Spodoptera litura (Fabricius) is economically important in many countries. Many biopesticides have been developed and studied to control this pest species; however, few natural products from marine organisms have been studied for their pesticidal efficiency. This study compared the acute contact toxicity of solvent extracted from the viscera of Holothuria atra and isolated triterpene glycoside compounds on third-instar S. litura. Our results showed that crude methanolic extracts were the most toxic to the larvae (LC₅₀ ~ 10.37 mg/ml) compared with other crude extracts and contained the bioactive compound 3,4-dihydroxybenzoic acid (LC50 ~ 6.01 mg/ml). All crude extracts and isolated triterpene glycoside compounds inhibited carboxylesterase activity. The efficiency and effectiveness of H. atra extractions and triterpene glycoside compounds against S. litura indicate their potential use in integrated pest management programs and the overall value of marine species as natural sources of insecticidal compounds.