To better understand spatiotemporal variation in the trophic structure of deep-pelagic species, we examined the isotope values of particulate organic matter (POM) (isotopic baseline) and seven deep-pelagic fishes with similar diet compositions but contrasting vertical distributions across mesoscale features in the Gulf of Mexico using stable isotope and amino acid compound-specific isotope analyses. Species examined included four migratory (Benthosema suborbitale, Lepidophanes guentheri, Melamphaes simus, Sigmops elongatus) and three non-migratory zooplanktivorous fishes (Argyropelecus hemigymnus, Cyclothone obscura, Sternoptyx pseudobscura). Isotopic values of POM increased with depth, with meso- and bathypelagic samples characterized by higher δ C and δ N values relative to epipelagic samples. Despite similar diet compositions, mean δ N values of fishes spanned 3.43‰ resulting in mean trophic position estimates among species varying by 1.09 trophic levels. Interspecific differences in δ N were driven by higher δ N values in the non-migratory and deepest dwelling C. obscura (10.61‰) and lower δ N values in the migratory and shallowest dwelling L. guentheri (7.18‰) and B. suborbitale (8.11‰). Similarly, fish δ N values were correlated with depth, with the lowest values occurring in the migratory L. guentheri and B. suborbitale and highest values occurring in the non-migratory C. obscura. Our data suggest that depth-related trends in fish δ N and δ N values are driven by shallower dwelling species feeding within epipelagic food webs supported by POM with lower δ N values, while deeper dwelling, non-migratory species increasingly use food webs at depth supported by POM with elevated δ N values. Horizontal isotopic variation was observed across a large mesoscale oceanographic feature (Loop Current), with POM, three migratory, and one non-migratory species characterized by higher δ C and lower δ N values in the anticyclonic Loop Current relative to surrounding water masses. Our results demonstrate that isotopic values of POM can vary significantly over relatively small horizontal and vertical scales and that baseline variation can be conserved in the signatures of higher-order consumers. By gaining a more thorough understanding of the sources contributing to isotopic variation of deep-pelagic fishes, this paper will inform the design and interpretation of future feeding studies in the pelagic realm and advances our knowledge of deep-pelagic food web structure. 13 15 15 15 15 15 15 15 15 15 15 13 15 sourceAA sourceAA