Reference values for urinary D-glucaric acid and the influence of sex, age and smoking habits were evaluated with a low-pH enzymatic method. D-Glucaric acid measured on spot urine samples from 573 healthy subjects gave mean concentrations (mumol/1) and D-glucaric acid/creatinine ratios (mmol/mol creatinine) of 56.1 (+/- 22.9) and 3.05 (+/- 0.99) for males and 53.3 (+/- 20.9) and 3.35 (+/- 0.95) for females. No difference between morning and evening was observed for urinary D-glucaric acid/1 values, but D-glucaric acid/creatinine was higher in the evening samples for both sexes. There was a negative correlation between D-glucaric acid/1 values and age in males but not in females: the decrease of D-glucaric acid concentration was, however, quantitatively very small. Smoking produced a significant increase in D-glucaric acid concentration and in the D-glucaric acid/creatinine ratio for males and also partially for females.