Speciation of iodine in commercially available commonly consumed seaweed samples was performed using a multidimensional chromatographic approach coupled with inductively coupled plasma mass spectrometry (ICP-MS) for element specific detection. Analysis of alkaline extract (0.1 mol l⁻¹ NaOH) by size-exclusion chromatography coupled to ICP-MS (0.03 mol l⁻¹ Tris-HCl, pH 8.0) indicated the association of iodine with both high as well as low molecular weight fractions in Wakame, while in case of Kombu, only low molecular weight iodine species were found. Likely association of iodine with protein as well as polyphenolic species was indicated in the case of Wakame. Anion-exchange chromatography coupled to ICP-MS (0.005 mol l⁻¹ NaOH) confirmed that the most predominant inorganic iodine species present in both type of seaweeds is iodide. Protein bound iodinated species were hydrolyzed by enzymatic digestion using Proteinase K. Analysis of the hydrolysate using reversed-phase HPLC-ICP-MS (0.01 mol l⁻¹ Tris-HCl pH 7.3 ∶ 0.01 mol l⁻¹ Tris-HCl pH 7.3 and 50% MeOH) revealed the presence of monoiodotyrosine and di-iodotyrosine in Wakame, which was later identified by matching the chromatographic retention time with the retention time of commercially available standards.