There are a lot of studies with conflicting results about the correlation between blood levels of vitamin D and lipid profile (total cholesterol, LDLc, HDLc) and the effect of treatment with vitamin D supplements on it. This study aimed to investigate the effect of vitamin D and calcium supplements on lipid profile. This study included 59 volunteers who received these supplements for prevention of osteoporosis as a prescription from orthopedic doctors between 11/2016 to 1/2018. The average of age was 49±8 years. The plasma values of TC, LDLc, HDLc were measured using enzymatic colorimetric method ( Biosystems kits ) in Tishreen hospital before treatment and after two months. The results were analyzed by T-Student test, and P value <0.05 was considered significant. The mean plasma levels of LDLc before treatment were (100.983 ±28.36) mg/dl, and after two months of treatment were (102.249±27.478) mg/dl, and the difference was not significant. The mean plasma levels of HDLc before treatment were (68.056 ±14.369) mg/dl and after two months of treatment were (62.715 ± 14.39) mg/dl, and the difference was significant. We advise to measure plasma lipid levels during treatment with these supplements depending on the changes that observed in this study.