Gas chromatography-mass spectrometry-based profiling of serum fatty acids in acetaminophen-induced liver injured rats

J Appl Toxicol. 2014 Feb;34(2):149-57. doi: 10.1002/jat.2844. Epub 2012 Dec 12.

Abstract

In this study, we have developed and validated a simple, accurate and sensitive gas chromatography-mass spectrometry (GC-MS) method for simultaneous quantification of 18 fatty acids in rat serum, including both non-esterified (NEFA) and esterified (EFA) fatty acids, and subsequent analysis of fatty acid metabolic profiles. This novel method was used to evaluate the serum levels of fatty acids from vehicle- and acetaminophen (APAP)-treated rats. Serum levels of 7 NEFAs and 14 EFAs were significantly higher in APAP-treated rats 24 h after APAP administration at 1500 mg kg⁻¹ when compared with vehicle-treated controls. Control and APAP-treated rats could be differentiated based on their metabolic profiles using two different chemometric analysis methods: principle component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). More importantly, we identified the following NEFAs as potential biomarkers of APAP-induced liver injury: oleic acid (C18:1n9), linoleic acid (C18:2n6), docosahexaenoic acid (C22:6n3) and arachidonic acid (C20:4n6). The serum concentrations of C18:1n9, C18:2n6 and C22:6n3 were all positively correlated (r > 0.8; Pearson's correlation analysis) with the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). These results suggest that a novel targeted metabolomics method based on the metabolic profiling of fatty acids analyzed by GC-MS provides exact serum concentrations of fatty acids as well as a prospective methodology to evaluate chemically induced hepatotoxicity.

Keywords: GC-MS; acetaminophen; fatty acids; liver injury; metabolic profile.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetaminophen / adverse effects*
  • Alanine Transaminase / blood
  • Animals
  • Aspartate Aminotransferases / blood
  • Biomarkers / blood
  • Chemical and Drug Induced Liver Injury / blood*
  • Discriminant Analysis
  • Fatty Acids / blood*
  • Fatty Acids, Nonesterified / blood
  • Gas Chromatography-Mass Spectrometry
  • Linear Models
  • Liver / drug effects
  • Liver / metabolism
  • Male
  • Metabolome / drug effects
  • Multivariate Analysis
  • Principal Component Analysis
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Biomarkers
  • Fatty Acids
  • Fatty Acids, Nonesterified
  • Acetaminophen
  • Aspartate Aminotransferases
  • Alanine Transaminase