Knowledge of the subcellular behavior of toxicants, mainly metals, in living organisms is fundamental for understanding their toxicological consequences at individual and trans-generation levels and throughout the food web. However, different partitioning methodologies are available in the literature, particularly concerning the cytosolic fraction. Different cytosolic compounds, metallothionein-like proteins (MTLPs), and non-metallothionein-like proteins (non-MTLPs) could be separated by size exclusion chromatography or by heat treatment and centrifugation. Our results show changes of metal partitioning in the cytosolic fraction according to the methodologies used. After heat treatment, the majority of cytosolic cadmium (Cd) was found with MTLPs compounds and the opposite results were observed with size exclusion chromatography. Metal partitioning could also play an important role in the metal trophic transfer. Therefore, metal partitioning in Gammarus fossarum, an ecologically relevant species and good metal bioaccumulator, defining the trophically available metal fraction, is also discussed.

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