Vitamin D is essential for skeletal health, calcium homeostasis and general health. The major and more stable form of vitamin D in circulation is 25-hydroxyvitamin D (25-OH-D); this is the most valuable indicator of vitamin D status. There are studies on laboratory and zoo-housed chimpanzees; however, serum vitamin D status has not been documented in chimpanzees in range countries. (1) Determine the range of circulating 25-OH-D concentrations in chimpanzees in range countries. (2) Assess the influence of age, sex, and sun exposure on 25-OH-D serum concentrations. Opportunistic blood samples were obtained from 127 clinically healthy chimpanzees. Serum 25-OH-D concentration was measured with a commercially available competitive ELISA. The median overall 25-OH-D concentration for chimpanzees in range countries was 46.24nmol/L (range: 17.10-109.23nmol/L). Males had a significantly lower concentration (40.15nmol/L) than females (49.61nmol/L), and infants (37.99nmol/L) had a significantly lower concentration than adults (46.04nmol/L). Concentrations of 25-OH-D in chimpanzees in sunnier habitats were significantly higher compared to thick tropical forest habitat. The present constitutes a large dataset of serum 25-OH-D concentrations in range country sanctuary chimpanzees and contributes to document normal ranges. Age, sex, and sun exposure influenced serum concentrations of 25-OH-D in sanctuary chimpanzees.