Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 879, Issue 13-14, Pages 961-967Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2011.03.009
Keywords
Metabonomics; Liver; UPLC-Q-TOF/MS; Metabolic profiling; Type 2 diabetes
Funding
- National Natural Science Foundation of China [20675082]
- State Key Science & Technology Project for Infectious Diseases [2008ZX10002-019, 2008ZX10002-017]
- State Ministry of Science & Technology of China [2007CB914701]
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A protocol for the metabolic profiling of rat liver was developed based on ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) to explore metabolic state directly. Methanol/water (4:1, v:v) was selected as the optimal extraction solvent. The established method was validated with a linearity over the 10-5000 ng/mL for internal standards (IS) and got an average correlation coefficient of 0.9986. The intra-day and inter-day RSD for most endogenous compounds were below 15%. And the absolute recovery of IS was from 84.8% to 109.1%. Liver tissues from diabetic and control rats were enrolled in the subsequent study to show the usefulness of the method. A clear classification between the control and model animals was achieved, some significant metabolites were successfully filtered. These metabolites reflected the abnormal metabolism of diabetic rats. This initial application indicated that the method is suitable and reliable for liver tissue metabolic profiling. It is expected this protocol could also be extended to metabonomic studies of other tissues. (C) 2011 Elsevier B.V. All rights reserved.
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