Objectives were to investigate the effect of feeding a rumen-protected methionine (Smartamine M) or a rumen-protected choline (Reashure) source during the peripartal period on the skeletal muscle metabolome and identify the most-impacted metabolites using a high-resolution mass spectrometry-based untargeted metabolomics approach. Twenty-four multiparous Holstein cows were randomly assigned from −21 to +30 d relative to parturition to a basal control (CON) diet; CON plus rumen-protected methionine (MET) at a rate of 0.08% of DM; or CON plus rumen-protected choline (CHOL) at 60 g/d. Skeletal muscle samples from the hind-leg were harvested at −10, 7, and 20 d. Samples were analyzed using Q-Exactive MS system after LC (Liquid chromatography) separation. Data analysis was performed using the MetaboAnalyst 3.0 program. A total of 1504 metabolites were identified. Multivariate analysis (PLS-DA, and OPLS-DA) revealed clear separation of metabolomics profiles among CON, MET, and CHOL groups at −10, 7, and 20 d. Metabolites with ‘importance in projection (VIP)’ scores >1.0 and P < 0.05 were considered significantly different among treatments. The top 15 metabolites with highest VIP score were identified by molecular weight (mass error ppm <5) for comparisons among treatments at −10, 7, and 20 d, respectively. Compared with CON, lactic acid (VIP = 3.3) was lower in MET-fed cows but greater in CHOL-fed cows at −10 d. A greater skeletal muscle proline (VIP = 2.9) level also was observed in MET-fed cows but not in CHOL-fed cows compared with CON. As incomplete breakdown products of protein catabolism, although concentrations of dipeptides such as gamma-glutamyl-alanine (VIP = 2.6) and asparaginyl-valine (VIP = 2.5) were both greater in MET-fed cows compared with CON at 7 d, a lower level at 20 d of L-gamma-glutamyl-l-valine (proteolytic breakdown product of larger proteins) was observed for MET-fed cows. These preliminary analyses indicate that peripartal MET or CHOL feeding influence the skeletal muscle metabolome to different extents. The biologic significance of these alterations remains to be elucidated.