title: Robust HPLC-MS/MS method for levofloxacin and ciprofloxacin determination in human prostate tissue.
abstract: Fluoroquinolones are the drugs of choice in the prevention of bacterial infections after transrectal ultrasound guided prostate biopsy. In order to improve assessment of antibacterial efficacy in the target tissue a simple, selective, rapid and robust HPLC-ESI-MS/MS method for the determination of levofloxacin and ciprofloxacin concentrations in human prostate bioptates was developed and validated. Preparation procedure for prostate samples (10mg) was carried out using homogenization and filtration steps. Analyses were performed within 3.5min using RP C18column in the isocratic elution mode with mobile phase composed of a mixture of 0.1% formic acid aqueous solution and 0.1% formic acid methanol solution (v/v; 79:21). The method was linear between 0.3μg/g and 15μg/g for levofloxacin and ciprofloxacin with coefficient of correlation (r) ≥0.999. The limit of detection and the limit of quantification for levofloxacin were 0.06μg/g and 0.2μg/g and for ciprofloxacin were 0.04μg/g and 0.13μg/g, respectively. Average concentrations (±SD) of levofloxacin and ciprofloxacin obtained from patients tissue were 5.4±2.2μg/g and 3.9±1.5μg/g, respectively. Additionally, during validation procedure a novel, experimental design approach was applied for the robustness study. For evaluation of analytical method robustness, Plackett-Burman design was employed and for sample preparation method robustness Fractional Factorial design was used. The developed and validated method was successfully applied to examine prostate tissue samples obtained from patients enrolled into a clinical study. Up to now, there has been no other HPLC-ESI-MS/MS method reported for the simultaneous determination of levofloxacin and ciprofloxacin in human prostatic tissue.
authors.affiliation: Medical University of Gdańsk, Department of Biopharmaceutics and Pharmacodynamics, Al. Gen. J. Hallera 107, 80-416 Gdańsk, Poland.
authors.forename: O
authors.initials: O
authors.lastname: Szerkus
authors.affiliation: Medical University of Gdańsk, Department of Biopharmaceutics and Pharmacodynamics, Al. Gen. J. Hallera 107, 80-416 Gdańsk, Poland.
authors.forename: J
authors.initials: J
authors.lastname: Jacyna
authors.affiliation: Medical University of Gdańsk, Department of Urology, Mariana Smoluchowskiego 17, 80-214 Gdańsk, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Gibas
authors.affiliation: Medical University of Gdańsk, Department of Urology, Mariana Smoluchowskiego 17, 80-214 Gdańsk, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Sieczkowski
authors.affiliation: Medical University of Gdańsk, Department of Biopharmaceutics and Pharmacodynamics, Al. Gen. J. Hallera 107, 80-416 Gdańsk, Poland.
authors.forename: D
authors.initials: D
authors.lastname: Siluk
authors.affiliation: Medical University of Gdańsk, Department of Urology, Mariana Smoluchowskiego 17, 80-214 Gdańsk, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Matuszewski
authors.affiliation: Medical University of Gdańsk, Department of Biopharmaceutics and Pharmacodynamics, Al. Gen. J. Hallera 107, 80-416 Gdańsk, Poland.
authors.forename: R
authors.initials: R
authors.lastname: Kaliszan
authors.affiliation: Medical University of Gdańsk, Department of Biopharmaceutics and Pharmacodynamics, Al. Gen. J. Hallera 107, 80-416 Gdańsk, Poland. Electronic address: markusz@gumed.edu.pl.
authors.forename: M J
authors.initials: MJ
authors.lastname: Markuszewski
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002939:Ciprofloxacin / Q000032:analysis* / Q000493:pharmacokinetics*; D024841:Fluoroquinolones / Q000032:analysis; D006801:Humans; D064704:Levofloxacin / Q000032:analysis* / Q000493:pharmacokinetics*; D057230:Limit of Detection; D008297:Male; D011467:Prostate / Q000187:drug effects*; D012015:Reference Standards; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013696:Temperature
chemical_list: D000900:Anti-Bacterial Agents; D024841:Fluoroquinolones; D002939:Ciprofloxacin; D064704:Levofloxacin
keywords: Ciprofloxacin; Design of experiments; Levofloxacin; Mass spectrometry; Method validation; Prostate tissue