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A Quantitative HILIC–MS/MS Assay of the Metabolic Response of Huh-7 Cells Exposed to 2,3,7,8-Tetrachlorodibenzo-p-Dioxin

1
School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China
2
Center for Molecular Toxicology and Carcinogenesis, Department of Veterinary and Biomedical Sciences, Pennsylvania State University, University Park, PA 16802, USA
3
CAS Key Laboratory of Magnetic Resonance in Biological Systems, National Centre for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences (CAS), Wuhan 430071, China
4
Huck Institute of the Life Sciences, University Park, PA 16802, USA
*
Author to whom correspondence should be addressed.
Metabolites 2019, 9(6), 118; https://doi.org/10.3390/metabo9060118
Received: 30 May 2019 / Revised: 11 June 2019 / Accepted: 13 June 2019 / Published: 20 June 2019
A hydrophilic interaction liquid chromatography (HILIC)–ultra high-pressure liquid chromatography (UHPLC) coupled with tandem mass spectrometry (MS/MS) method was developed and applied to profile metabolite changes in human Huh-7 cells exposed to the potent aryl hydrocarbon receptor (AHR) ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Comparisons of sensitivity (limit of detection as low as 0.01 µM) and reproducibility (84% of compounds had an interday relative standard deviation (RSD) less than 10.0%; 83% of compounds had an intraday RSD less than 15.0%) were assessed for all the metabolites. The exposure of Huh-7 cells to the hepatotoxic carcinogen TCDD at low doses (1 nM and 10 nM for 4 h and 24 h, respectively) was reflected by the disturbance of amino acid metabolism, energy metabolism (glycolysis, TCA cycle), and nucleic acid metabolism. TCDD caused a significant decrease in amino acids such as serine, alanine, and proline while promoting an increase in arginine levels with 24 h treatment. Energy metabolism intermediates such as phosphoenolpyruvate and acetyl–CoA and nucleosides such as UMP, XMP, and CMP were also markedly decreased. These results support the application of HILIC–UHPLC–MS/MS for robust and reliable analysis of the cellular response to environmentally relevant toxicants at lower doses. View Full-Text
Keywords: polar metabolites; HILIC–UHPLC–MS/MS; targeted metabolomics; TCDD polar metabolites; HILIC–UHPLC–MS/MS; targeted metabolomics; TCDD
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Liu, Q.; Cai, J.; Nichols, R.G.; Tian, Y.; Zhang, J.; Smith, P.B.; Wang, Y.; Yan, C.; Patterson, A.D. A Quantitative HILIC–MS/MS Assay of the Metabolic Response of Huh-7 Cells Exposed to 2,3,7,8-Tetrachlorodibenzo-p-Dioxin. Metabolites 2019, 9, 118.

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