Quantitative determination of besifloxacin, a novel fluoroquinolone antimicrobial agent, in human tears by liquid chromatography–tandem mass spectrometry

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 867; no. 1; pp. 105 - 110
• Main Authors: Arnold, Dana R., Granvil, Camille P., Ward, Keith W., Proksch, Joel W.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.05.2008; Elsevier Science
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Anti-Bacterial Agents - analysis; Anti-Bacterial Agents - pharmacokinetics; Antimicrobial agent; Azepines - analysis; Azepines - pharmacokinetics; Besifloxacin; Biological and medical sciences; Calibration; Chromatography, High Pressure Liquid - methods; Fluoroquinolone; Fluoroquinolones - analysis; Fluoroquinolones - pharmacokinetics; Fundamental and applied biological sciences. Psychology; General pharmacology; Human tears; Humans; LC/MS/MS; Medical sciences; Pharmacokinetics; Pharmacology. Drug treatments; Reference Standards; Reproducibility of Results; Sensitivity and Specificity; Tandem Mass Spectrometry - methods; Tears - chemistry; Fluoroquinolone; Human tears; Pharmacokinetics; LC/MS/MS; Besifloxacin; Antimicrobial agent; Human; Fluoroquinolone derivatives; Tear; Liquid chromatography; Antibacterial agent; Besifloxacin,Fluoroquinolone,Antimicrobial agent,Human tears,LC/MS/MS,Pharmacokinetics; Quantitative analysis
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2008.03.025

A rapid and sensitive method was developed using high-performance liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) for the quantification of besifloxacin in human tears using sparfloxacin as the internal standard (IS). Besifloxacin was extracted from human tear samples using an ammonium formate buffer at pH 3.25. The method was validated over a concentration range of 2–2000 ng/mL, with a total run time of less than 4 min. The overall intra- and inter-day precision for this method was less than 6%. The method was used to measure besifloxacin concentrations in tear samples collected after topical ocular administration to humans; besifloxacin concentrations were 610 ± 540 μg/g (15 min) and 1.60 ± 2.28 μg/g (24 h).