Quantitative determination of gatifloxacin in tablets, solid lipid nanoparticles (SLNs) and eye-drops using a very simple and rapid chromatographic technique was validated and developed. Formulations were analyzed using a reverse phase SUPELCO® 516 C-18-DB, 50306-U, HPLC column (250mm×4.6mm, 5μm) and a mobile phase consisting of disodium hydrogen phosphate buffer:acetonitrile (75:25, v/v) and with orthophosphoric acid pH was adjusted to 3.3 The flow rate was 1.0mL/min and analyte concentrations were measured using a UV-detector at 293nm. The analyses were performed at room temperature (25±2°C). Gatifloxacin was separated in all the formulations within 2.767min. There were linear calibration curves over a concentration range of 4.0–40μg.mL −1 and correlation coefficients of 0.9998 with an average recovery above 99.91%. Detection of analyte from different dosage forms at the same R t indicates the specificity and stability of the developed method.