In non-ruminants, 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, the induction of 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. Twenty-five pregnant, multiparous lean (BCS 3.0 ± 0.2; n = 14) or overconditioned (OVER; BCS 3.9 ± 0.3; n = 11) Holstein cows were enrolled 28 d before expected parturition. Blood was sampled at d −10, 0, and 12, relative to calving. Liver biopsies and the hyperinsulinemic-euglycemic clamp were performed 2 wk before and after parturition. Serum lipoproteins were isolated using 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 (P < 0.01), whereas LDL total ceramide increased postpartum (P < 0.01). C24:0 ceramide represented the most abundant ceramide in VLDL and LDL. OVER had lower VLDL C16:0 ceramide (P < 0.01), relative to lean. LDL C16:0 and C24:0 ceramide increased by d 10 (P < 0.01), relative to d −12. Prepartum LDL C24:0 ceramide levels were greater in OVER (P < 0.05), relative to lean. The ratios of LDL C24:0 ceramide to sphingomyelin were greater in OVER (P < 0.05), suggesting the potential involvement of sphingomyelin hydrolysis. The levels of C24:0 ceramide were positively correlated with serum fatty acid levels (r = 0.41; P < 0.01). Prepartum LDL total and C24:0 ceramide levels were positively correlated with liver lipid content (r = ~0.50; P < 0.05). Lastly, postpartum C16:0 LDL ceramide levels were inversely related to systemic insulin action (r = −0.44; clamp index; P = 0.05). We conclude that LDL ceramide accrual develops with steatosis, hyperlipidemia, and insulin antagonism during the peripartum.