To ensure that residues of Tirofiban will not carry over and cross contaminate the subsequent product, carefully designed cleaning validation techniques should be investigated. The objective of the present study was to validate simple analytical method for verification of residual Tirofiban in equipments used in the production area and to confirm efficiency of cleaning procedure. An assay method for the determination of Tirofiban residues on stainless steel surface is proposed. The cleaning validation procedure for the equipment was done by the use of cotton swabs moistened with the extraction solution 800 ml of water and 1.36 g of Ammonium acetate and then adjusts at pH (6.0 ± 0.05). In this method symmetry C18 5 μm is utilized as the stationary phase in the HPLC column, (250×0.4 mm) at 40°C using acetonitrile- buffer 6 (29-71%) mixture as mobile phase with injected volume 100 μl with flow rate 1.5 ml/min over the concentration range 1.0-2.9 μg/mL were determined at 227 nm using UV spectrophotometer detector. The recover were 107.96, 100 and 88.78% with RSD below 1.5% for the three assayed concentration levels and with average recovery was 98.89%.