Effect of ferrous iron on the generation of hydroxyl free radicals by liver microdialysis perfusion of salicylate.

1. We applied in vivo microdialysis technique to examine the effect of Fe2+, Fe3+, Cu2+ and Zn2+ on free radical-generation of rat liver. The hydroxyl free radical (.OH) reacts with salicylate and generates 2,3- and 2,5-dihydroxybenzoic acid (DHBA) which can be measured electrochemically in picomole quantity by HPLC-EC procedure. 2. The relative rates of recovery of 2,3- and 2,5-DHBA at 1 microliter/min in vitro were an average of 10.1 +/- 0.8% and 10.5 +/- 9%, respectively. 3. When the metal ion infused through the dialysis probe, Fe2+ but not Fe3+, Cu2+ and Zn2+ caused an increase in the formation of DHBA of rat liver. 4. After the Ringer solution containing 10 mumole/kg was injected into the penile vein, the levels of 2,3- and 2,5-DHBA were increased about 60% and 40%, respectively. 5. These results indicate that non-enzymatic oxidation in the extracellular fluid may play a key role in Fe2+ generation of -OH in liver. Free radical formation processes may contribute to in vivo free radical formation induced by Fe2+.