The chemical composition of a plant essential oil can be affected by many environmental and biological factors. Understanding the role of individual constituents as well as their interactions to the overall insecticidal bioactivity is prerequisite to the use of essential oils as an alternative to conventional insecticides. In the present study, the chemical compositions of plant essential oils obtained from Thymus vulgaris (thyme) and Cymbopogon citratus (lemongrass) were analyzed by gas chromatography/mass spectrometry, and the insecticidal and cytotoxic activities of individual constituents were evaluated against third instar larvae and an ovarian cell line of Trichoplusia ni. Thymol was the most abundant compound in thyme oil and the primary active constituent in contact and cytotoxicity tests, whereas p-cymene was the most effective compound for fumigant toxicity. In lemongrass oil, citral was identified as the major active and most abundant constituent. A weak correlation between insecticidal activity and cytotoxicity was observed, indicating limitation of the latter as a screening tool for novel insecticides. Although the evaporation of thymol was enhanced at higher temperatures, its contribution to fumigant activity was limited.