Journal of Chromatography B: Biomedical Sciences and Applications
Determination of moxifloxacin in growth media by high-performance liquid chromatography
Introduction
Moxifloxacin (BAY 12-8039) is a new fluoroquinolone with a broad spectrum of activity encompassing gram-negative and gram-positive bacteria [1]. The mechanism of acquired resistance to the quinolones is consistent among currently available drugs in this class and is expected to be similar for new and developmental agents as well. In the laboratory, selection of bacterial resistance depends on the particular quinolone, its concentration, and the species of bacterium involved [2]. Several in vitro pharmacokinetic–pharmacodynamic (PK–PD) models were developed to evaluate the efficacy of antimicrobial agents and possible emergence of resistance. These models allows for comparisons of antimicrobial efficacy of newly developed antibiotics with the efficacy of older drugs. Mutation rates in the presence of clinically relevant concentration time curves may be compared for various drugs and regimens [3]. Such experiments are time-consuming because of the need to examine a large number of samples. Therefore, rapid analytical methods are required for labor-saving.
High-performance liquid chromatography (HPLC) methods involving deproteinization of human body fluids samples followed by centrifugation [4], [5] or solid-phase extraction [6] and a capillary electrophoresis technique [7] were proposed for moxifloxacin determination. A new assay using column switching was developed to allow on-line HPLC by direct injection of Mueller–Hinton broth (M.–H.B.) sample, shortening greatly the sample processing. The on-line method consists of a first step of trapping the analyte in the pre-column (PC) and elution of biological matrix to the waste. In a second step, the analyte is transferred to the analytical column (AC) and separation occurs.
Section snippets
Chemicals
Moxifloxacin hydrochloride was a gift from Bayer (Puteaux, France). Compounds obtained from pharmaceutical companies and used for analytical interference studies were: amikacine, cefepim (Bristol-Myers Squibb, Paris, France), tobramycine (Lilly, Saint Cloud, France), gentamicine (Dakota, Paris, France), cefotaxime, cefpirome (Roussel, Paris, France), ceftazidime (Glaxo Wellcome, Marly-le-Roi, France) and ceftriaxone (Roche, Neuilly-sur-Seine, France). Analytical-grade (Normapur) dipotassium
HPLC system
In order to assay many experimental samples of moxifloxacin, we adopted direct injection with a column switching technique which was practical without the disadvantages of classical labor-intensive and time consuming preparation process [8], [9]. The pre-column and the 5 μm filter were replaced after injection of about 100 samples. All the development and validation tests, that is to say at least 500 samples were analyzed with the same analytical column without appreciable decrease of its
Application
The method described was successfully applied to the analysis of M.–H.B. samples from an in vitro PK–PD model derived from the two-compartment kinetic model with artificial capillary units proposed by Blaser et al. [13]. The model was designed to expose bacteria to changing antibiotic concentrations, without dilution of the bacterial inoculum together with the antibiotic. The central compartment (CCp) consisted of a thermostatizable flask with magnetic stirrer containing culture broth, tubing
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Moxifloxacin Hydrochloride
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2008, Spectrochimica Acta - Part A: Molecular and Biomolecular SpectroscopyHigh-performance liquid chromatography assay for moxifloxacin: Pharmacokinetics in human plasma
2007, Journal of Pharmaceutical and Biomedical AnalysisOptimization and validation of the direct HPLC method for the determination of moxifloxacin in plasma
2006, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences