Mild to moderate iodine deficiency is common among women of childbearing age. Data on iodine status in infants are sparse, partly due to the challenges in collecting urine. Urinary iodine concentration (UIC) is considered a good marker for recent dietary iodine intake and status in populations. The aim of this study was to investigate the reliability of iodine concentration measured in two spot-samples from the same day of diaper-retrieved infant urine and in their mothers’ breastmilk. We collected urine and breastmilk from a sample of 27 infants and 25 mothers participating in a cross-sectional study at two public healthcare clinics in Norway. The reliability of iodine concentration was assessed by calculating the intraclass correlation coefficients (ICC) and the coefficient of variation (CV). The ICC for infants’ urine was 0.64 (95% confidence interval (CI) 0.36–0.82), while the ICC for breastmilk was 0.83 (95% CI 0.65–0.92) Similarly, the intraindividual CV for UIC was 0.25 and 0.14 for breastmilk iodine concentration (BIC). Compared to standard methods of collecting urine for measuring iodine concentration, the diaper-pad collection method does not substantially affect the reliability of the measurements.