Using strontium (Sr) and barium (Ba) in otoliths to determine natal origins and understand patterns of fish movements is based on the fundamental assumption that otoliths record water chemistry signals without any major alterations. Although prior studies highlighted that fish physiology can modify the water signal in otoliths, studies for freshwater fish are scarce. We exposed different groups of Atlantic salmon parr Salmo salar to different scenarios of ambient-level variations in Sr/Ca and Ba/Ca ratios and then combined otolith chemical profiles with environmental data (water chemistry and temperature), Fulton's index, and otolith growth rates to assess what factors explain/influence the elemental ratios of Sr and Ba in otoliths. Generalized additive mixed models (GAMMs) using water-based otolith composition, temperature, Fulton's index, and “individual” as explanatory variables allow to demonstrate that water chemistry alone cannot fully explain measured ratios in otoliths, except in scenarios involving significant changes in water chemistry. Other factors (physiological effects) should be accounted for reproducing short and minimal seasonal variations in water composition, considering that inter-individual variability contributes quite significantly in most scenarios.