Development and Validation of Reverse Phase High Performance Liquid Chromatographic Method for Determination of Tirofiban in Serum

A specific, sensitive and rapid RP-HPLC method has been developed for the determination of Tirofiban in serum. The chromatographic separation was realized using reverse phase LiChrospher ® 100 RP-18 column (4.0 mm × 250 mm, 5 μm) and mobile phase consisting the mixture of 0.1 M KH2PO4 (pH 5.2, adjusted with 1.0 N sodium hydroxide solution) and acetonitrile, with the ratio of 70:30% (v/v) and flow rate of 1.0 ml/min. The detection was carried out at 274 nm. The response was linear over the range of 0.03 – 0.18 mgmL -1 in mobile phase and serum samples. The limit of detection (LOD) for Tirofiban was 1.84, 13.8 and 14.6 μg mL-1 in methanol, spiked rat serum and spiked human serum, respectively. The described method can be quickly and routinely applied, without any interference from endogenous substances, for therapeutic monitoring of levels of Tirofiban in the serum samples.