Our research provides novel information concerning the insecticidal activity of Brassica alba mustard oil applied to the intestinal tract via insects’ diet against pests from the order Lepidoptera: Cydia pomonella, Dendrolimus pini, and Spodoptera exigua. The LC50 value of the oil against C. pomonella was 0.422 mg ⋅ ml–1. The LC50 of the plant oil against D. pini was 11.74 mg ⋅ ml–1. The LC50 of the botanical product against S. exigua was 11.66 mg ⋅ ml–1. The plant substance was the most active against C. pomonella in comparison with D. pini and S. exigua. The LC50 values of the oil against D. pini and S. exigua were similar. The plant oil exhibited high insecticidal activity against pests from the order Lepidoptera and may prove to be an effective biopesticide.