Little is known about early development of the near-threatened bonefish (Albula vulpes), a member of superorder Elopomorpha. Members of Elopomorpha are partially defined by their synapomorphic leptocephalus larval stage, for which the nutritional requirements are not well understood. Characterizing the nutritional profile, including major nutrients (such as lipids) used for energetic processes, can help to gain a better understanding of the nutritional requirements for leptocephalus larvae. A total of 24 settlement stage A. vulpes leptocephalus larvae were collected at Long Caye Island, Belize. Samples were used to determine various biochemical characteristics including lipid class, fatty acid and glycosaminoglycan compositions. Each of these biochemical components plays a role in early developmental processes such as cellular membrane formation and is crucial for healthy development. Biochemical characteristics of settlement stage A. vulpes leptocephalus are presented in this study for the first time. The dominant lipid classes and fatty acids detected in these samples were consistent with prior studies using closely related species like the Japanese eel, indicating possible similarities in diets at this stage. In the future, similar analyses can be applied to other species that share the leptocephalus life stage to determine if nutritional requirements at this stage of development are unique to this species. The findings in this study will also help to facilitate the establishment of adequate aquaculture systems for captive bonefish, ultimately leading to improved management strategies for wild bonefish habitats.