Preclinical and epidemiological studies showed the association between 25-hydroxyvitamin D (vitamin D) and cancer development. The aim of the study was to evaluate serum vitamin D levels of melanoma patients and compare them with other malignancies and healthy controls. A total of 87 cutaneous melanoma patients from a tertiary cancer center were included in the study. Vitamin D levels were measured with electrochemiluminescence binding assay. There were no differences between serum vitamin D levels of melanoma patients, other malignancies and healthy controls (median values: 18.8, 14.3, and 24.5 ng/ml, respectively,p > 0.05). Vitamin D deficiency (<20 ng/ml) was found in 56% of melanoma patients and 42% of healthy controls; however serum vitamin D levels in only 16% of both melanoma patients and healthy controls were sufficient (>30 ng/ml) (p > 0.05). Furthermore, no difference regarding serum vitamin D levels was found between melanoma and other malignancies. Age, gender and clinical stage were not found statistically correlated with serum vitamin D levels. Similarly, serum vitamin D level was not associated with disease duration and presence or absence of active disease. In conclusion, there was no difference between serum vitamin D levels of melanoma patients and healthy controls and it was also not associated with duration and activity of disease.