Two highly sensitive chromatographic methods were developed for the estimation of Tirofiban from infusion. The first method is HPLC, in which sodium dihydrogen phosphate (pH 5): Acetonitrile, 70:30% v/v was used as mobile phase and separation carried out on a C18 column. With 1 ml/min flow rate, tirofiban was eluted at 4.2min. The second method is HPTLC using a mobile phase consisting of hexane: methanol: acetic acid (5:3.5:0.5% v/v/v) on a precoated silica gel 60F254 plate. The drug was retained at a Rf of 0.46. Two analytical methods were validated for linearity, accuracy, precision, LOD, LOQ, ruggedness and robustness as per ICH guidelines. The linear concentration was 600-1400 ng/ml for HPLC and 3-18 ng/spot for HPTLC method. The LOD concentration was found to be 90 ng/ml and 1 ng/spot for HPLC and HPTLC, respectively The LOQ of Tirofiban is 400 ng/ml by HPLC and 3 ng/spot by HPTLC. The precision and accuracy values were found to be in acceptable limits. The method was successfully applied to Tirofiban infusion and the amount estimated was close to label claim. Further no interference from additives by both the methods.
