An apparent digoxin-like immunoreactive substance(s) (DLIS) was evaluated in 374 pediatric patients, 0 to 72 months of age, not receiving digoxin. The relationship between DLIS presence or concentration and age, weight, gender, race, and serum creatinine was investigated. Twenty-seven percent of patients had a positive apparent DLIS concentration (greater than or equal to 0.2 ng/ml). The mean +/- SD concentration of DLIS in the positive group was 0.39 +/- 0.18 ng/ml (range 0.2 to 1.37 ng/ml). Patients in the DLIS-positive group were younger than those in the negative group (P less than 0.01). Although a greater percentage of infants younger than 6 months of age had measurable DLIS, the mean DLIS concentrations, when present, were not significantly different for all age groups (P greater than 0.05). No significant relationship was found between race or gender and DLIS. A weakly positive correlation between serum creatinine concentration and DLIS was noted (r = 0.22, P less than 0.03), but elevated serum creatinine measurements (greater than 0.6 mg/dL) did not correlate with DLIS concentration. These results support the hypothesis that the presence of DLIS is age related, but DLIS appears to be present in a much larger and older population of pediatric patients than previously reported.