Please use this identifier to cite or link to this item: https://doi.org/10.1002/rcm.3898
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. https://doi.org/10.1002/rcm.3898
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
URI: http://scholarbank.nus.edu.sg/handle/10635/105821
ISSN: 09514198
DOI: 10.1002/rcm.3898
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

52
checked on Mar 26, 2020

WEB OF SCIENCETM
Citations

49
checked on Mar 18, 2020

Page view(s)

91
checked on Mar 28, 2020

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.