Pregnancy is associated with significant physiological changes, including alterations in lipid metabolism, which can have implications for both maternal and fetal health. This study aimed to evaluate serum lipid profile variations among healthy Sudanese women during the second and third trimesters of pregnancy. A total of 60 pregnant women, divided equally between the second and third trimesters, were compared with 30 non-pregnant, age-matched controls. Serum levels of total cholesterol, triglycerides, low-density lipoprotein (LDL), and high-density lipoprotein (HDL) were measured using a BTS-Biosystem analyzer. The results revealed a statistically significant increase in total cholesterol, triglycerides, LDL, and HDL levels among pregnant women compared to the control group (p < 0.01). Additionally, lipid levels were significantly higher in the third trimester than in the second trimester. A weak positive correlation was found between parity and serum levels of total cholesterol, triglycerides, and LDL, while no significant correlation was observed for HDL. These findings suggest that pregnancy, particularly in the third trimester, is associated with substantial increases in serum lipid levels, highlighting the need for monitoring lipid profiles during pregnancy to manage potential risks of hyperlipidemia and its associated complications.