title: Quantitative determination of azithromycin in plasma, blood and isolated neutrophils by liquid chromatography using pre-column derivatization with 9-fluorenylmethyloxycarbonyl-chloride and fluorescence detection.
abstract: In this study, a high-performance liquid chromatographic method with pre-column derivatization and fluorescence detection was optimised and validated for the quantification of azithromycin (AZM) in plasma. Clarithromycin (CLM) was used as an internal standard. Pre-column derivatization was done with 9-fluorenylmethyloxycarbonyl-chloride. Recovery from blood and polymorphonuclear neutrophils (PMNNs) isolated by a gravity separation procedure was also assessed. Analytical separation was carried out using a C18 column as stationary phase and acetonitril-phosphatebuffer as mobile phase. Peak quantification was carried out by excitation at 26 7 nm and detection at 317 nm. A lower limit of quantitation of 0.042+/-0.017 mg/l in plasma, 0.119+/-0.065 mg/l in blood and 0.072+/-0.036 in water was achieved. Linearity was assessed from 0 to 1.5mg/l in plasma and blood and from 0-9 mg/l in water. The analytical method proved to be applicable in a pharmacokinetic study of AZM in a Cystic Fibrosis patient.
authors.affiliation: The Hague Central Hospital Pharmacy, Escamplaan 900, The Hague 2504 AC, The Netherlands. e.wilms@ahz.nl
authors.forename: Erik
authors.initials: E
authors.lastname: Wilms
authors.forename: Henk
authors.initials: H
authors.lastname: Trumpie
authors.forename: Wim
authors.initials: W
authors.lastname: Veenendaal
authors.forename: Daan
authors.initials: D
authors.lastname: Touw
mesh_terms: D000900:Anti-Bacterial Agents / Q000097:blood* / Q000493:pharmacokinetics; D017963:Azithromycin / Q000097:blood* / Q000493:pharmacokinetics; D005449:Fluorenes / Q000737:chemistry*; D006801:Humans; D009504:Neutrophils / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: C496789:9-fluorenylmethoxycarbonyl; D000900:Anti-Bacterial Agents; D005449:Fluorenes; D017963:Azithromycin