BACKGROUND: Pesticide uptake by a rotational crop after being used for the primary crop is a potential cause of violation against the pesticide law if the pesticide is not registered in the secondary crop. This study was conducted to investigate the plant back interval (PBI) of fluopyram for the rotational cultivation of radish. METHODS AND RESULTS: Two experimental approaches were performed the evaluation of residues in radish cultivated successively in soil 16 days after treated with fluopyram onto pepper plant (T1) and in radish cultivated in bare soil treated with fluopyram at PBI 30 and PBI 60 days (T2). A modified QuEChERS method coupled with LC/MS/MS analysis showed good linearity of matrixmatched standard calibration of fluopyram with the coefficient values of determination greater than 0.995. Recovery values at levels of 0.01, 0.05, 0.1 and 0.25 mg/kg ranged from average 84.9 to 117.6% with RSD less than 10%. Fluopyram residues in radish harvested from T1 and T2 were found as levels less than maximum residue limit. CONCLUSION: This study suggests 20~30 days as the PBI of fluopyram for the rotational cultivation of radish in the greenhouse soil treated with fluopyram used for pepper as the primary crop.