High resolution proton nuclear magnetic resonance (1H NMR) spectroscopic methods have been used to characterise tissue extracts of a series of common British invertebrate species with pollution indicator potential. These include two earthworm species Lumbricus rubellus (Hoffmeister) and Eisenia andrei (Savigny), two terrestrial isopods, Oniscus asellus (L.) and Porcellio scaber (Latreille), the diplopodous arthropod, Glomeris marginata (Villers) and a pulmonate gastropod, Arion subfuscus (Draparnaud). One and two-dimensional NMR techniques including 1H-1H homonuclear correlation and 1H J-Resolved NMR spectroscopic methods were applied to allow characterisation of the major organic components in the tissue extracts. The extracts gave characteristic low molecular weight metabolite NMR fingerprints for each species studied. Endogenous metabolites identified included glucose and trehalose, a range of free amino acids and organic acids and bases. The presence or absence of metabolites observed in the NMR spectra was examined by cluster analysis to investigate species similarity and differences in metabolite profiles. The use of Principal Component Analysis to interrogate NMR data-reduced spectra of tissue extracts allowed for distinct separation of the two morphologically similar earthworm species L. rubellus and E. andrei. The work indicates that 1H NMR spectroscopic methods provide a rapid means of profiling invertebrate biochemistry and may be of value in studies on the comparative toxicology of invertebrate species.