Wagyu crossbred steers (n = 167) were used to (1) compare the metabolome of individual animals at two distant time-points (days 196 and 432) in a feedlot (this corresponded to 272 and 36 days before slaughter); and (2) determine relationships between the metabolome and marbling, and the effect of days in the feedlot (time-points) on these relationships. 1H NMR spectroscopy followed by standard recoupling of variables analysis produced 290 features or 'peaks' from which 38 metabolites were identified. There was a positive correlation between the relative concentration (RC) at days 196 and 432 for 35 of 38 metabolites (P > 0.05). The RC of 21 metabolites mostly involved in muscle energy and glucose metabolism increased (P < 0.05) from day 196 to 432, and the RC of 13 metabolites mostly involved in lipid metabolism decreased (P < 0.05). There were 14 metabolites correlated with marbling including metabolites involved in energy and fat metabolism (glucose, propionate, 3-hydroxybutyrate, lipids). The relationship between marbling and the RC of metabolites was affected by time-point, being positive for 3-hydroxybutyrate and acetate (P < 0.05) at day 432 but not at day 196. The findings indicate that the blood metabolome in Wagyu crossbred steers changes with time in a feedlot. Notwithstanding, the metabolome has potential to predict marbling in Wagyu. The ability to predict marbling from the blood metabolome appears to be influenced by days in a feedlot and presumably the stage of development towards a mature body conformation.