Aim: Non-alcoholic fatty liver disease (NAFLD) is a common disorder which causes serum liver enzyme elevation. Elevated homocysteine levels was demonstrated in fatty liver disease and chronic liver failure. However, enough data related with homocysteine levels in patients with NAFLD is not available. We aimed to find out whether there is an association between homocysteine levels and NAFLD. Methods: Twenty patients (14 men, 6 women) with NAFLD and 20 healthy adults (13 men, 7 women) enrolled in the study. Fasting blood samples were obtained and serum homocysteine levels were measured by fluorescence polarization immunoassay (FPIA) technology. Oral glucose tolerance test was performed and serum insulin, c-peptide, and lipoprotein levels were also measured. Results: The mean serum homocysteine levels (+/-SD) were 13.44±3.10 μmol/L and 11.62 ±1.34 μmol/L in NAFLD and the control group, respectively. Mean serum homocysteine level in the NAFLD group was significantly higher than in control group (p=0.015). Fasting blood glucose, insulin, total cholesterol and low density lipoprotein (LDL) cholesterol were all found higher than the control group. Conclusion: The serum homocysteine levels were significantly higher in patients with NAFLD than in control group. This may point out that high homocysteine levels may be associated with NAFLD.