A novel sensitive analytical method based on the use of a semi-automatic microextraction by packed sorbents (MEPS) techniques combined with ultra high-performance liquid chromatography (UHPLC) with PDA detection has been developed and validate for the analysis of ulifloxacin, the active metabolite of prulifloxacin using danofloxacin as internal standard in human plasma and urine. Different experimental parameters were optimized and validated according to international guidelines. Complete separation of the analytes was achieved with a Waters BEH C18 (50 × 2.1 mm I.D., 1.7 μm particle size) analytical column, a mixture of 10 mM ammonium acetate (pH 3.0) (A) with and acetonitrile (B) both containing 1% triethylamine were used as mobile phase, at a flow rate of 0.6 mL/min in gradient elution, and detection wavelength of 272 nm. This method is linear in concentration range of 0.02-10.0 μg/mL for plasma and urine, respectively. The limit of quantitation was 20 ng/mL for the two fluids. The recoveries of the method were 95% for ulifloxacin in human plasma and urine and 95.5% for the internal standard. Intra- and inter- assay precision and accuracy for ulifloxacin were lower than 10% at all tested concentrations. The proposed method was successfully applied to measure plasma and urine concentrations of ulifloxacin in patients suffering from Peripheral Arterial Disease and for pharmacokinetics study.

Development and validation of a MEPS-UHPLC-PDA method for determination of ulifloxacin in human plasma and urine of patients with peripheral arterial disease

Palumbo, Paola;Carlucci, Giuseppe
2016-01-01

Abstract

A novel sensitive analytical method based on the use of a semi-automatic microextraction by packed sorbents (MEPS) techniques combined with ultra high-performance liquid chromatography (UHPLC) with PDA detection has been developed and validate for the analysis of ulifloxacin, the active metabolite of prulifloxacin using danofloxacin as internal standard in human plasma and urine. Different experimental parameters were optimized and validated according to international guidelines. Complete separation of the analytes was achieved with a Waters BEH C18 (50 × 2.1 mm I.D., 1.7 μm particle size) analytical column, a mixture of 10 mM ammonium acetate (pH 3.0) (A) with and acetonitrile (B) both containing 1% triethylamine were used as mobile phase, at a flow rate of 0.6 mL/min in gradient elution, and detection wavelength of 272 nm. This method is linear in concentration range of 0.02-10.0 μg/mL for plasma and urine, respectively. The limit of quantitation was 20 ng/mL for the two fluids. The recoveries of the method were 95% for ulifloxacin in human plasma and urine and 95.5% for the internal standard. Intra- and inter- assay precision and accuracy for ulifloxacin were lower than 10% at all tested concentrations. The proposed method was successfully applied to measure plasma and urine concentrations of ulifloxacin in patients suffering from Peripheral Arterial Disease and for pharmacokinetics study.
File in questo prodotto:
File Dimensione Formato  
1-s2.0-S0731708516303181-main.pdf

solo utenti autorizzati

Tipologia: Documento in Pre-print
Licenza: Creative commons
Dimensione 862.21 kB
Formato Adobe PDF
862.21 kB Adobe PDF   Visualizza/Apri   Richiedi una copia

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11697/132699
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 14
  • ???jsp.display-item.citation.isi??? 13
social impact