Black soldier fly (BSF, Hermetia illucens) represents a valuable source of biomolecules and it also constitutes an economic way to valorise residual biomasses. BSF prepupae contain high amounts of lipids (37% DM basis). The present investigation aimed at studying the composition of BSF lipids and the effect of killing/storage on their quality. The main fatty acid was lauric acid, sterols were represented primarily by beta-sitosterol and campesterol. Global fatty acid and sterol profiles, determined by GC-MS, were only slightly affected by the killing procedure, while lipid classes distribution, determined by 1 H NMR, strongly changed. Prepupae killed by freezing showed a drastic reduction of acylglycerols during storage and a relevant release of free fatty acids, likely due to activation of lipases. On the contrary, prepupae killed by blanching have a stable lipid fraction constituted mainly by triacylglycerols. Therefore, killing procedure strongly influences BSF oil composition and the potential applications.