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.
An Optimized Procedure for Metabonomic Analysis of Rat Liver Tissue Using Gas Chromatography/Time-of-Flight Mass Spectrometry
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Created on 6/25/2012
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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