Background: Considering the role of Vitamin D in cardiovascular disease (CVD) and the relationship between coronary artery calcification (CAC) and CVD, we aimed to investigate the association between the serum level of Vitamin D and CAC. Materials and Methods: This was a cross-sectional study on 67 consecutive patients who were referred for performing computed tomography angiography. We used Spearman correlation to evaluate the relationship between Vitamin D and CAC and then linear regressions to control for demographics and vascular risk factors. Results: There was no association between CAC and Vitamin D levels (Spearman coefficient = −0.03, P = 0.805). After controlling for age, sex, hypertension, hyperlipidemia, diabetes mellitus, and smoking, there was still no association between Vitamin D and CAC score (estimate = 0.001, S. E. = 0.020, P = 0.942). Conclusion: We did not find the association between the serum level of Vitamin D and coronary artery calcification.