TY - JOUR
T1 - An optimized procedure for metabonomic analysis of rat liver tissue using gas chromatography/time-of-flight mass spectrometry
AU - Pan, Li
AU - Qiu, Yunping
AU - Chen, Tianlu
AU - Lin, Jinchao
AU - Chi, Yi
AU - Su, Mingming
AU - Zhao, Aihua
AU - Jia, Wei
N1 - Funding Information:
This work is funded by the National Basic Research Program of China (Project numbers 2007CB914700, 2007CB511905), the Ministry of Science and Technology of the People's Republic of China (Project number 2006BAI08B04-01) and Research Projects of the Shanghai Science and Technology Commission (Project Number 064307067, 06DZ05906).
PY - 2010/8
Y1 - 2010/8
N2 - 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.
AB - 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.
KW - Carbon tetrachloride
KW - Gas chromatography/time-of-flight mass spectrometry
KW - Liver tissue
KW - Metabonomics
KW - Multivariate statistics
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U2 - 10.1016/j.jpba.2010.01.046
DO - 10.1016/j.jpba.2010.01.046
M3 - Article
C2 - 20185264
AN - SCOPUS:77950369068
SN - 0731-7085
VL - 52
SP - 589
EP - 596
JO - Journal of Pharmaceutical and Biomedical Analysis
JF - Journal of Pharmaceutical and Biomedical Analysis
IS - 4
ER -