Background: Prolonged decline in the ability of the kidney to regulate acid–base balance, eliminate waste products, and manage water homeostasis and entered chronic phase, toxic metabolic accumulates and erythropoietin secretion by the kidney is decreasing and causes hematological changes including decrease of HCT, MCV, RBCs and platelet counts. Hemodialysis became a practical treatment for kidney failure and is the most common method used to treat advanced and permanent kidney failure. Anemia is one of the most common complications in hemodialysis patients. Objectives: The study aimed to evaluate the prevalence of anemia among hemodialysis patients and investigate the variations of hematological parameters among anemic hemodialysis patients in the Tripoli region. Materials and Methods: The present study was conducted on 250 renal failure patients, attending Tripoli Center for dialysis and 100 normal healthy subjects. The study Ethical Committee of the medical centers and the Libyan Academy of graduate studies reviewed and approved the study design and patient consent statements were taken from each patient. Information's about the patients were recorded in a questionnaire. A blood sample of 5 ml was drawn by venous puncture from each normal healthy individual and hemodialysis patient. 2.5 ml of the blood sample was collected in K-EDTA tubes for the hematological examinations and another 2.5 ml of the blood sample was collected in a plain tubes for biochemical tests (serum urea, creatinine, and uric acid concentrations). The hematological parameters (RBCs count, Hb, HCT, MCV, MCH, MCHC, WBCs count, differential count of WBCs, and Platelets count) were determined using an automated hematology analyzer Sysmex (K- 4500) machine. The data were compared using GraphPad Prism version.9. The statistical significance of differences between groups was evaluated with the independent t-test. A P-value of <0.05 was considered significant for all statistical tests. Results: The results showed that the prevalence of anemia among hemodialysis patients was 89.8%. The degrees of anemia were 17% severe, 71.66% moderate, and 11.34% mild anemia. The types of anemia were 13.36% microcytic hypochromic, 82.59% normocytic hypochromic, and 4.05% macrocytic hypochromic anemia. RBCs, WBCs & platelets counts, Hct, MCHC, and Lymphocytes % showed a significant (P<0.01) decrease, and MCV was a significant (P<0.01) increase in the anemic hemodialysis patients compared with the healthy individuals. But, a significant (P<0.05) decrease in MCH was observed in the anemic hemodialysis patients when compared with the healthy individuals. A significant correlation was observed between RBCs and their indices with most of the hematological parameters. A significant (P<0.01) negative correlation was observed between serum urea with Hb, and RBCs count and Hct. While, a significant (P<0.01) positive correlation was recorded between uric acid with platelets count. A significant (P<0.05) positive correlation was observed between gender with platelets count, while, a significant negative correlation was recorded between gender with serum urea (P<0.01), creatinine, and uric acid, and Hb (P<0.05). A significant (P<0.01) negative correlation was observed between blood groups with serum uric acid. A significant (P<0.01) positive correlation was observed between durations of hemodialysis with RBCs count and Hb, while, a significant (P<0.05) negative correlation was recorded between durations of hemodialysis with body weight, and MCHC. Conclusion: It can be concluded that a higher prevalence of moderate, normocytic hypochromic anemia among hemodialysis patients. Also, results showed a significant variation in hematological parameters among the anemic hemodialysis patients. So, hemodialysis patients advice to examine the hematological parameters and treated from anemia if detected.