Bulk collagen δ15N analysis is widely used to investigate past diet and trophic positions, but these values average the δ15N values of the constituent amino acids. Compound specific isotope analysis of amino acids (AAs) can help elucidate the complex metabolic effects underpinning bulk δ15N values. Although trophic level effects on individual AA δ15N values have been investigated in aquatic and terrestrial invertebrate food webs, most archaeological applications involve terrestrial herbivores, hence a greater understanding of these effects between diet and consumer in this food chain is required. The North Wyke Farm Platform provided baseline nitrogen isotope information for cattle grazing on a Lolium perenne-dominated pasture. Bulk dentine δ15N values show a shift expected for a one trophic level increase, but obscure insight into the underlying metabolic processes that cause this change in value. However, determination of AA δ15N values of hydrolysable plant protein and cattle tooth dentine clarifies the trophic effect on consumer AA δ15N values. The observed trophic shift in the studied system is different from previously studied food webs, with a trophic enrichment factor, based on the δ15N values of glutamate and phenylalanine, of 4.0‰ compared to 7.6‰ commonly used in ecological and archaeological studies. This emphasises the need to understand the trophic shifts in the particular food web being investigated in order to apply isotopic investigations in archaeological contexts.