Abstract: To investigate the accumulation and elimination of four nitrofuran metabolites in muscle of grouper, we fed the groupers with 100 mg·kg^–1 of nitrofurantoin, furazolidone, furaltadone and nitrofurazone for seven days. The concentrations of semicarbazide (SEM), 3-amino-2-oxazolidone (AOZ), 5-morpholinomethyl-3-amino-2-oxazolidone (AMOZ) and 1-aminohydantoin (AHD) in muscle were determined by UPLC-MS/MS and quantified by internal standard method. The maximum concentrations of nitrofuran metabolites in grouper muscle are as follows: SEM: 1 104.10 μg·kg^–1, AOZ: 781.09 μg·kg^–1, AMOZ: 679.75 μg·kg^–1 and AHD: 437.18 μg·kg^–1. The rules of elimination for the four nitrofuran metabolites in grouper muscle are similar. The half-life time of nitrofuran metabolites in grouper muscle is as follows: SEM: 23.90 d, AHD: 23.10 d, AOZ: 22.35 d and AMOZ: 21.00 d. The eliminating rate of SEM is the lowest. By 163^rd day of elimination, the concentrations of SEM, AOZ, AMOZ and AHD in grouper muscle had still been over 0.5 μg·kg^–1. It is suggested that nitrofurans should be banned in the prevention or treatment of grouper disease.