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Purpose: Gestational diabetes that occurs during pregnancy leads to changes in lipid metabolism. It was aimed to determine the levels of LDL subfractions, betatrophin, glycosylphosphatidylinositol-anchored high-density lipoprotein binding protein 1 (GPIHBP1), and paraoxonase activity in patients with gestational diabetes mellitus (GDM), and compare with the healthy pregnant controls.Methods: This prospective case-control study included 41 women separated into two groups as GDM, and the pregnant controls. Serum levels of betatrophin and GPIHBP1 were measured by ELISA kits. LDL subfraction analysis was performed electrophoretically by using the Lipoprint LDL subfraction kit. Serum paraoxonase activity was determined using photometric paraoxonase assay kit.Results: Serum betatrophin, and GPIHBP1 levels were found to be significantly higher in patients with GDM compared to the pregnant controls (p < 0.001). Higher levels of LDL6 subfraction and smaller mean LDL size was also found in GDM patients. No significant difference was found in PON1 activity.Conclusion: Betatrophin, and GPIHBP1 may have a role in the pathogenesis of gestational diabetes mellitus. These biomarkers should be assessed in the postpartum period with long-term clinical outcomes such as atherosclerosis and cardiovascular diseases.