Determination of ceftazidime in dolphin serum by liquid chromatography with ultraviolet-visible detection and confirmation by thermospray liquid chromatography-mass spectrometry. 1992

K L Tyczkowska, and S S Seay, and M K Stoskopf, and D P Aucoin
Department of Anatomy, Physiological Sciences and Radiology, College of Veterinary Medicine, North Carolina State University, Raleigh 27606.

A simple and sensitive liquid chromatographic method has been developed for the determination of therapeutic levels of ceftazidime in dolphin serum. The method involved an ultrafiltration of diluted serum with an equal amount of acetonitrile-ethanol-water (40:40:20, v/v/v) through a 10,000 daltons molecular mass cut-off filter. Separation of ceftazidime from the other serum components was performed by ion-paired (dodecanesulfonate) liquid chromatography using a reversed-phase column eluted with acetonitrile-water solution. The ultraviolet absorbance of the column effluent was monitored in the 200-340 nm range of a photodiode-array detector or at 258.8 nm on a variable-wavelength ultraviolet-visible detector. Recoveries of ceftazidime from dolphin serum spiked with 20 and 2 micrograms/ml were 92.9 and 91.1% with coefficients of variation of 5.5 and 5.7%, respectively. A correlation coefficient of 0.9994 occurred with ceftazidime in aqueous solutions (n = 6, in duplicates). The limit of detection for this antibiotic was estimated to be approximately 50 ppb (ng/ml). The unbound ceftazidime concentrations in dosed dolphin serum were determined to calculate the protein bindings of this antibiotic which yielded 32 +/- 2%. The ceftazidime peak identity in dosed dolphin serum was confirmed by thermospray liquid chromatography-mass spectrometry. The thermospray mass spectrum of ceftazidime exhibited only the fragment ions, involving the opening of the beta-lactam ring, at m/z 237, 255 and 315 when positive-ion detection mode was employed and the fragment ions at m/z 235, 253 and 313 when negative-ion detection mode was used.

UI MeSH Term Description Entries
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D001798 Blood Proteins Proteins that are present in blood serum, including SERUM ALBUMIN; BLOOD COAGULATION FACTORS; and many other types of proteins. Blood Protein,Plasma Protein,Plasma Proteins,Serum Protein,Serum Proteins,Protein, Blood,Protein, Plasma,Protein, Serum,Proteins, Blood,Proteins, Plasma,Proteins, Serum
D002442 Ceftazidime Semisynthetic, broad-spectrum antibacterial derived from CEPHALORIDINE and used especially for Pseudomonas and other gram-negative infections in debilitated patients. Ceftazidime Anhydrous,Ceftazidime Pentahydrate,Fortaz,Fortum,GR-20263,LY-139381,Pyridinium, 1-((7-(((2-amino-4-thiazolyl)((1-carboxy-1-methylethoxy)imino)acetyl)amino)-2-carboxy-8-oxo-5-thia-1-azabicyclo(4.2.0)oct-2-en-3-yl)methyl)-, inner salt, pentahydrate, (6R-(6alpha,7beta(Z)))-,Tazidime,GR 20263,GR20263,LY 139381,LY139381
D002853 Chromatography, Liquid Chromatographic techniques in which the mobile phase is a liquid. Liquid Chromatography
D004289 Dolphins Mammals of the families Delphinidae (ocean dolphins), Iniidae, Lipotidae, Pontoporiidae, and Platanistidae (all river dolphins). Among the most well-known species are the BOTTLE-NOSED DOLPHIN and the KILLER WHALE (a dolphin). The common name dolphin is applied to small cetaceans having a beaklike snout and a slender, streamlined body, whereas PORPOISES are small cetaceans with a blunt snout and rather stocky body. (From Walker's Mammals of the World, 5th ed, pp978-9) Amazon Dolphins,Baiji,Cephalorhynchus,Chinese River Dolphin,Delphinidae,False Killer Whale,Franciscana,Fraser's Dolphin,Ganges Dolphin,Gray Grampus,Humpback Dolphins,Irrawaddy River Dolphin,La Plata Dolphin,Lagenorhynchus,Lissodelphis,Many-Toothed Blackfish,Marine Dolphins,Melon-Headed Whale,Ocean Dolphins,Piebald Dolphins,Platanista,Pontoporia blainvillei,Pygmy Killer Whale,Right Whale Dolphins,Risso's Dolphin,Rough-Toothed Dolphin,Sousa,Susus,Tucuxi Dolphin,Whale, False Killer,Whale, Melon-Headed,Whale, Pygmy Killer,White-Beaked Dolphins,Amazon River Dolphins,Atlantic White-Sided Dolphins,Feresa attenuata,Ganges River Dolphin,Grampus griseus,Inia geoffrensis,Lagenodelphis hosei,Lagenorhynchus acutus,Lagenorhynchus albirostris,Langenorhynchus obliquidens,Lipotes vexillifer,Orcaella brevirostris,Pacific White-Sided Dolphins,Peponocephala electra,Planista gangetica,Pseudorca crassidens,Sotalia fluviatilis,Steno bredanensis,Yangtze River Dolphin,Atlantic White Sided Dolphins,Atlantic White-Sided Dolphin,Baijus,Dolphin,Dolphin, Fraser's,False Killer Whales,Franciscanas,Fraser Dolphin,Frasers Dolphin,Humpback Dolphin,Lagenodelphis hoseus,Lagenorhynchus albirostri,Langenorhynchus obliquiden,Lissodelphi,Many Toothed Blackfish,Marine Dolphin,Melon Headed Whale,Melon-Headed Whales,Ocean Dolphin,Orcaella brevirostri,Pacific White Sided Dolphins,Pacific White-Sided Dolphin,Piebald Dolphin,Planista gangeticas,Platanistas,Pontoporia blainvilleus,Pseudorca crassiden,Pygmy Killer Whales,Right Whale Dolphin,Risso Dolphin,Rissos Dolphin,Rough Toothed Dolphin,Rough-Toothed Dolphins,Sotalia fluviatili,Tucuxi Dolphins,White Beaked Dolphins,White-Beaked Dolphin
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013056 Spectrophotometry, Ultraviolet Determination of the spectra of ultraviolet absorption by specific molecules in gases or liquids, for example Cl2, SO2, NO2, CS2, ozone, mercury vapor, and various unsaturated compounds. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Ultraviolet Spectrophotometry
D013058 Mass Spectrometry An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers. Mass Spectroscopy,Spectrometry, Mass,Spectroscopy, Mass,Spectrum Analysis, Mass,Analysis, Mass Spectrum,Mass Spectrum Analysis,Analyses, Mass Spectrum,Mass Spectrum Analyses,Spectrum Analyses, Mass
D014462 Ultrafiltration The separation of particles from a suspension by passage through a filter with very fine pores. In ultrafiltration the separation is accomplished by convective transport; in DIALYSIS separation relies instead upon differential diffusion. Ultrafiltration occurs naturally and is a laboratory procedure. Artificial ultrafiltration of the blood is referred to as HEMOFILTRATION or HEMODIAFILTRATION (if combined with HEMODIALYSIS).

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