The purpose of this study is to investigate the effect of vitamin D deficiency on lipid parameters; cholesterol, triglycerides, low density lipoprotein (LDL), and high density lipoprotein (HDL). Serum level of vitamin D and the lipid parameters were determined in 127 subjects attending private community lab in eastern Amman province, using commercially available kits. Deficiency and insufficiency of vitamin D were 36.2%, 29.9%, respectively. Serum level of cholesterol, triglycerides, and LDL in deficient and insufficient vitamin D subjects were significantly increased, while HDL level was decreased, P-value <0.05. Serum levels of lipid profile and vitamin D in each of the control, insufficient , and deficient groups were all normally distributed due to the fact that P-value is greater than 0.05. Interval plot analysis and One-way ANOVA analysis revealed a significant effect of vitamin D insufficiency or deficiency on serum level of cholesterol, triglycerides, and LDL and increases the levels more than the double. Two samples T-test for every lipid parameter analysis related to subjects in the sufficient, insufficient and deficient vitamin D groups confirm our hypothesis on the effect of vitamin D deficiency on increasing the levels of cholesterol, triglycerides, and LDL and decreasing the level of HDL.