The major objective of present study has been to investigate a correlation between an elevated level of vitamin D and serum level of uric acid in a gouty rat model created by potassium oxonate. The experiment involves examine the impact of vitamin D3 on serum uric acid in hyperuricemic rats created with potassium oxonate. Animal's study (rats) was randomly divided to four groups (n = 6). Potassium oxonate (uricase enzyme inhibitor) was injected intraperitoneally (i.p) at a dosage of 250 mg/kg to the rats of groups (B-D-Z) twice a week. Group (A) fed with food and water without treatment and served as negative control group. Group (B) orally administered with 0.5 ml of Olive Oil (vehicle) once weekly and served as positive control group. Animals of group (D) received Vitamin D3 in a dose (715 I.U./kg) orally once weekly. Group (Z) rats was treated orally with standard drug(allopurinol) (5 mg/kg) daily. Animals of each group were sacrificed and blood was collected and serum were separated for biochemical analysis of serum uric acid (SUA), serum creatinine (SCr), blood urea, AST (Aspartate transaminase), ALT (Alanine transaminase) and ALP (Alkaline phosphatase) levels, Serum levels of xanthine oxidase and Serum level of vitamin D were assay. The study's findings revealed a highly significant rise in blood uric acid and xanthine oxide in hyperuricemic rats administered vitamin D. There are non-significant differences in liver functions (ALT, AST, and AlkP) and kidney functions (Urea and CreaC) have been seen for all three groups (B, D, and Z) as compared with negative control group. This study revealed that uric acid level in the blood showed a positive correlation with vitamin D concentration.