Objective: To determine if children aged 1-6 years from non-Western immigrant families have lower serum 25-hydroxyvitamin D (25(OH)D) levels than children from Western-born families and examine which factors influence this relationship. Design: Cross-sectional study. Setting: Toronto, Canada. Subjects: Healthy children (n 1540) recruited through the TARGet Kids! practicebased research network. Serum 25(OH)D concentrations of non-Western immigrants were compared with those of children from Western-born families. Children from non-Western immigrant families were defined as those born, or their parents were born, outside a Western country. Univariate and multiple linear regression analyses were used to identify factors which might influence this relationship. Results: Median age was 36 months, 51 % were male, 86 % had 'light' skin pigmentation, 55 % took vitamin D supplements, mean cow's milk intake was 1?8 cups/d and 27 % were non-Western immigrants. Median serum 25(OH)D concentration was 83 nmol/l, with 5 % having 25(OH)D , 50 nmol/l. Univariable analysis revealed that non-Western immigrant children had serum 25(OH)D lower by 4 (95 % CI 1?3, 8?0) nmol/l (P 5 0?006) and increased odds of 25(OH)D , 50 nmol/l (OR 5 1?9; 95 % CI 1?3, 2?9). After adjustment for known vitamin D determinants the observed difference attenuated to 0?04 (95 % CI 24?8, 4?8) nmol/l (P 5 0?99), with higher cow's milk intake (P , 0?0001), vitamin D supplementation (P , 0?0001), summer season (P 5 0?008) and increased age (P 5 0?04) being statistically significant covariates. Vitamin D supplementation was the strongest explanatory factor of the observed difference. Conclusions: There is an association between non-Western immigration and lower 25(OH)D in early childhood. This difference appears related to known vitamin D determinants, primarily vitamin D supplementation, representing opportunities for intervention.