Vitamin D insufficiency has been suggested as a risk factor for several metabolic disorders. The objective of the study was to investigate the association between serum 25 hydroxyvitamin D [25(OH)D] and metabolic health markers of Brazilian individuals with normal-weight, overweight or obesity. We hypothesized that serum 25(OH)D would be inversely associated with glycemic, lipid and inflammatory markers indicative of metabolic abnormality. Data of 511 individuals (33-79 years), recruited from a longitudinal investigation (Pró-Saúde Study), were analyzed cross-sectionally. Anthropometric, biochemical, body composition, socio-demographic and lifestyle data were collected. Based on body mass index (BMI; normal weight, overweight, obesity) and metabolic health (metabolically healthy (MH) and metabolically unhealthy (MU)) categories, the participants were classified into 6 phenotypes. Individuals having zero components of the metabolic syndrome were considered as "MH". MH obesity was frequent in 2.0% of the participants and 56.0% exhibited vitamin D insufficiency (<20 ng/mL). In the subgroups of the same BMI category, there were no significant differences in 25(OH)D concentrations between individuals classified as MH and MU. After adjustments (including %body fat and BMI), an inverse association was observed between 25(OH)D and visceral adipose tissue (B = -6.46, 95% confidence interval, CI: -12.87, -0.04), leptin (B = -0.09, 95% confidence interval, CI: -0.14, -0.03), insulin (B = -0.21, 95%CI: -0.34, -0.07), HOMA-IR (B = -0.06, 95%CI: -0.10, -0.02), triglycerides (B = -2.44, 95%CI: -3.66, -1.22), and TNF-α (B = -0.12, 95%CI: -0.24, -0.005) only in MU individuals. Our results indicate that the association of 25(OH)D concentrations with a favorable biochemical profile (glycemic, lipidic and inflammatory) seems to depend on the individual's overall metabolic health, suggesting more benefits from higher serum vitamin D in MU individuals, regardless of their adiposity.