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A new and sensitive pre-column derivatization method was developed for the analysis of melamine leached from tableware by high performance liquid chromatography (HPLC) with fluorescence detection. The HPLC sensitivity was greatly enhanced by introducing 10-methyl-acridone-2-sulfonyl chloride (MASC) with excellent fluorescence property into the melamine molecule. Meanwhile, derivatization also greatly increased the hydrophobicity of melamine. Therefore, the common reversed phase column can be used for the HPLC analysis of highly hydrophilic melamine. The detection limit obtained by the proposed method was lower than 0.40 μg/L. This is the first time that HPLC with fluorescence detection was applied to the analysis of melamine. The proposed method was successfully applied to the analysis of melamine leached from tableware. The results indicated that the leaching of melamine from tableware was obvious when hot water or milk was added.<br>• HPLC with fluorescence detection was applied to the analysis of melamine for the first time. • Reversed phase HPLC analysis of melamine was achieved with no ion-pair reagents needed. • HPLC sensitivity was greatly enhanced through derivatization.