The importance of insects in the environment is huge, not always positive. Harmful insects are burdensome and cause large economic losses. One of the representatives of harmful insects is potato beetle (Leptinotarsa decemlineata). Researching the chemical composition of this insect can be used to more effectively protect plants against this pest. The eggs of Colorado potato beetle (Leptinotarsa decemlineata) were chosen for analysis because their composition is little known. Samples for analysis were prepared using the Folch method, and gas chromatography coupled with mass spectrometry (GC/MS) was selected for final determinations. As a result of the analyses, carboxylic acids, dicarboxylic acids, hydroxy acids, monoacylglycerols, alcohols, sterols and other organic compounds were identified in the tested samples. The highest content was determined for acids with a carbon chain C16:0–C18:0: palmitic (23 ± 0.88 µg/g), linoleic (32 ± 2 µg/g), oleic (83 ± 8.7 µg/g) and stearic (50 ± 2.2 µg/g). In the case of alcohols and sterols, the highest content was determined for docosanol (2.5 ± 0.04 μg/g) and cholesterol (23 ± 1.4 µg/g), respectively. The knowledge of the chemical composition of eggs can help in the work on effective insecticides, which will be effective only on a species of harmful insect.