PROTON NMR SPECTROSCOPIC STUDIES ON TISSUE-EXTRACTS OF INVERTEBRATE SPECIES WITH POLLUTION INDICATOR POTENTIAL

High resolution proton nuclear magnetic resonance (H-1 NMR) spectrosco pic methods have been used to characterise tissue extracts of a series of common British invertebrate species with pollution indicator poten tial. These include two earthworm species Lumbricus rubellus (Hoffmeis ter) and Eisenia andrei (Savigny), two terrestrial isopods, Oniscus as ellus (L.) and Porcellio scaber (Latreille), the diplopodous arthropod , Glomeris marginata (Villers) and a pulmonate gastropod, Arion subfus cus (Draparnaud). One and two-dimensional NMR techniques including H-1 -H-1 homonuclear correlation and H-1 J-Resolved NMR spectroscopic meth ods were applied to allow characterisation of the major organic compon ents in the tissue extracts. The extracts gave characteristic low mole cular weight metabolite NMR fingerprints for each species studied. End ogenous metabolites identified included glucose and trehalose, a range of free amino acids and organic acids and bases. The presence or abse nce of metabolites observed in the NMR spectra was examined by cluster analysis to investigate species similarity and differences in metabol ite profiles. The use of Principal Component Analysis to interrogate N MR data-reduced spectra of tissue extracts allowed for distinct separa tion of the two morphologically similar earthworm species L. rubellus and E. andrei. The work indicates that H-1 NMR spectroscopic methods p rovide a rapid means of profiling invertebrate biochemistry and may be of value in studies on the comparative toxicology of invertebrate spe cies. (C) 1997 Elsevier Science Inc.