Seabirds accumulate significant amounts of mercury (Hg) due to their long-life span together with their medium to high trophic position in marine food webs. Hg speciation and Hg isotopic analyses of total Hg in different tissues (pectoral muscles, liver, brain, kidneys, blood and feathers) were assessed to investigate their detoxification mechanisms. Three species with contrasted ecological characteristics were studied: the Antarctic prion (zooplankton feeder), the white-chinned petrel (pelagic generalist consumer) and the southern giant petrel (scavenger on seabirds and marine mammals). The difference of mass-dependent fractionation (MDF, δ202Hg) values between livers and muscles (up to 0.94 ‰) in all three seabirds strongly suggests hepatic methylmercury (MeHg) demethylation and redistribution of the isotopically heavier fraction of MeHg towards the muscles. Similarly, higher δ202Hg values in feathers (up to 1.88 ‰) relative to muscles and higher proportion of MeHg in feathers (94-97%) than muscles (30-70%) likely indicate potential MeHg demethylation in muscle and preferential excretion of MeHg (isotopically heavier) in the growing feathers during moult. The extents of these key detoxification processes were strongly dependent on the species-specific detoxification strategies and levels of dietary MeHg exposure. We also found higher mass-independent fractionation (MIF, ∆199Hg) values in feathers relative to internal tissues, possibly due to different integration times of Hg exposure between permanently active organs and inert tissues as feathers. Hg isotope variations reported in this study show evidence of detoxification processes in seabirds and propose a powerful approach for deep investigation of the Hg metabolic processes in seabirds.