In the perennial strawberry production system, removal of the harvested crop represents a loss of nitrogen (N) that may be influenced by cultivar. Eight strawberry (Fragaria ×ananassa Duch.) cultivars and eight numbered selections grown in advanced matted row culture were compared over three seasons for removal of N in the harvested crop. Replicated plots were established in 1999, 2000, and 2001 and fruited the following year. `Allstar', `Cavendish', `Earliglow', `Honeoye', `Jewel', `Northeaster', `Ovation', and `Latestar' and selections B37, B51, B244-89, B683, B753, B781, B793, and B817 were compared for yield and fruit N concentration. Harvest removal of N (HRN) was calculated from total season yield and fruit N concentration at peak harvest. There were significant differences in HRN among genotypes, ranging from 1.80 to 2.96 g N per meter of row for numbered selections B781 and B37, respectively. Among cultivars, HRN ranged from 2.01 to 3.56 g·m–1 for `Ovation' and `Jewel', respectively. The amount of HRN was largely determined by yield, however, there were also significant genotype differences in fruit N concentration, ranging from 0.608 to 0.938 mg N per gram fresh weight for B244-89 and `Jewel', respectively. These differences indicate that N losses in the harvested crop are genotype dependent.