LAUSR

The basic objective of this study is to propose a short, reliable, mass compatible ultra-performance liquid chromatography (UPLC) method to confirm the identity of impurities and to estimate the assay and purity of Tirofiban simultaneously in aqueous injection (5 mg/100 mL bag). Aqueous formulations are susceptible to oxidation, hence the possible oxidative degradation impurities of Tirofiban were studied in this experiment by using UPLC coupled with Photodiode Array/Quadrupole Dalton Analyzer (PDA/QDa) detectors. The required separations were achieved in the column: ACQUITY HSS T3 (100 x 2.1) mm, 1.7 µm, operated at 30 ºC by using 0.02 % Triethyl amine (TEA) in water, pH 2.8 with formic acid as solution-A and 0.1 % formic acid in 9:1 acetonitrile, water as solution-B. Binary gradient flow is delivered at the rate of 0.5 mL/min and the detection of impurities specifically carried out at 227nm using empower3 software. RP-UPLC/PDA with QDa detector was used for the experiment. The method was linear and accurate from the concentrations: 0.04 to 0.38 µg/mL for impurity-A and 0.04 to 75 µg/mL for Tirofiban. The major unknown degradation impurity generated during the oxidative degradation has been identified as N-oxide derivative (Impurity-B) [(M+H)+ 455.1] by using QDa detector operated in an electro spray positive ion mode by applying a voltage of 0.8kV. This method was further validated as per ICH Q2 (R2) guidelines. Hence, the proposed method is said to be a fast, sensitive and comprehensive technique which could give a clear idea about the assay and impurity profile of Tirofiban injection.