The metabolic fate of esfenvalerate (1), 14C-labeled at the chlorophenyl or phenoxyphenyl ring, in tomato plants was investigated by spraying it three times at 15 g/ha. The overall metabolic trend of 1 was similar in foliage and fruit. The applied 1 gradually penetrated into the foliage/fruit, and approximately 30% of the total radioactive residue (TRR) distributed within the plant. The applied radioactivity remained mostly intact on the plant surface, while its degradation proceeded via ester cleavage to produce two corresponding acids derived from the chlorophenyl and phenoxyphenyl moieties, followed by saccharide conjugation at the inner tissues (each <5%TRR). While 1 retained its optical configuration (2S,αS) on the plant surface and in the fruit, a very slight isomerization at the α-cyanobenzyl carbon occurred to form a (2S,αR) isomer in the foliage (≤1%TRR). The isomerization at another asymmetric carbon C2 in the isovaleric acid moiety did not proceed on/in the plant for 1 or its metabolite.