Abstract: Seabirds are considered as effective sentinels of environmental marine contamination and their feathers are extensively used as non-lethal samples for contaminant biomonitoring. This tissue represents the main route for mercury (Hg) elimination in seabirds and contains predominantly methylmercury (MeHg). In this work, we developed a robust analytical technique for precise and accurate simultaneous quantification of MeHg, inorganic Hg (iHg) and consequently total Hg (THg), in feathers by gas-chromatography (GC)-ICPMS analyses using species-specific isotope dilution technique. An optimisation of the extraction method was carried out by testing different extraction systems, reagents and spiking procedures using an internal reference feather sample. The procedure was validated for MeHg and THg concentrations with a human hair certified reference material. Microwave nitric acid extraction with spike addition before the extraction provided the best recovery and was chosen as the most appropriate species simultaneous extraction method (SSE). An additional assessment was performed by comparison of our developed extraction method and a MeHg specific extraction technique (MSE) classically used for Hg speciation studies on feathers. The developed method was applied to feather samples from a large number of seabirds from the Southern Ocean (penguins, albatrosses, petrels and skuas) to investigate the variability of Hg speciation across a large range of Hg exposure conditions and concentrations. In all cases, MeHg accounted for > 90% of THg, thus verifying the predominance of organic Hg over iHg in feathers.