This study is the first to evaluate nuclear magnetic resonance (NMR) -based metabolomics in cows with displaced abomasum (DA), which is an internationally recognized problem in dairy cows. Some biochemical parameters have been used for monitoring DA. However, to date, few data have been available on the blood metabolomic profile of dairy cows. Forty Holstein multiparous cows with DA (30 left, 10 right) and 10 clinically healthy Holstein multiparous cows were the subjects of the study. All the animals had similar dry matter intake. An NMR-based metabolomics approach and hematological and biochemical analyses were performed. Some changes in biochemical parameters were observed between the groups. Among the cows with left displaced abomasum (LDA), 10 were associated with concomitant ketosis. Disease periods between the 2 DA groups were different. The metabolites identified and quantified by NMR analysis were valine, 3 beta-hydroxybutyrate (BHB), alanine, glutamine, glutamate, and succinate. The last of these was significantly decreased in cows with right displaced abomasum (RDA). Glutamine, glutamate, and 3 BHB levels were significantly different between DA groups. There was a positive correlation between BHB and valine, glutamine, and glutamate in the LDA group. Overall, this work suggests that the additional information obtained by NMR-based metabolomics evaluation may contribute to assessing the metabolic status of cows with DA.