Objectives: This study explored changes in serum lipid profiles among healthy Sudanese women during the second and third trimesters of pregnancy, with a focus on total cholesterol, triglycerides, LDL, and HDL levels. Method: A cross-sectional analysis was carried out involving 60 pregnant women (30 in the second trimester and 30 in the third trimester), compared with 30 healthy, non-pregnant women of similar age as a control group. Serum lipid levels were assessed using a BTS-Biosystem analyzer, and the data were analyzed using SPSS software. Findings: The study found significant increases in total cholesterol (second trimester: 193.07±38.4 mg/dL; third trimester: 219.6±44.6 mg/dL), triglycerides (second trimester: 135.4±45.9 mg/dL; third trimester: 150.1±61.6 mg/dL), and LDL cholesterol (second trimester: 110.6±29.2 mg/dL; third trimester: 126.07±36.1 mg/dL) among pregnant women compared to the control group (p < 0.05). HDL cholesterol also showed a notable increase in the third trimester (63.5±13.3 mg/dL) compared to controls. Furthermore, lipid levels were found to be significantly higher in the third trimester than in the second trimester. Novelty: This study is the first to investigate lipid profile changes specific to pregnancy trimesters in Sudanese women. It highlights trimester-related increases in lipid levels, providing essential insights for the management of hyperlipidemia and pregnancy-related complications.