To better satisfy the application of rapid detection methods in the detection of As(V) in complex food substrates, we developed an "off-on" fluorescence assay to detect As(V) based on the competition between the electron transfer effect of nitrogen-doped carbon dots (N-CDs)/Fe3+ and the complexation reaction of As(V)/Fe3+, using N-CDs/Fe3+ as a fluorescence probe. Solid-phase extraction (SPE) was used to eliminate matrix interference during sample pretreatment. The detection limit was 7.6 ng g-1, with a linear range of 10-100 ng g-1. The method was further used to determine As(V) in different seafood products including snapper, shrimp, clams, and kelp. At the same time, the recovery of the method was validated by high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP/MS), indicating that the developed method had good recoveries from 86% to 117% and met the needs for accurate determination of As(V). This approach has shown excellent application potential in the field of As(V) detection in various seafood products.