Simultaneous determination of levofloxacin, gatifloxacin and moxifloxacin in serum by liquid chromatography with column switching.
Liquid chromatography with a column-switching technique was developed for simultaneous direct quantification of levofloxacin, gatifloxacin and moxifloxacin in human serum. Serum samples were injected on a LiChroCART 4-4 pre-column (PC) filled with a LiChrospher 100 RP-18, 5 microm where fluoroquinolones (FQs) were purified and concentrated. The FQs were back-flushed from the PC and then separated on a Supelcosil ABZ+ Plus (150 mm x 4.6 mm i.d.) analytical column with a mobile phase containing 10 mM phosphate buffer (pH 2.5), acetonitrile (88:12, v/v) and 2mM tetrabutyl ammonium bromide. The effects of ion-pair reagents, buffer type, pH and acetonitrile concentrations in the mobile phase on the separation of the three FQs were investigated. Fluorescence detection provided sufficient sensitivity to achieve a quantification limit of 125 ng/ml for levofloxacin and moxifloxacin; 162.5 ng/ml for gatifloxacin with a 5 microl sample size. The on-line process of extraction avoids time-consuming treatment of the samples before injection and run time is shortened. The recovery, selectivity, linearity, precision and accuracy of the method are convenient for pharmacokinetic studies or routine assays.
Nguyen
Hoang Anh
HA
Laboratoire de Pharmacocinétique et de Pharmacie Clinique, Faculté de Pharmacie, Université Victor Segalen Bordeaux 2, Zone Nord, 146 Rue Léo-Saignat, 33076 Bordeaux Cedex, France.
Grellet
Jean
J
Ba
Boubakar B
BB
Quentin
Claudine
C
Saux
Marie-Claude
MC
D000890:Anti-Infective Agents / Q000097:blood*; D001372:Aza Compounds / Q000097:blood*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D024841:Fluoroquinolones / Q000097:blood*; D000077734:Gatifloxacin; D006801:Humans; D006863:Hydrogen-Ion Concentration; D064704:Levofloxacin; D000077266:Moxifloxacin; D015242:Ofloxacin / Q000097:blood*; D011786:Quality Control; D011804:Quinolines / Q000097:blood*; D012015:Reference Standards; D015203:Reproducibility of Results
D000890:Anti-Infective Agents; D001372:Aza Compounds; D024841:Fluoroquinolones; D011804:Quinolines; D064704:Levofloxacin; D015242:Ofloxacin; D000077734:Gatifloxacin; D000077266:Moxifloxacin