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An Optimized Procedure for Metabonomic Analysis of Rat Liver Tissue Using Gas Chromatography/Time-of-Flight Mass Spectrometry

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
Wei Jia, Professor and Co-Director of the UNCG Center for Research Excellence in Bioactive Food Components (Creator)
Yunping Qiu, Post Doctoral Fellow, Center for Research Excellence in Bioactive Food Components (Creator)
Institution
The University of North Carolina at Greensboro (UNCG )
Web Site: http://library.uncg.edu/

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 0.50–125.0 µg/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.

Additional Information

Publication
Journal of Pharmaceutical and Biomedical Analysis, 52, 589–596
Language: English
Date: 2010
Keywords
metabonomics, liver tissue, gas chromatography/time-of-flight mass spectrometry, multivariate statistics, carbon tetrachloride