Abstract
A new, simple, specific, accurate and precise reversed phase-high performance liquid chromatography (RP-HPLC) method was developed for the determination of levofloxacin in rat plasma and saliva was developed. An HPLC system based on a Phenomenex Luna C18Column (250 × 4.6 mm) and a UV detector (λ = 296 nm) were used. A mixture of Acetonitrile: water (80:20 v/v) adjusted to pH 3.5 by orthophosphoric acid at a flow rate of 1.4 ml/min was used as mobile phase. The proteins were precipitated with methanol. The average recovery was 94.79 and 92.66%, respectively in plasma and saliva. The detection limit for levofloxacin in plasma and saliva was 1 µg/ml. The calibration curve was linear over the concentration range 1 to 16 mg/ml for plasma and saliva. The inter-day and intra-day assay coefficients of variation were found to be less than 5%. The present validated method was successfully used for pharmacokinetic studies of levofloxacin in plasma and saliva. Key words: High-performance liquid chromatography (HPLC), levofloxacin, plasma, saliva, pharmacokinetics.
Highlights
Levofloxacin (LOFLX), (–)-(S)-9-fluoro-2,3-dihydro-3methyl-10-(4-methyl-1-piperazinyl)-7-oxo-7H-pyrido[1,2,3 -de]-1,4-benzoxazine-6-carboxylic acid, is an advancedgeneration fluoroquinolone antibiotic
The respective regression equations for plasma and saliva were y = 238950.2x + 5607.514 (r = 0.999) and y = 276625.2x + 25097.29 (r = 0.999), where y is the peak-area of the drug to the internal standard, x is the concentration in plasma or saliva and r is the coefficient of correlation, and that statistically confirms the linearity of this method
We assessed the precision of the method by repeated analysis of plasma specimens containing known concentrations of LOFLX
Summary
Levofloxacin (LOFLX), (–)-(S)-9-fluoro-2,3-dihydro-3methyl-10-(4-methyl-1-piperazinyl)-7-oxo-7H-pyrido[1,2,3 -de]-1,4-benzoxazine-6-carboxylic acid, is an advancedgeneration fluoroquinolone antibiotic. It has broadspectrum in vitro activity, including activity against many clinically encountered Gram positive and Gram negative organisms (Fu et al, 1992), and is administered to treat various infectious diseases, like community acquired and nosocomial pneumonia, skin and skin structure infection, urinary tract infections or sepsis (North et al, 1998). Several HPLC methods coupled with different detection techniques have been reported for quantification of levofloxacin in blood plasma (Siewert, 2006; Djabarouti et al, 2004). It has been reported that the lipophilicity of the fluoroquinolones primarily determines the extent of salivary excretion (Langlois et al, 2005) and LOFLX has intermediate lipophilicity (Andersson and MacGowan, 2003).
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