PURPOSE: Urine sampling has previously been evaluated for detecting dehydration in young male athletes. The present study investigated whether urine analysis can serve as a measure of dehydration in men and women of a wide age span. METHODS: Urine sampling and body weight measurement were undertaken before and after recreational physical exercise (median time: 90 minutes) in 57 volunteers aged between 17 and 69 years (mean age: 42). Urine analysis included urine color, osmolality, specific gravity, and creatinine. RESULTS: The volunteers' body weight decreased 1.1% (mean) while they exercised. There were strong correlations between all four urinary markers of dehydration (r = 0.73 to 0.84, P < 0.001). Researchers constructed a composite dehydration index graded from 1 to 6 based on these markers. This index changed from 2.70 before exercising to 3.55 after exercising, which corresponded to dehydration of 1.0% as given by a preliminary reference curve based on seven previous studies in athletes. Men were slightly dehydrated at baseline (mean: 1.9%) compared to women (mean: 0.7%; P < 0.001), while age had no influence on the results. A final reference curve that considered both the present results and the seven previous studies was constructed in which exercise-induced weight loss (x) was predicted by the exponential equation x= 0.20 dehydration index. CONCLUSION: Urine sampling can be used to estimate weight loss due to dehydration in adults up to the age of 70 years. A robust dehydration index based on four indicators reduces the influence of confounders.