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A new, rapid, sensitive and selective HPLC method with fluorescence detection is described for the simultaneous determination of 12 sulfonamides, in the presence of putrescine as internal standard, after pre-column derivatization with fluorescamine. The drugs were separated on a Chromolith Performance RP-18 column (100x4.6 mm), using a gradient elution with a binary mobile phase of methanol/0.05 M acetate buffer (pH 3.4). Linearity of derivatization was obtained for concentrations from 3.0 to 300 microg/L in standard solutions. The whole procedure was evaluated and fully validated, according to the European Union Decision 2002/657/EC, for the determination of sulfonamides in turkey muscle and hen eggs following SPE. The LODs varied from 2 to 17 microg/kg in turkey and 2 to 15 microg/kg in egg samples. The average recoveries ranged between 96.9-108.6% in turkey muscle and 96.0-108.4% in egg samples, respectively.