This study focused on exploring the metabolomic profiles of crossbred beef cattle with varying levels of residual feed intake (RFI), a measure of feed efficiency in beef cattle. Sixty-seven crossbred growing beef steers (BW = 277 ± 29.7 kg) were subjected to a high-forage total mixed ration for 64 days to determine their RFI phenotypes. At the end of the 64d feeding trial, beef steers were divided into two groups based on their RFI values: low (or negative)-RFI beef steers (n = 28; RFI = −1.08 ± 0.88 kg/d) and high (or positive)-RFI beef steers (n = 39; RFI = 1.21 ± 0.92 kg/d). Blood samples were collected, and plasma samples were analyzed using Nuclear Magnetic Resonance spectroscopy, resulting in the identification of 50 metabolites. The study found a distinct metabolomic signature associated with RFI status. Eight metabolites, including amino acids (tyrosine, glycine, valine, leucine, and methionine) and other compounds (dimethyl sulfone, 3-hydroxy isovaleric acid, citric acid, creatine, and L-carnitine), showed differential abundance between low- and high-RFI groups. Specifically, tyrosine, glycine, and dimethyl sulfone exhibited significant specificity and sensitivity, which produced a discriminatory model with an area under the receiver operating characteristic (ROC) curve of 0.7, making them potential markers for RFI. A logistic regression model incorporating these biomarkers effectively distinguished between high- and low-RFI steers, with a threshold cutoff point of 0.48, highlighting a distinctive metabolite profile associated with efficient nutrient utilization in low-RFI cattle. The logistic regression model, incorporating these biomarkers, holds promise for accurately categorizing RFI values, providing insights into the metabolic basis of feed efficiency in beef cattle.