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Title: Development and validation of a gas chromatography/mass spectrometry method for the metabolic profiling of human colon tissue
Authors: Mal, M.
Koh, P.K.
Cheah, P.Y.
Chan, E.C.Y. 
Issue Date: 28-Feb-2009
Citation: Mal, M., Koh, P.K., Cheah, P.Y., Chan, E.C.Y. (2009-02-28). Development and validation of a gas chromatography/mass spectrometry method for the metabolic profiling of human colon tissue. Rapid Communications in Mass Spectrometry 23 (4) : 487-494. ScholarBank@NUS Repository.
Abstract: In this study, a gas chromatography/mass spectrometry (GC/MS) method was developed and validated for the metabolic profiling of human colon tissue. Each colon tissue sample (20 mg) was ultra-sonicated with 1mL of a mixture of chloroform/methanol/water in the ratio of 20:50:20 (v/v/v), followed by centrifugation, collection of supernatant, drying, removal of moisture using anhydrous toluene and finally derivatization using N-methyl-N-trifluoroacetamide (MSTFA) with 1% trimethylchlorosilane (TMCS). A volume of 1μL of the derivatized mixture was injected into the GC/MS system. A total of 53 endogenous metabolites were separated and identified in the GC/MS chromatogram, all of which were selected to evaluate the sample stability and precision of the method. Of the identified endogenous metabolites 19 belonging to diverse chemical classes and covering a wide range of the GC retention times (Rt) were selected to investigate the quantitative linearity of the method. The developed GC/MS method demonstrated good reproducibility with intra- and inter-day precision within relative standard deviation (RSD) of ±15%. The metabolic profiles of the intact tissue were determined to be stable (100 ± 15%) for up to 90 days at -80°C. Satisfactory results were also obtained in the case of other stability-indicating studies such as freeze/thaw cycle stability, bench-top stability and autosampler stability. The developed method showed a good linear response for each of the 19 analytes tested (r 2 > 0.99). Our GC/MS metabolic profiling method was successfully applied to discriminate biopsied colorectal cancer (CRC) tissue from their matched normal tissue obtained from six CRC patients using orthogonal partial least-squares discriminant analysis [two latent variables, R 2Y = 0.977 and Q 2 (cumulative) = 0.877]. Copyright © 2009 John Wiley & Sons, Ltd.
Source Title: Rapid Communications in Mass Spectrometry
ISSN: 09514198
DOI: 10.1002/rcm.3898
Appears in Collections:Staff Publications

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