There is a need for new, non-invasive, rapid and reliable analytical methodologies that can easily be implemented and used for authentication of cattle production systems and the meat derived from them. Easily quantifiable markers could strengthen the current tracing methods for beef authentication. This study investigated the use of a nuclear magnetic resonance-based metabolomic approach as a tool to authenticate beef on the basis of the pre-slaughter production system. Urine and muscle samples were collected from animals fed either pasture outdoor, a barley-based concentrate indoor, silage followed by pasture outdoor or silage followed by pasture outdoor with concentrate over 1 year. A metabolomic analysis was performed on urine (n 5 68) and muscle (n 5 98) samples collected from animals on the different diets. The results showed that separation according to production system was possible indicating the potential use of this approach in beef authentication. Identification of the major discriminating peaks in urine led to the identification of potential markers of production system including creatinine, glucose, hippurate, pyruvate, phenylalanine, phenylacetylglycine and three unassigned resonances.