1. During pathological states of iron-overload or oxidant stress, low-molecular-mass iron can become available within extracellular fluids.
2. This iron would be converted to the ferrous state were it not for the protective anti-oxidant protein caeruloplasmin.
3. The ferrous-ion-oxidizing activity of caeruloplasmin rapidly converts ferrous ions back to the less reactive ferric state so that they can bind to available binding sites on transferrin.
4. Cerebrospinal fluids, however, often appear to contain low-molecular-mass iron, high levels of ascorbate and low levels of ferroxidase activity with little or no iron-binding capacity.
5. When iron ions are present in cerebrospinal fluid they are therefore likely to be in the ferrous state.
6. The development and application of an assay to speciate and measure ferrous ions in simple aqueous solution and their redox cycling activity in biological fluids is described.