The search for alternative methods aiming to the control of agricultural pests that cause less environmental impact has been a recurring theme in several studies. For this purpose, the factors influencing the extraction and isolation of the trypsin inhibitor found in castor-bean (Ricinus communis L.) cake and seeds were studied, and its effect on the development of fall armyworm Spodoptera frugiperda (Smith, 1797) was evaluated. Therefore, the type and proportion of the solvent (w/v), extraction period and oil content were evaluated. The castor-bean cake extract with oil content of 20% in water at a ratio 1:40 (w/v) for 30 min showed the greatest trypsin inhibition, with 21.23 mUTI g-1 seed. The isolation of the trypsin inhibitor contained in the extract was performed by flash chromatography, with inhibition peaks of up to 8906.12 mUTI g-1 fraction, as well as inferring that the best eluent was ethyl acetate with a small percentage of ethanol (< 30%). In order to evaluate the effect of the inhibitor on the development of this insect, the aqueous extract of castor bean cake with oil content of 20% was incorporated into the diet that was provided to the caterpillars at a concentration of 4000 ppm. There was reduction in the oviposition period and an increase in the concentration of proteins in the feces, with 20.55 ± 3.73 d and 177.70 ± 16.90 &#956;mol g-1, respectively. The increase in protein excretion indicates that the trypsin inhibitor was able to reduce the absorption of essential amino acid residues by the insect without, however, impairing the development of S. frugiperda.