title: Determination of ciprofloxacin and levofloxacin in human sputum collected from cystic fibrosis patients using microextraction by packed sorbent-high performance liquid chromatography photodiode array detector.
abstract: This paper reports a new, easy, cheap, and fast MEPS-HPLC-PDA method for the simultaneous analysis of ciprofloxacin and levofloxacin, two fluoroquinolones (FLQs) commonly used for the treatment of pulmonary infections in cystic fibrosis (CF) patients. The FLQs were resolved on a Discovery C8 column (250mm×4.6mm; 5μm particle size) using an isocratic elution with a run time of 15min, without further purification. The method was validated over concentrations ranging from 0.05 to 2μg/mL for both analytes in human sputum, and enrofloxacin was used as internal standard. This method was successfully tested to detect FLQs in sputum collected from CF patients. The MEPS-HPLC-PDA method was validated using biological samples collected from CF patients orally or intravenously injected with FLQs. The resultant data showed that the method is selective, sensitive and robust over range of concentrations for both FLQs. The limit of quantification of the method was 0.05μg/mL for both analytes (comparable to more complex and expensive instrument configurations), weighted-matrix-matched standard curves showed a good linearity up to 2μg/mL, and parallelism tests were also successfully assessed. The intra- and inter-day precision (RSD%) values were ≤10.4% and ≤11.1%, respectively, for all range of analysis. The intra- and inter-day trueness (Bias%) values are ranged from -11.8% to 7.25% for both antibiotic drugs. At the best of our knowledge, this is the first MEPS-HPLC-PDA based method that uses MEPS procedure for simultaneous determination of ciprofloxacin and levofloxacin in human sputum. The method was tested successfully on real sputum samples by following a conventional drug administration. Furthermore, the MEPS-HPLC-PDA based method provides more advantages to detect and analyze quickly the antibiotic drugs in biological matrices than other analytical procedures reported in literature.
authors.affiliation: University of Chieti - Pescara "G. d'Annunzio"; Department of Pharmacy, via dei Vestini 31, 66100 Chieti, Italy; Interuniversity Consortium of Structural and Systems Biology, Viale Medaglie d'Oro 305, 00136 Roma, Italy. Electronic address: m.locatelli@unich.it.
authors.forename: Marcello
authors.initials: M
authors.lastname: Locatelli
authors.affiliation: University of Chieti - Pescara "G. d'Annunzio"; Department of Pharmacy, via dei Vestini 31, 66100 Chieti, Italy.
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Ciavarella
authors.affiliation: University of Catanzaro "Magna Graecia", Department of Clinical and Experimental Medicine, Viale "S. Venuta", 88100 Catanzaro, Italy; University of Catanzaro "Magna Græcia", Inter-Regional Research Center for Food Safety & Health, Viale "S. Venuta", 88100 Catanzaro, Italy.
authors.forename: Donatella
authors.initials: D
authors.lastname: Paolino
authors.affiliation: University of Chieti - Pescara "G. d'Annunzio"; Department of Pharmacy, via dei Vestini 31, 66100 Chieti, Italy; University of Catanzaro "Magna Græcia", Inter-Regional Research Center for Food Safety & Health, Viale "S. Venuta", 88100 Catanzaro, Italy; Houston Methodist Research Institute, Department of Nanomedicine, Houston, TX 77030, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Celia
authors.affiliation: Bambino Gesù" Children's Hospital, IRCCS, Piazza Sant'Onofrio 4, 00165, Rome, Italy.
authors.forename: Ersilia
authors.initials: E
authors.lastname: Fiscarelli
authors.affiliation: Bambino Gesù" Children's Hospital, IRCCS, Piazza Sant'Onofrio 4, 00165, Rome, Italy.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Ricciotti
authors.affiliation: University of Chieti - Pescara "G. d'Annunzio", Department of Medical, Oral, and Biotechnological Sciences, Via dei Vestini 31, 66100 Chieti, Italy; Center of Excellence on Ageing, "G. d'Annunzio" University Foundation, Via L. Polacchi 11, 66100 Chieti, Italy.
authors.forename: Arianna
authors.initials: A
authors.lastname: Pompilio
authors.affiliation: University of Chieti - Pescara "G. d'Annunzio", Department of Medical, Oral, and Biotechnological Sciences, Via dei Vestini 31, 66100 Chieti, Italy; Center of Excellence on Ageing, "G. d'Annunzio" University Foundation, Via L. Polacchi 11, 66100 Chieti, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Di Bonaventura
authors.affiliation: University of Chieti - Pescara "G. d'Annunzio"; Department of Pharmacy, via dei Vestini 31, 66100 Chieti, Italy; Center of Excellence on Ageing, "G. d'Annunzio" University Foundation, Via L. Polacchi 11, 66100 Chieti, Italy.
authors.forename: Rossella
authors.initials: R
authors.lastname: Grande
authors.affiliation: Selcuk University, Department of Biology, Konya, Turkey.
authors.forename: Gokhan
authors.initials: G
authors.lastname: Zengin
authors.affiliation: University of Chieti - Pescara "G. d'Annunzio"; Department of Pharmacy, via dei Vestini 31, 66100 Chieti, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Di Marzio
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002939:Ciprofloxacin / Q000032:analysis*; D003550:Cystic Fibrosis / Q000378:metabolism*; D006801:Humans; D064704:Levofloxacin / Q000032:analysis*; D057230:Limit of Detection; D052617:Solid Phase Microextraction / Q000379:methods; D013183:Sputum / Q000737:chemistry*
chemical_list: D000900:Anti-Bacterial Agents; D002939:Ciprofloxacin; D064704:Levofloxacin
keywords: Cystic fibrosis; Levofloxacin and ciprofloxacin; MEPS–HPLC–PDA; Method development; Sample preparation; Sputum