Reperfusion injury is characterized by significant oxidative stress. F(2)-isoprostanes (F(2)-IsoP's) and isofurans (IsoF's), the latter preferentially produced during increased oxygen tension, are recognized markers of in vivo oxidative stress. We aimed to determine whether increasing oxygen tension during reperfusion modified levels of plasma total IsoF's and F(2)-IsoP's. Forty-five patients undergoing upper-limb surgery were randomized to receive inspired oxygen concentrations of 30, 50, or 80% during the last 15 min of surgery. Venous blood samples were taken before the change in inspired oxygen, after 10 min (before reperfusion), and after 15 min (5 min after reperfusion). IsoF's and F(2)-IsoP's were measured by gas chromatography-mass spectrometry. Venous oxygen tension and hemoglobin concentrations were also measured. Plasma IsoF and F(2)-IsoP levels in the 50 and 80% O(2) groups were not significantly different from those of the 30% O(2) group. In secondary analyses, using data combining all groups, levels of IsoF's, but not F(2)-IsoP's, associated with higher venous oxygen tension (P=0.038). Hemoglobin negatively modified the influence of oxygen tension on levels of IsoF's (P=0.014). This study has shown, for the first time, that plasma IsoF levels associate with higher oxygen tension in a human model of reperfusion, and this effect is significantly attenuated by hemoglobin.