An optimized procedure for metabonomic analysis of rat liver tissue using gas chromatography/time-of-flight mass spectrometry

Li Pan, Yunping Qiu, Tianlu Chen, Jinchao Lin, Yi Chi, Mingming Su, Aihua Zhao, Wei Jia

Research output: Contribution to journalArticle

57 Scopus citations

Abstract

In this paper, we present a tissue metabonomic method with an optimized extraction procedure followed by instrumental analysis with gas chromatography/time-of-flight mass spectrometry (GC/TOFMS) and spectral data analysis with multivariate statistics. Metabolite extractions were carried out using three solvents: chloroform, methanol, and water, with design of experiment (DOE) theory and multivariate statistical analysis. A two-step metabolite extraction procedure was optimized using a mixed solvent of chloroform-methanol-water (1:2:1, v/v/v) and then followed by methanol alone. This approach was subsequently validated using standard compounds and liver tissues. Calibration curves were obtained in the range of 0.50-125.0μg/mL for standards and 0.02-0.25 g/mL acceptable for liver tissue samples. For most of the metabolites investigated, relative standard deviations (RSD) were below 10% within a day (reproducibility) and below 15% within a week (stability). Rat liver tissues of carbon tetrachloride-induced acute liver injury models (n=. 10) and healthy control rats (n=. 10) were analyzed which demonstrated the applicability of the developed procedure for the tissue metabonomic study.

Original languageEnglish (US)
Pages (from-to)589-596
Number of pages8
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume52
Issue number4
DOIs
StatePublished - Aug 1 2010
Externally publishedYes

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Keywords

  • Carbon tetrachloride
  • Gas chromatography/time-of-flight mass spectrometry
  • Liver tissue
  • Metabonomics
  • Multivariate statistics

ASJC Scopus subject areas

  • Analytical Chemistry
  • Pharmaceutical Science
  • Drug Discovery
  • Spectroscopy
  • Clinical Biochemistry

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