Development and validation of reverse phase high performance liquid chromatographic method for determination of Tirofiban in serum

А 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

Development of alternative HPLC method for the determination of Tirofiban in rat serum

Tirofiban hydrochloride is a reversible antagonist of fibrinogen binding to the GPIIb/IIIa receptor, used for the treatment of acute coronary syndrome. A novel RP-HPLC method has been developed and validated for the determination of Tirofiban in serum of Wistar rats with and without deep acute venous thrombosis. The chromatographic separation was carried out using a reverse-phase HPLC column Puro-spher® RP-18e (150 mm 4.6 mm i.d.; 5 μm) coupled with a guard column LiChrosorb® (4 mm 4 mm i.d.; 7 μm) and mobile phase consisting of the mixture of 1-octane sulfonic acid in water (pH 3.0, adjusted with orthophosphoric acid) and acetonitrile, with a ratio of 60 : 40 (v/v) and a flow rate of 1.0 ml/min, at a wavelength of 277 nm. The serum concentrations of Tirofiban in the group of rats with DVT were lower than those in the control group, and it could be explained with the binding of Tirofiban with the GPIIb/IIIa receptors

Imaging of deep venous thrombosis using radioactive-labeled tirofiban

The development of radiolabeled small peptide or peptidomimetic ligands can bind platelets and their specific expressed receptor have been suggested as a new approach to detect the clot location and, more essentially, to determine the age and morphology of the evolving thrombus. This new approach is focused on the use of a series of radiolabeled platelet GPIIb/IIIa receptor antagonists. Tirofi ban N-(butylsulfonyl)- 4-O-(4-(4-piperidyl)-L-tyrosine is a non-peptide tyrosine derivate. The aim of the study was to introduce radioactive-labeled tirofi ban as a specifi c imaging agent for acute DVT. The labeling was performed with technetium-99 in the presence of a stannous reducing agent. The labeled preparation showed fast blood clearance in a normal rat model (without induced thrombosis). More than 80 % of the injected dose was eliminated from the circulation in the fi rst hour after injection. Biodistribution and visualization of the labeled molecule was carried out using an experimental model of thrombosis in a male Wistar rat. Planar images were obtained 30 and 60 min after application of 2 × 106 imp/min 99m-technetium-tirofi ban in the rat’s tail vein. Sensitivity and specifi city were determined using the ratio of ‘left leg positive for DVT’ to ‘right leg negative for DVT’. The obtained ratio was 1.54 after 30 min and 5.04 after 60 min. These values were considered positive in the detection of acute DVT. The high DVT uptake shows that radiolabeled tirofi ban in the introduced rat model can be a promising agent for imaging the deep venous thrombosis

Imaging of deep venous thrombosis using radioactive labeled Tirofiban: animal model evaluation

Imaging of acute thrombus, especially the very prevalent condition of acute deep vein thrombosis is usually relied on conventional imaging techniques utilizing either ultrasonography or contrast venography. The former procedure is limited by accuracy and the latter by technical considerations. Recent advances in the understanding of the pathogenesis of acute clot at the molecular level have suggested new opportunities for detection of the acute thrombotic process based on the biomolecular behavior of components of the clotting process including the formed element of the blood, the platelet. Thus, development of radiolabelled small peptide or peptidomimetic ligands that can bind platelets and their specific expressed receptor have been suggested as a new approach to detect clot location and, more essentially, determine the age and morphology of the evolving thrombus. This new approach has focused on the use of a series of radiolabelled platelet GPIIb/IIIa receptor antagonists. Tirofiban N-(butylsulfonyl)- 4-O-(4-(4-piperidyl)-L-tyrosine is a non-peptide tyrosine derivate. The aim of the study was to introduce radioactive labelled tirofiban as a specific imaging agent for acute DVT. The labeling was performed with Technetium-99 in the presence of a stannous reducing agent. The labelled preparation showed a fast blood clearance in the normal rat model (without induced thrombosis). More than 80% of the injected dose was eliminated from the circulation in the first hour after injection. Biodistribution and visualization of the labelled molecule was carried out using an experimental model of thrombosis in the male Wistar rat. Planar images were obtained 30 min and 60 min after application of 2 × 106 imp/min 99mTechnetium-tirofiban, in the rat’s tail vein. Sensitivity and specificity were determined using the ratio ‘left leg positive for DVT’ and ‘right leg negative for DVT’. The obtained ratio was 1.54 after 30 min and 5.04 after 60 min. These values were considered as positive in the detection of acute DVT. The high DVT uptake show that radiolabelled tirofiban in the introduced rat model can be the promising agent for imaging of deep venous thrombosis

Determination of tirofiban in serum using liquid chromatography with UV detection

The aim of this study is to develop a specific, sensitive, rapid, and simple HPLC method with UV detection to study pharmacokinetics of tirofiban in serum of rats alone or in the presence of heparin as simultaneous therap

Serum determination of 99m Technetium radiolabeled Tirofiban using high performance liquid chromatography in the animal rat model of introduced acute deep venous thrombosis

The development of radiolabeled small peptide or peptidomimetic ligands can bind platelets and their specific expressed receptor have been suggested as a new approach to detect the clot location and, more essentially, to determine the age and morphology of the evolving thrombus. This new approach is focused on the use of a series of radiolabeled platelet GPIIb/IIIa receptor antagonists. Tirofiban N-(butylsulfonyl)- 4-O-(4-(4-piperidyl)-L-tyrosine is a non-peptide tyrosine derivate. The aim of the study was to introduce radioactive-labeled tirofiban as a specific imaging agent for acute DVT and to determine the serum concentrations in normotensive male Wister rats with and without deep acute venous thrombosis in order to confirm the animal model of acute venous thrombosis