Diabetes mellitus may be associated with increased lipid peroxidation which may contribute to long-term tissue damage. To test this hypothesis, we measured hydroperoxides (ROOHs) as well as alpha-tocopherol in plasma from healthy subjects and individuals with non-insulin-dependent diabetes mellitus (NIDDM) (n = 41 and 87, respectively). ROOHs were analysed using the ferrous oxidation with xylenol orange version II (FOX2) assay in conjunction with a specific ROOH reductant, triphenylphosphine, alpha-Tocopherol was analysed by HPLC with fluorimetric detection. NIDDM patients had lower cholesterol standardised alpha-tocopherol levels as compared to control subjects (3.3 +/- 1.0 vs 5.1 +/- 2.3 (mumol/l)/(mmol/l); p < 0.0005, Mann-Whitney test): range (1.5-6.5 vs 1.9-13.0, respectively). Plasma ROOHs were substantially higher in the diabetic subjects compared to those of the control subjects (9.4 +/- 3.3 vs 4.1 +/- 2.2 mumol/l; p < 0.0005 Mann-Whitney test: range 2.7-16.8 vs 0.4-10.3, respectively). ROOH/cholesterol standardised alpha-tocopherol ratio was significantly higher in the diabetic patients compared to control subjects (3.2 +/- 1.6 vs 0.9 +/- 0.6; p < 0.0005, Mann-Whitney test: range 0.7-8.3 and 0.1-2.7, respectively). Plasma levels of ROOHs and alpha-tocopherol were similar in diabetic patients with or without complications as well as in smokers and non-smokers. The present study confirms previous findings from this laboratory that NIDDM is associated with increased oxidative stress as assessed by plasma ROOHs. Increased oxidative stress in diabetic patients appears to be related to the underlying metabolic abnormalities in diabetes, rather than to the complications of this disease. We therefore suggest that oxidative stress is an early stage in the disease pathology, which may contribute to the development of complications.