Ensuring sufficient iodine intake is a public health priority, but we lack knowledge about the status of iodine in a nationally representative population in Norway. We aimed to assess the current iodine status and intake in a Norwegian adult population. In the population-based Tromsø Study 2015-2016, 493 women and men aged 40-69 years collected 24-h urine samples and 450 participants also completed a food frequency questionnaire (FFQ). The 24-h urinary iodine concentration (UIC) was analyzed using the Sandell-Kolthoff reaction on microplates followed by colorimetric measurement. Iodine intake was estimated from the FFQ using a food and nutrient calculation system at the University of Oslo. The mean urine volume in 24 h was 1.74 L. The median daily iodine intake estimated (UIE) from 24-h UIC was 159 µg/day (133 and 174 µg/day in women and men). The median daily iodine intake estimated from FFQ was 281 µg/day (263 and 318 µg/day in women and men, respectively). Iodine intake estimated from 24-h UIC and FFQ were moderately correlated (Spearman rank correlation coefficient r = 0.39, p