Vitamin D is an essential nutrient that plays a crucial role in calcium homeostasis and bone health. Recent research suggests that vitamin D may also have an impact on lipid metabolism, specifically the level of circulating lipids in the blood. We aim to investigate it role among healthy participate. We conducted a cross-sectional study of 15,600 patients who were referred to the laboratories of university hospitals. We measured the serum levels of Vitamin D as well as triglycerides, total cholesterol, LDL, and HDL using ELISA. We found that the mean serum level of Vitamin D was 40.31 ± 20.79 ng/mL. Of the participants, 16.7% had a serum level of Vitamin D less than 20 ng/mL, 57.7% had a level between 21 and 40 ng/mL, and 13.5% had a level between 41 and 60 ng/mL. Additionally, 12.2% had a level greater than 60 ng/mL. We performed a one-way analysis of variance and found that as the serum level of Vitamin D increased, the mean LDL level decreased significantly. Our study provides evidence of a significant relationship between serum levels of Vitamin D and LDL levels in patients. The findings suggest that vitamin D status may play a role in regulating lipid metabolism and may have implications for the prevention and treatment of cardiovascular disease. Further research is needed to elucidate the underlying mechanisms of this relationship and to determine optimal levels of vitamin D intake for maintaining lipid profiles.