Breastfed infants depend on sufficient maternal iodine intake for optimal growth and neurological development. Despite this, few studies have assessed iodine concentrations in human milk and there is currently no published data on iodine status among lactating women in Norway. The aim of this study was to assess iodine concentrations in breast milk (BMIC) in lactating women and estimate iodine intake. Five Mother and Child Health Centres in Oslo were randomly selected during 2016, and 175 lactating women between 2nd and 28th weeks postpartum participated. Each of the women provided four breastmilk samples which were pooled and analysed for iodine concentrations. Participants also provided information on iodine intake from food and supplements covering the last 24 h and the habitual iodine intake (food frequency questionnaire). The median (p25, p75 percentiles) BMIC was 68 (45, 98) µg/L and 76% had BMIC <100 µg/L. Only 19% had taken an iodine-containing supplement during the last 24 h. The median 24 h iodine intake from food (p25, p75) was 121 (82, 162) µg/day and the total intake (food and supplements) was 134 (95, 222) µg/day. The majority of lactating women had suboptimal BMIC and inadequate intake of iodine from food and supplements.