Objective: The main aim of this study was to determine the association of serum 25-hydroxyvitamin D (25-OH D) levels with hormonal, clinical and metabolic profile in patients with and without Polycystic ovary syndrome (PCOS). Materials and Methods: Forty-eight normal-weight (body mass index (BMI) of 19-24.99 kg/m²) women with PCOS, 36 overweight (BMI of 25-29.9 kg/m²) women with PCOS and 56 normal-weight controls participated in the study. Blood samples were collected in the early follicular phase (between day 2 and day 5 of the menstrual cycle) at 9:00 am after an overnight fast. Circulating concentrations of 25-OH D, luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin, TSH, free testosterone, dehydroepiandrosterone sulphate (DHEA-SO4), 17-hydroxyprogesterone, sex hormonebinding globulin (SHBG), fasting insulin, fasting glucose, and lipid profile were assessed. Results: Normal weight (BMI 19-24.99 kg/m²) and overweight (BMI 25-29.99 kg/m²) women with PCOS were compared with normal-weight controls and lower 25-OH D levels were found in both PCOS groups (p<0.05 and p<0.01, respectively 25-OH D significantly negatively correlated with waist circumference (WC), waist-to-hip ratio (WHR), free testosterone and modified Ferriman-Gallwey scores, however, there was a positive correlation between 25-OH D and SHBG levels (p<0.05). Conclusion: Our findings suggest that PCOS is associated with hypovitaminosis.