive method was developed for quantitative analysis of Trospium Chloride in tablet dosage form using Reverse Phase high performance liquid chromatography (RP-HPLC) with DAD detection. The chromatography separation was achieved Xterra RP18, (250 x 4.6mm, 5µParticle Size)), at column temperature 40°C±2, in the isocratic mobile phase mode using Acetonitrile (ACN): Buffer (65:35; v/v) at a flow rate of 1.0mL/min. The determination was performed using Buffer HPLC with DAD detector set at 215nm. Samples were prepared with diluent [Buffer: ACN (200:100; v/v)], and the volume injected was 50µL. The retention time for Trospium Chloride was 3.275 minute. The analytical curve was linear (r2 0.9993) over a wide concentration range (50.0-150.0 µg/mL). The presence of components of the tablet did not interfere in the results of the analysis. The method showed adequate precision with a relative standard deviation (RSD) less than 2%.