In nonruminants, low-density lipoprotein (LDL) C16:0 and C24:0 ceramide mediates insulin resistance. In dairy cattle, we have established that ceramide accumulates in plasma, liver, and skeletal muscle during the peripartum. Moreover, hyperlipidemia and steatosis promotes ceramide synthesis in cows. Liver-derived ceramide may contribute to impaired insulin action during the peripartum. Therefore, our objective was to characterize changes in lipoprotein ceramide levels in periparturient dairy cows. In a non-randomized controlled trial, 25 pregnant, multiparous lean (BCS 3.0 ± 0.2) or overconditioned (OVER; BCS 3.9 ± 0.3) Holstein cows were enrolled 28 d before expected parturition. Blood samples were routinely collected. Liver biopsies and the hyperinsulinemic-euglycemic clamp were performed 2 wk before and after parturition. Serum lipoproteins were isolated using fast protein liquid chromatography. Sphingolipids were quantified using mass spectrometry. Data were analyzed using a mixed model with repeated measures. Very low density lipoprotein (VLDL) total ceramide levels declined postpartum (730 vs. 1207 ng/mL; P < 0.01), whereas LDL total ceramide increased postpartum (1353 vs. 913 ng/mL; P < 0.01). C24:0 ceramide represented the most abundant ceramide in VLDL and LDL. OVER had lower VLDL C16:0 ceramide, relative to lean (71 vs. 86 ng/mL; P < 0.01). LDL C16:0 and C24:0 ceramide increased by d 10 postpartum, relative to d −12 (~50% increase; P < 0.01). Prepartum LDL C24:0 ceramide levels were greater in OVER (659 vs. 435 ng/mL; P < 0.05), relative to lean. The ratios of LDL C24:0 ceramide to sphingomyelin were greater in OVER (P < 0.05). LDL C24:0 ceramide were positively correlated with serum fatty acids (P < 0.05). Prepartum LDL total and C24:0 ceramide were positively correlated with liver lipid content (P < 0.05). Lastly, postpartum C16:0 LDL ceramide levels were inversely related to systemic insulin action (clamp index; P = 0.05). We conclude that LDL ceramide accrual develops with steatosis, hyperlipidemia, and insulin antagonism in peripartal cows.