To verify whether there is a variation in the 24-h urinary excretion of digoxin-like immunoreactivity (DLIS) in humans, we studied 18 normal adults, who collected their urines for 24-h in several portions. We then measured DLIS (by means of a sensitive RIA method), creatinine, sodium, and potassium concentrations in the urine samples. The mean urinary excretion rate for DLIS in the complete 24-h collection was 84.8 (SD 31.3) pg/min. The mean DLIS urinary excretion rate calculated for overnight collections was significantly lower than those of afternoon collections (P less than 0.01) and the 24-h collection (P less than 0.05). Significant positive correlations were found between urinary DLIS and excretion rates for creatinine (r = 0.347, P = 0.0016), Na+ (r = 0.232, P = 0.038), and K+ (r = 0.323, P = 0.003), respectively. Our data suggest that urinary excretion of DLIS is higher during "active" hours of the day, especially in the afternoon, than at rest, during the night.