The WHO recommends exclusive breastfeeding for the first 6 mo of life to promote optimal infant health and development. Understanding the micro- and macronutrient concentrations of human milk and how each nutrient fluctuates with lactational stage, maternal factors, and supplementation is imperative for supporting good breastfeeding practices. Where maternal undernutrition compromises human milk quality, a thorough awareness of the effectiveness of interventions can direct efforts to achieve both maternal and infant nutrient sufficiency. This review of current knowledge covers trends in nutrient concentrations over the course of lactation and describes the influence of maternal intake, status, supplementation, and other factors on human milk concentrations of each nutrient.