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Objective The physiological challenges of high altitude have led to population-specific patterns of adaptation. These include alterations to child growth and reproduction, including lactation. However, while breastfeeding has been investigated, nothing is known about milk composition in high altitude adapted populations. Materials and Methods Here, we investigate milk macronutrient composition, volume, and energy in a sample of 82 Tibetans living at high and low altitude in rural villages (Nubri Valley, Nepal) and at low altitude in Kathmandu, Nepal. Milk samples were collected in the morning using hand expression, frozen, and assayed for fat, protein, and total sugars. Reproductive histories and health recalls were also collected. Results Milk fat averaged 5.2 ±2.0 g/100 mL, milk sugar 7.37 ± 0.49 g/100 mL, and milk protein 1.26 ± 0.35 g/100 mL for a mean energy density of 81.4 ± 17.4 kcal/100 mL. There were no associations between altitude of residence and milk composition; however, overall milk fat was high compared to reference populations. Within the three groups, milk fat was positively associated with infant age (B = 0.103; p