Background: In recent years, it is well-known that vitamin D deficiency has many complications, especially in elderly people and especially in Iran due to existence of multiple risk factors. So, awareness of vitamin D average and identify risk factors can be helpful to find ways to improve life in elderly. Methods: This cross-sectional study was conducted in Isfahan, Iran, during 2011. Our cases were 180 women with the age of more than 60 years referred to 2 laboratories in Isfahan. The 25(OH)D serum level were measured by ELISA method and then, final statistical evaluations were analyzed. Findings: The mean vitamin D concentration was 141.55 ± 70.13 nm/l and the minimum and maximum serum levels of vitamin D were 36.9 and 365.8 nm/l, respectively. The ANOVA test showed that the mean vitamin D level had a significant difference according to age group (P = 0.043) and body mass index (BMI) (P = 0.023). The mean vitamin D level was significantly higher in women using vitamin D supplements (P = 0.003). Conclusion: According to this study, due to prevalence of vitamin D deficiency in Iran, it is necessary to identify its risk factors and prevent it in elderly people. Keywords: Vitamin D, Deficiency, 25(OH)D, Elderly, Complications