The objectives were to determine the effect of macronutrient modification on vitamin D status and if change in 25-hydroxy-vitamin D concentration influences components of metabolic syndrome in obese African American girls. Methods. Five-week intervention using reduced CHO (43% carbohydrate; 27% fat: SPEC) versus standard CHO (55% carbohydrate; 40% fat: STAN) eucaloric diet. Subjects were 28 obese African American females, aged 9–14 years. Dual energy X-ray absorptiometry and meal test were performed at baseline and five weeks. Results. Approximately 30% of girls had metabolic syndrome. Serum 25OHD increased in both groups at five weeks [STAN: 20.3 ± 1.1 to 22.4 ± 1.1 (<0.05) versus SPEC: 16.1±1.0 to 16.8±1.0 (=0.05)]. The STAN group, increased 25OHD concentration over five weeks (<0.05), which was positively related to triglycerides (<0.001) and inversely associated with total cholesterol (<0.001) and LDL (<0.001). The SPEC group, had increase in 25OHD (=0.05), which was positively related to fasting insulin (<0.001) and insulin sensitivity while inversely associated with fasting glucose (<0.05). The contribution of vitamin D status to metabolic syndrome parameters differs according to macronutrient intake. Improvement in 25OHD may improve fasting glucose, insulin sensitivity, and LDL; however, macronutrient intake warrants consideration.