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Abstract
The aim of this study was to develop a rapid and sensitive method for the simultaneous
determination of unbound levofloxacin in rat blood and bile using high-performance
liquid chromatography coupled with microdialysis for further pharmacokinetic study.
Microdialysis probes were simultaneously inserted into the jugular vein toward the
right atrium and the bile duct of male Sprague-Dawley rats for biological fluid sampling
after administration of levofloxacin 3 mg/kg through the femoral vein. Levofloxacin
and dialysates were separated using a Merck LiChrospher reversed-phase C18 column
maintained at ambient temperature. The mobile phase was comprised of acetonitrile-1
mM 1-octanesulfonic acid (40:60, v/v, pH 3.0 adjusted with orthophosphoric acid).
The fluorescence response for levofloxacin was observed at excitation and emission
wavelengths of 292 and 494 nm, respectively. The detection limit of levofloxacin was
50 ng/ml. Intra-day and inter-day precision and accuracy of levofloxacin measurements
fell well within the predefined limits of acceptability. The disposition of levofloxacin
in the blood and bile fluid suggests that there was rapid exchange and equilibration
between the blood and hepatobiliary systems, and the plasma level of levofloxacin
was greater than that of the bile. Thus, levofloxacin undergoes hepatobiliary excretion
but might not be related to the P-glycoprotein transport system.