Background:  Vitamin  D  deficiency  and  anemia   are common problems that  increased  pathophysiology  in  the  chronic kidney disease (CKD) undergoing hemodialysis (HD). This study was conducted to assess the association between vitamin D  and  anemia status  in  HD  patients.  Methods:  This  was  a  cross-sectional  study  of 65 HD patients and EPO therapy were enrolled; compared with 25 age matched healthy individuals as controls. The current study was investigated serum hepcidin, iron status  was  included  serum  ferritin and iron levels, anemia hematological indices  such  as  RBC,  Hb,  MCV, and MCH, in addition to other  clinical parameters  of  kidney  function, and C-reactive protein (CRP)  as inflammation markers were  performed in both HD patients and controls. Results: The mean patient age was (45.65±13.34) years, with dialysis vintage of median (IQR) was (12 (3- 36) month. serum hepcidin were highly significantly increase in HD patients than in the control group  (622.3±139.6,  and 465.16±80.31pg/ml, respectively p = 0.0001). serum hepcidin significantly associated with 37% anemia severity about (692.4±119.8) pg/ml more than patient with 49.2% of moderate about (605.3±117.7) pg/ml, (p=0.007) as compared with 13.8% of mild anemia groups (531.8±99.12) pg/ml, (p=0.001). negative  significant  correlation between Vt.D levels and age (r = -0.283, p=  0.022), but  no  correlated with BMI (r= 0.127, p=0.314). A  negatively  significant  between  Vt.D with dialysis vintage (r=  -0.419,  p=0.0001),  Urea  (r=  -  0.422, p=0.0001), and creatinine (r= -0.491, p=0.0001), while its strong positively significant correlated with GFR (r= 0.800, p=0.0001), Ca (r= 0.339, p=0.0001), and serum iron (r= 0.592, p=0.0001), but no with ferritin (r= -0.063, p=0.616). Moreover, vitamin D a negative correlated with blood glucose (r= -0.489, p=0.0001),  and  CRP  (r=--0.598, p=0.0001). The study concluded that Vt.D was associated with risk hemodialysis outcome and anemia severity in HD patients.