The aim of this study was to determine the relationship between the oxidative and reductive metabolic pathways of acrylamide (AA) in the nonsmoking general population. For the first time both the blood protein adducts and the urinary metabolites of AA and glycidamide (GA) were quantified in an especially designed study group with even distribution of age and gender. The hemoglobin adducts N-carbamoylethylvaline (AAVal) and N-(R,S)-2-hydroxy-2-carbamoylethylvaline (GAVal) were detected by GC-MS/MS in all blood samples with median levels of 30 and 34 pmol/g of globin, respectively. Concentrations ranged from 15 to 71 pmol/g of globin for AAVal and from 14 to 66 pmol/g of globin for GAVal. The ratio GAVal/AAVal was 0.4-2.7 (median = 1.1). The urinary metabolites were determined by LC-MS/MS. Of all urine samples examined 99% of N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA) levels and 73% of N-(R/S)-acetyl-S-(2-carbamoyl-2-hydroxyethyl)-L-cysteine (GAMA) levels were above the LOD (1.5 mu g/L). Concentrations ranged from < LOD to 229 mu g/L (median = 29 mu g/L) for AAMA and from < LOD to 85 mu g/L (median = 7 mu g/L) for GAMA. The ratio of GAMA/AAMA varied from 0.004 to 1.4 (median = 0.3). Using hemoglobin adduct levels in blood and mercapturic acid excretion in urine for calculation of daily AA intake gave practically identical values. The median daily intakes were 0.43 (0.21-1.04) mu g/kg of body weight(bw)/day using Hb adducts and 0.51 (< LOD-2.32) mu g/kg of bw/day using mercapturic acids for calculations. Children take up approximately 1.3-1.5 times more AA per kilogram of body weight than adults. The ratio GAMA/AAMA is significantly higher in the group of young children (6-10 years) with a median level of 0.5. A gender-related difference in internal exposure and metabolism was not observed.