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Sensitive and selective bioanalytical assay for the determination of dobutamine in rat plasma samples
Authors:H Lingeman  J A Haverhals  H J E M Reeuwijk  U R Tjaden  J van der Greef
Institution:(1) Division of Analytical Chemistry, Center for Bio-Pharmaceutical Sciences, Leiden University, P. O. Box 9502, 2300 RA Leiden, The Netherlands
Abstract:Summary Dobutamine is one of the synthetic catecholamines acting directly onβ 1-receptors. For the analysis of dobutamine in rat plasma samples, a selective and sensitive liquid chromatographic method is described. After a simple liquid-liquid extraction, separation of the analyte was performed using a reversed-phase ion-pair system with an octyl modified silica column. The solute was detected by fluorescence detection, applying an excitation wavelength of 285nm and an emission wavelength of 313nm. The (im)possibilities of the application of the normally used assays for the isolation, concentration and quantitation of catecholamines are discussed. By the addition of a minimum amount of modifier to the mobile phase, the selectivity of the system was increased significantly. With this method the detection limit is 9ng/ml in 0.2ml plasma samples. The application of the method is shown in rat plasma samples by measuring the concentration-time curves to establish plasma level-effect relationships for this drug.
Keywords:Column liquid chromatography  Fluorescence detection  Dobutamine  Plasma samples  Liquid-liquid extraction
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