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

• Main Authors: Qing Liu, Jingwei Cai, Robert G. Nichols, Yuan Tian, Jintao Zhang, Philip B. Smith, Yan Wang, Chao Yan, Andrew D. Patterson
• Format: Article
• Language: English
• Published: MDPI AG 2019-06-01
• Series: Metabolites
• Subjects: polar metabolites; HILIC–UHPLC–MS/MS; targeted metabolomics; TCDD
• Online Access: https://www.mdpi.com/2218-1989/9/6/118

A hydrophilic interaction liquid chromatography (HILIC)&#8722;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-<i>p</i>-dioxin (TCDD). Comparisons of sensitivity (limit of detection as low as 0.01 &#181;M) 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&#8722;CoA and nucleosides such as UMP, XMP, and CMP were also markedly decreased. These results support the application of HILIC&#8722;UHPLC&#8722;MS/MS for robust and reliable analysis of the cellular response to environmentally relevant toxicants at lower doses.