Here we report the isolation of lolitrem A (4) from perennial ryegrass seed, its structure elucidation using mass spectrometry and one- and two-dimensional lH and 13C NMR techniques, and its tremorgenic activity in a standard mouse bioassay. We also report assignment of the proton and carbon resonances for the methyl groups of lolitrem B, completing the partial assignment reported by Gallagher et al. (1984) and thereby allowing the NMR spectral data for 1 and 4 to be reliably compared.