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Semicarbazide (SEM) has been proven to extensively exist in foodstuffs due to anthropogenic factor in food processing and possesses various toxic effects on human health. Although many methods have been developed, they often require long analytical time, complex laboratory equipment, trained personnel, difficultly prepared antibodies, or relatively expensive equipment. The present study developed a new method for SEM determination by HPLC with fluorescence detection (FLD). The fluorescence reagent, 2-(11H-benzo[a]carbazol-11-yl) ethyl chloroformate (BCEC), was first used for SEM labeling. The fluorescent labeling conditions were optimized systematically. SEM can be labeled in only 10 min at 40 °C. The labeled SEM was analyzed on an eclipse XDB-C8 column in 8 min. The new method offered the low LOD of 0.4 μg/kg at a signal-to-noise ratio of 3 and also exhibited excellent reproducibility, precision, and accuracy. When applied to analyze several foodstuffs, it showed good applicability. The developed method has been proven to be simple, inexpensive, selective, sensitive, accurate, and reliable for SEM analysis in foodstuffs. Furthermore, this developed method should have a powerful potential in the analysis of SEM from many other food samples.