Reperfusion injury is characterised by significant oxidative stress. F2-isoprostanes (F2-IsoPs) and isofurans (IsoFs), the latter preferentially produced during increased oxygen tension, are recognised markers of in-vivo oxidative stress. We aimed to determine whether increasing oxygen tension during reperfusion modified levels of plasma total IsoFs and F2-IsoPs. Forty five patients undergoing upper limb surgery were randomised to receive inspired oxygen concentrations of 30%, 50% or 80% during the last 15 minutes of surgery. Venous blood samples were taken before the change in inspired oxygen, after 10 minutes (before reperfusion) and after 15 minutes (5 minutes after reperfusion). IsoFs and F2-IsoPs were measured by gas chromatography-mass spectrometry. Venous oxygen tension and hemoglobin concentrations were also measured. Plasma IsoFs and F2-IsoP levels in the 50% and 80% O2 groups were not significantly different from the 30% O2 group. In secondary analyses, using data combining all groups, levels of IsoFs, but not F2-IsoPs, associated with higher venous oxygen tension (P=0.038). Hemoglobin negatively modified the influence of oxygen tension on levels of IsoFs (P=0.014). This study has shown for the first time, that plasma IsoFs levels associate with higher oxygen tension in a human model of reperfusion, and this effect is significantly attenuated by hemoglobin.