The simultaneous separation and determination of five quinolone antibiotics using isocratic reversed-phase HPLC: application to stability studies on an ofloxacin tablet formulation

A rapid and reliable HPLC method was developed for the simultaneously separation and quantitation of five quinolones antibiotics; nalidixic acid, norfloxacin, ofloxacin, ciprofloxacin and lomefloxacin. All five tablet formulations of individual quinolone antibiotics were routinely assayed without interference. The calibration curves were linear (r2 ≥ 0.999) over the concentration range of 1.20–4.8 mg/100 ml. Selectivity, precision, sensitivity and accuracy were established and the method is stability indicating with respect to ofloxacin. The limit of detection and quantitation for ofloxacin was 18 and 36 μg/100 ml, respectively. The separation was performed on a Phenomenex ODS C18 column using an isocratic, ion-pairing mobile phase consisting of 35% (v/v) aqueous acetonitrile together with tetrabutylammonium acetate, sodium dodecyl sulphate and citric acid (pH* 3.4). All analyses were conducted at ambient temperature and was monitored using a Diode Array UV/VIS detector set at wavelengths 235, 254, 275 and 300 nm.