Sensitive and selective bioanalytical assay for the determination of dobutamine in rat plasma samples |
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Authors: | H Lingeman J A Haverhals H J E M Reeuwijk U R Tjaden J van der Greef |
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Institution: | (1) Division of Analytical Chemistry, Center for Bio-Pharmaceutical Sciences, Leiden University, P. O. Box 9502, 2300 RA Leiden, The Netherlands |
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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. |
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Keywords: | Column liquid chromatography Fluorescence detection Dobutamine Plasma samples Liquid-liquid extraction |
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