A pivotal biochemical event in the thyroid physiology is identified unravelling a superoxide anion radical-mediated activation of iodine into an active I.- form, which could be the intermediate that is incorporated onto tyrosine. This active iodine species gives fairly stable spin-adducts with PBN that could be characterized using EPR spectroscopy. Thus, a long-lasting puzzle regarding the iodine intermediate formed before iodination of tyrosine seems to be solved.