Comparison of 1-D and 2-D NMR techniques for screening earthworm responses to sub-lethal endosulfan exposure
Jimmy Yuk A , Jennifer R. McKelvie A , Myrna J. Simpson A , Manfred Spraul B and André J. Simpson A C
+ Author Affiliations
- Author Affiliations
A Department of Chemistry, University of Toronto, Scarborough College, 1265 Military Trail, Toronto, ON, M1C 1A4, Canada.
B Bruker BioSpin GmbH, Silberstreifen, D-76287 Rheinstetten, Germany.
C Corresponding author. Email: andre.simpson@utoronto.ca
Environmental Chemistry 7(6) 524-536 https://doi.org/10.1071/EN10084
Submitted: 28 July 2010 Accepted: 10 October 2010 Published: 21 December 2010
Environmental context. The application of metabolomics from an environmental perspective depends on the analytical ability to discriminate minute changes in the organism resulting from exposure. In this study, 1-D and 2-D Nuclear Magnetic Resonance (NMR) experiments were examined to characterise the earthworm’s metabolic response to an organochlorine pesticide. 2-D NMR showed considerable improvement in discriminating exposed worms from controls and in identifying the metabolites responsible. This study demonstrates the potential of 2-D NMR in understanding subtle biochemical responses resulting from environmental exposure.
Abstract. Nuclear Magnetic Resonance (NMR) based metabolomics is a powerful approach to monitoring an organism’s metabolic response to environmental exposure. However, the discrimination between exposed and control groups, depends largely on the NMR technique chosen. Here, three 1-D NMR and three 2-D NMR techniques were investigated for their ability to discriminate between control earthworms (Eisenia fetida) and those exposed to a sub-lethal concentration of a commonly occurring organochlorine pesticide, endosulfan. Partial least-squares discriminant analysis found 1H–13C Heteronuclear Single Quantum Coherence (HSQC) spectroscopy to have the highest discrimination with a MANOVA value (degree of separation) three orders lower than any of the 1-D and 2-D NMR techniques. HSQC spectroscopy identified alanine, leucine, lysine, glutamate, glucose and maltose as the major metabolites of exposure to endosulfan, more than all the other techniques combined. HSQC spectroscopy in combination with a shorter 1-D experiment may prove to be an effective tool for the discrimination and identification of significant metabolites in organisms under environmental stress.
Additional keywords: Eisenia fetida, metabolic profiling, metabolomics, Nuclear Magnetic Resonance.
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