Abstract Following a request from the European Commission, the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) derives Dietary Reference Values (DRVs) for choline. In this Opinion, the Panel considers dietary choline including choline compounds (e.g. glycerophosphocholine, phosphocholine, phosphatidylcholine, sphingomyelin). The Panel considers that none of the biomarkers of choline intake or status is suitable to derive DRVs for choline. The Panel considers that Average Requirements and Population Reference Intakes for choline cannot be derived for adults, infants and children, and therefore defines Adequate Intakes (AIs). For all adults, the Panel sets an AI at 400 mg/day based on the average observed choline intake in healthy populations in the European Union and in consideration of the amounts of choline needed to replete about 70% of depleted subjects who showed signs of organ dysfunction in a depletion/repletion study. For all infants aged 7–11 months, the Panel proposes an AI of 160 mg/day, based on upwards extrapolation from the estimated choline intake of exclusively breast‐fed infants from birth to 6 months. For all children aged 1–17 years, the Panel proposes AIs, based on downwards extrapolation from the adult AI, applying growth factors. These AIs range from 140 mg/day (1–3 years) to 400 mg/day (15–17 years). For pregnant women, the Panel derives an AI of 480 mg/day, calculated by extrapolation from the AI for non‐pregnant women and the mean gestational increase in body weight. For lactating women, the amount of choline secreted per day in human milk during the first 6 months of exclusive breastfeeding (120 mg/day) is added to the AI for non‐lactating women and an AI of 520 mg/day is set.