Metabonomic assessment of toxicity of 4-fluoroaniline, 3,5-difluoroaniline and 2-fluoro-4-methylaniline to the earthworm Eisenia veneta (Rosa): identification of new endogenous biomarkers.
High-resolution 1H nuclear magnetic resonance (NMR) spectroscopy can be used to produce a biochemical fingerprint of low-molecular-weight metabolites from complex biological mixtures such as tissue extracts and biofluids. Changes in such fingerprint profiles can be used to characterize the effects of toxic insult in in vivo systems. The technique is nonselective and requires little sample preparation or derivatization. In the present study, earthworms (Eisenia veneta) were exposed to three different model xenobiotics by a standard filter paper contact test, and toxicant-induced biochemical changes were then investigated by characterizing the changes in endogenous metabolites visible in 600-MHz 1H NMR spectra of tissue extracts. The NMR spectral intensities were converted to discrete numerical values and tabulated in order to provide data matrices suitable for multivariate analysis. Principal component analysis showed that changes had occurred in the biochemical profiles relative to the undosed controls. The 2-fluoro-4-methylaniline-treated worms showed a decrease in a resonance from a compound identified as 2-hexyl-5-ethyl-3-furansulfonate using a combination of high-performance liquid chromatography (HPLC)-Fourier transform mass spectrometry (IonSpec, Lake Forest, CA, USA) and 1H and 13C NMR spectroscopy. An increase in inosine monophosphate was also observed. The 4-fluoroaniline-treated worms showed a decrease in maltose concentrations, and 3,5-difluoroaniline exerted the same effect as 2-fluoro-4-methylaniline but to a lesser extent. These changes could potentially be used as novel biomarkers of xenobiotic toxicity and could be used to determine the mechanism of action of other toxic chemicals.
Imperial College of Science, Technology and Medicine, Biological Chemistry, Faculty of Medicine, South Kensington, London, United Kingdom. jgbundy@ucdavis.edu
Jacob G
JG
Bundy
Eva M
EM
Lenz
Nigel J
NJ
Bailey
Claire L
CL
Gavaghan
Claus
C
Svendsen
David
D
Spurgeon
Peter K
PK
Hankard
Daniel
D
Osborn
Jason M
JM
Weeks
Sunia A
SA
Trauger
Paul
P
Speir
Ian
I
Sanders
John C
JC
Lindon
Jeremy K
JK
Nicholson
Huiru
H
Tang
D000814:Aniline Compounds / Q000633:toxicity*; D000818:Animals; D015415:Biomarkers / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D007291:Inosine Monophosphate / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy; D008320:Maltose / Q000032:analysis; D009835:Oligochaeta; D015262:Xenobiotics / Q000633:toxicity*
D000814:Aniline Compounds; D015415:Biomarkers; D015262:Xenobiotics; D007291:Inosine Monophosphate; D008320:Maltose