Background: Non-alcoholic fatty liver disease (NAFLD) is a growing global health concern, characterized by excessive fat accumulation in the liver. This study compares the association between serum 25-hydroxy vitamin D levels in NAFLD patients with normal healthy individuals.Material and Method: The present study was conducted on 80 subjects, including 60 cases of ultrasonographically diagnosed NAFLD and 20 healthy age and gender-matched subjects as controls. In our research, 25-hydroxyvitamin D and serum triglyceride levels were evaluated in both participants, and the results were subjected to statistical analysis. Result: The mean level of 25(OH) vitamin D was 12.2 ±6.8 ng/ml in the study group which was lower compared to the mean value of 30.1 ± 17.9 ng/ml in the control group. The p-value was <0.01 which was statistically significant. vitamin D deficiency was associated with a significantly high risk of NAFLD and more prevalent in females. Conclusion: NAFLD is a hepatic manifestation of metabolic syndrome and a risk factor for type 2 diabetes mellitus, dyslipidemia, and hypertension. In the present study, we observe that serum 25 hydroxyvitamin D levels in NAFLD patients were significantly lower than in non-NAFLD patients.