A new, sensitive, and selective high-performance liquid chromatography (HPLC) method with fluorimetric detection was developed for the determination of moxifloxacin (MOX) in human breast milk. MOX was precolumn derivatized with fluorescamine; the fluorescent derivative was separated on an RP C18 column using a mobile phase composed of acetonitrile–10 mM orthophosphoric acid by isocratic elution with flow rate of 0.5 mL min−1. The method was based on the measurement of the derivative using fluorescence detection at 481 nm with excitation at 351 nm. The calibration curve was linear over the range of 1–40 μg mL−1. Limit of detection (LOD) and limit of quantitation (LOQ) were found to be 0.3 and 1 μg mL−1, respectively. Intra-day and interday repeatabilities were less than 3.15%.
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