A sensitive iron assay was developed for measuring non-heme and loosely bou
nd iron in regions of rat brain. The method is based on the salicylate trap
ping of hydroxyl radicals generated from ascorbate-driven redox cycling of
Fe3+-EDTA. This assay has high sensitivity (about 20 nM) because of amplifi
cation obtained with redoxcycling and fluorescent detection of the salicyla
te hydroxylation product, 2,5-dihydroxybenzoate. The assay detects iron as
Fe2+ and Fe3+ combined. Values of non-heme and loosely bound iron are given
for three areas of cortex, caudate, hippocampus, thalamus and brainstem of
the rat brain.