Background: Deficiency in Vitamin D is a global health issue, in the recent decade, there has been substantial proof that low levels of Vitamin D can lead to thyroid disorders. The present study aims to analyze the relationship between hypothyroidism and the level of Vitamin D, as well as to analyze the relationship between Vitamin D level with thyroid antibodies (TPO-Abs and TG-Abs). Methods: Forty patients diagnosed as hypothyroidism which compared with twenty healthy control groups with matching in sex and age between studied groups. The blood sample was collected from both groups to estimate the level of vitamin D, T3, T4, TSH hormones and the existence of thyroid autoantibodies. Results: Patients with hypothyroidism showed a significantly lower mean of Vitamin D level compared with the healthy control group (p=0.0001). Half (20/40) of the hypothyroid group tested positive for TPO-Ab, (15/40) were positive for TG-Ab, and (14/40) of hypothyroidism patients have positive results for Anti-Thyroid Peroxidase and AntiThyroglobulin antibodies with highly statistical significant between hypothyroid group according to the presences of autoantibodies. As for the interaction of vitamin D levels with autoantibodies in patients with hypothyroidism, we observed that levels of Vitamin D were correlated with neither TPO antibodies (p=0.292) nor TG antibodies (p=0.108). Conclusion: The levels of vitamin D were significantly lower in patients with hypothyroidism relative to healthy control. Vitamin D does not have a strong association with the titers of thyroid antibodies.