Metabolism of a sea lamprey pesticide by fish liver enzymes part A: identification and synthesis of TFM metabolites
The sea lamprey (Petromyzon marinus) is a destructive invasive species in the Great Lakes that contributed to the collapse of native fish populations in the mid-1900s. 3-Trifluoromethyl-4-nitrophenol (TFM) is a selective pesticide that has been applied to sea lamprey infested tributaries of the Great Lakes to kill larvae since the 1960s and has reduced the populations by as much as 90%. However, the metabolism of TFM by sea lamprey and non-target species is not fully illuminated. Elucidation of TFM metabolism is critical for understanding its mode of action and possible environmental impact. Here, we describe the screening, identification, synthesis and structural characterization of TFM metabolites in livers from sea lamprey and three non-target species that differ in their ability to survive TFM exposure. We identified glucuronidation, sulfation, N-acetylation, glutathione conjugation, and aromatic nitro group reduction as potential detoxification mechanisms. Seven metabolites were synthesized for use as markers of TFM metabolism in fish. Quantitative 1H NMR was used to assay synthesized metabolite stock solutions that were then used as standard material to develop a quantitative LC-MS/MS method for TFM metabolites.
KeywordsTFM Nitroreduction Pesticide Biomarker synthesis Quantitative analysis LC-MS
This research was supported by the Great Lakes Fishery Commission. The authors thank Dr. Daniel Holmes from Michigan State University NMR core facility for his technical support. The authors also thank Andrew Paul Buchinger and Professor Kim Scribner for the gifts of fresh blue gill liver and fresh lake sturgeon liver. A.D.J. was supported by the USDA National Institute of Food and Agriculture and Hatch project MICL-02143.
Compliance with ethical standards
Animal tissues used in analyses were collected by staff of USGS Lake Huron Biological Station, Millersburg, MI 49759, USA. Experimental protocols involving the handling of fishes were carried out in accordance with United States federal guidelines for care and use of animals and were approved by the American Fisheries Society through the “Use of Fishes in Research Committee, 2014”.
Conflict of interest
The authors declare that they have no conflict of interest.
- 1.Mullett K, Sullivan P. Sea lamprey control in the Great Lakes. http://www.glfc.org/pubs/slcp/annual_reports/ANNUAL_REPORT_2016.pdf. Great Lakes Fishery Commission; 2016.
- 2.[Available from: http://www.glfc.org/.
- 5.Boogaard MA, Bills TD, Johnson DA. Acute toxicity of TFM and a TFM/niclosamide mixture to selected species of fish, including lake sturgeon (Acipenser fulvescens) and mudpuppies (Necturus maculosus), in laboratory and field exposures. J Great Lakes Res. 2003;29(Supplement 1(0)):529–41.CrossRefGoogle Scholar
- 11.Johanning K, Hancock G, Escher B, Adekola A, Bernhard MJ, Cowan-Ellsberry C, et al. Assessment of metabolic stability using the rainbow trout (Oncorhynchus mykiss) liver S9 fraction. Curr Protoc Toxicol. 2012;Chapter 14:Unit 14 0 1–28.Google Scholar
- 13.Duesel BF, Godfrey W. Process for the preparation of n-acetyl-p-aminophenol (apap). Google Patents; 1967.Google Scholar
- 14.Ho K-L, Murphy MB, Wan Y, Fong BMW, Tam S, Giesy JP, et al. Synthesis and characterization of bromophenol glucuronide and sulfate conjugates for their direct LC-MS/MS quantification in human urine as potential exposure markers for polybrominated diphenyl ethers. Anal Chem. 2012;84(22):9881–8.CrossRefGoogle Scholar
- 17.Bussy U, Delaforge M, El-Bekkali C, Ferchaud-Roucher V, Krempf M, Tea I, et al. Acebutolol and alprenolol metabolism predictions: comparative study of electrochemical and cytochrome P450-catalyzed reactions using liquid chromatography coupled to high-resolution mass spectrometry. Anal Bioanal Chem. 2013;405(18):6077–85.CrossRefGoogle Scholar
- 22.Burkhardt GN. 89. The action of chlorosulphonic acid on phenol and p-cresol at low temperatures. J Chem Soc (Resumed). 1933;337.Google Scholar
- 24.Brune K, Renner B, Tiegs G. Acetaminophen/paracetamol: a history of errors, failures and false decisions. Eur J Pain. 2014:n/a-n/a.Google Scholar
- 30.Bussy U, Chung-Davidson Y-W, Buchinger TJ, Li K, Smith SA, Jones DA, et al. Metabolism of a sea lamprey pesticide by fish liver enzymes. Part B. Method development and application in quantification of TFM metabolites formed in vivo. https://doi.org/10.1007/s00216-017-0831-7
- 31.Ellis F. Paracetamol—a curriculum resource. Osborne C, Pack M, editors: Royal Society of Chemistry; 2002. 26 p.Google Scholar
- 37.Dunlop ES, McLaughlin R, Adams JV, Jones M, Birceanu O, Christie MR, et al. Rapid evolution meets invasive species control: the potential for pesticide resistance in sea lamprey. Can J Fish Aquat Sci. 2017;1–17.Google Scholar