Vitamin D deficiency is a common health problem among Libyan women. Age, gender, inadequate exposure to sunlight, and obesity are common risk factors for this issue. In this study, we randomly examined 40 Libyan women (age: 17.58±10.45, mean ±SD, range: 15 years to 65 years). Blood samples were taken from each participant and directly centrifuged and processed on Cobas 411 Automatic Electrochemil-uminescence Immunoassay Analyzer. Vitamin D levels were measured in ng/ml and the mean serum value was calculated for the total. Body weight and height for each participant were taken to calculate the body mass index. The findings revealed that all participants are suffering from vitamin D deficiency with lesser than 30 ng/ml serum levels. Female participants aged between 55 and 65 years tend to have a less degree of vitamin D deficiency as compared to the other age subgroups with a mean level of 27 ng/ml of vitamin D. This study revealed the association between vitamin D levels and obesity as measured by body mass index, in contrast, it found that overweight participants have higher vitamin D levels as compared to other groups of body mass index.