Determination of moxifloxacin in growth media by high-performance liquid chromatography

doi:10.1016/S0378-4347(00)00591-0

Journal of Chromatography B: Biomedical Sciences and Applications

Volume 754, Issue 1, 15 April 2001, Pages 107-112

https://doi.org/10.1016/S0378-4347(00)00591-0 Get rights and content

Abstract

A direct injection high-performance liquid chromatographic method with column switching has been developed to determine moxifloxacin in Mueller–Hinton broth. A LiChrocart 4-4 pre-column filled with a LiChrospher 100 RP 18, 5 μm and a 150×4.6 mm I.D. column packed with a Supelcozil ABZ+ Plus were used and led to a retention time of 5.70 min. Fluorescence detection allowed one to reach a quantification limit of 0.05 μg/ml with a 100-μl sample size. The standard curves were linear from 0.05 to 3.2 μg/ml. Intra- and inter-day imprecisions within the linearity range were ≤4.76 and ≤5.75%, respectively. The mean relative errors for the same day and the day-to-day inaccuracies ranged from −2.93 to +4.50% and from −1.10 to +6.00%, respectively. The method was demonstrated to be useful for pharmacokinetic–pharmacodynamic studies of moxifloxacin in an in vitro model.

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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The present report describes a rapid and sensitive ultra high-pressure liquid chromatography (UHPLC) method with UV detection to quantify moxifloxacin in rabbit aqueous humor. After deproteinisation with acetonitrile, gradient separation of moxifloxacin was achieved on a Waters Acquity BEH C18 (50 mm × 2.1 mm, 1.7 μm) column at 50 °C. The mobile phase consisted of 0.1% trifluoroacetic acid in water and acetonitrile at a flow rate of 0.4 ml/min. Detection of moxifloxacin was done at 296 nm. Method was found to be selective, linear (r = 0.9997), accurate (recovery, 97.30–99.99%) and precise (RSD, ≤1.72%) in the selected concentration range of 10–1000 ng/ml. Detection and quantitation limit of moxifloxacin in aqueous humor were 0.75 and 2.5 ng/ml, respectively. The aqueous humor levels of moxifloxacin after single topical instillation in three formulations, i.e. moxifloxacin solution (Moxi-SOL), anionic nanoparticles (Moxi-ANP) and cationic nanoparticles (Moxi-CNP) were investigated. A fourfold increase in the relative bioavailability was observed with the Moxi-CNP (AUC_0→t, 3.14 μg h/ml) compared with Moxi-SOL (AUC_0→t, 0.79 μg h/ml) and Moxi-ANP (AUC_0→t, 0.72 μg h/ml) formulation. The results indicate that the cationic nanoparticle increases ocular bioavailability of moxifloxacin and prolong its residence time in the eye.
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Simple, rapid and highly sensitive spectrofluorimetric method is presented for the determination of four fluoroquinolone (FQ) drugs, ciprofloxacin, enoxacin, norfloxacin and moxifloxacin in pharmaceutical preparations. Proposed method is based on the derivatization of FQ with 4-chloro-7-nitrobenzofurazan (NBD-Cl) in borate buffer of pH 9.0 to yield a yellow product.
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Intra-day and inter-day relative standard deviation and relative mean error values at three different concentrations were determined. The low relative standard deviation values indicate good precision and high recovery values indicate accuracy of the proposed methods.
The method is highly sensitive and specific. The results obtained are in good agreement with those obtained by the official and reference method.
The results presented in this report show that the applied spectrofluorimetric method is acceptable for the determination of the four FQ in the pharmaceutical preparations. Common excipients used as additives in pharmaceutical preparations do not interfere with the proposed method.
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Determination of moxifloxacin in growth media by high-performance liquid chromatography

Abstract

Introduction

Section snippets

Chemicals

HPLC system

Application

J. Chromatogr. B

J. Chromatogr. B

J. Chromatogr. B

J. Chromatogr. B

J. Chromatogr. B

J. Chromatogr. B

Drugs

Clin. Ther.