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1 quantitative method for the determination of nitrofuran metabolites in seafood by liquid 2 chromatography isotope dilution tandem mass spectrometry 3 4 Abstract 19Despite the ban of nitrofurans (NFs) for use in food production in many countries in the 20 1990s, NF metabolites in food are still regularly detected during import control testing. We 21 have developed a confirmatory routine method for the detection and quantification of NF 22 metabolites in seafood using LC-MS/MS and validated the method according to the strict 23 criteria in European legislation and Codex Alimentarius. Method characteristics were found to 24 fulfill the criteria. We report for the first time a new false positive for 1-amino-2,4-25 imidazolidinedione (AHD), the metabolite of Nitrofurantoin (NFT). By using optimized 26 washing procedures, the non tissue bound false positives can be minimized. The results from 27 the validation on both lean and fatty fish and crustaceans, results from proficiency tests and 28 routine use over many years, demonstrates that the method is fit for purpose to determine NF 29 metabolites in the seafood category. 30 31