The biological effects of oxidized LDL (oxLDL) may contribute to initiation and progression of the atherosclerotic process, and the association between cardiovascular disease and oxidation of LDL has been largely demonstrated. The objectives of this study were to establish the reference values of oxidative stress biomarkers in a young healthy Spanish population to determine the concentration of oxLDL and its relationship with lipid profile and with these biomarkers. oxLDL, F(2)-isoprostanes, protein carbonyls (PC), and 8-hydroxy-2'-deoxyguanosine (8-OHdG) were determinate by ELISA in 72 healthy subjects. Antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR) were carried out on a Hitachi 912 analyzer; lipid profile were assayed using automated systems (Cobas 711, Roche Diagnostics). All statistics were analyzed by using SPSS for Windows 15.0. SPSS Inc, Chicago, IL, USA. (Normal mean reference values): oxLDL (63.23 +/- 16.23 U/L), (Male/Female 68.06 +/- 17.69/58.39 +/- 13.6 U/L), F(2)-isoprostanes (2.26 +/- 0.9 microg/g creatinine), PC (0.34 +/- 0.15 nmol/mg), 8-OHdG (23.27 +/- 10.58 ng/ml), SOD (931.97 +/- 271.09 U/g Hb), GR (46.56 +/- 11.68 U/L), GPx (27.58 +/- 6.89 U/gHb (Male/Female 25.91 +/- 5.03/29.2 +/- 8.07 U/L)). OxLDL (63.23 U/L) was significantly (p < 0.05) positively correlated with BMI (22.53 Kg/m(2)), total cholesterol (175.79 mg/dl), triglycerides (87.58 mg/dl), LDL cholesterol (96.25 mg/dl), and uric acid (4.78 mg/dl), while negatively correlated with HDL-cholesterol (62.25 mg/dl). We have found different correlation between oxLDL and isoprostanes by gender with the rest of parameters. Normal reference values have been found significantly different for oxLDL and GPx by gender. Oxidized LDL is correlated with lipid profile but not with the oxidative stress biomarkers. Urinary isoprostanes are positively correlated with triglycerides and negatively with GR and GPx.