In vitro stable isotope labeling for discovery of novel metabolites by liquid chromatography-mass spectrometry: Confirmation of gamma-tocopherol metabolism in human A549 cell
Language: 
English
Short Title: 
In vitro stable isotope labeling for discovery of novel metabolites by liquid chromatography-mass spectrometry
Abstract: 

A general approach for discovering novel catabolic metabolites from a parent biocompound was developed and validated on the metabolism of gamma-tocopherol in human A549 cell. The method is based on LC-MS analysis of in vitro stable isotope-labeled metabolites and assumes that a parent compound and its metabolites share a common functional group that can be derivatized by well-documented reagents. In this method, two equal aliquots of extracted metabolites are separately derivatized with isotope-coded (heavy) and non-isotope-coded (light) form of derivatizing reagent, mixed at 1:1 ratio and analyzed using LC-MS. The metabolites with common functional group are then easily recognized by determination of a chromatographically co-eluted pair of isotopomers (MS doublet peaks) with similar peak intensities and mass difference corresponding to the mass difference between heavy and light form of derivatization reagent. The feasibility of this approach was demonstrated and validated by the identification of products of gamma-tocopherol catabolism in human A549 cell culture media using N-methyl-nicotinic acid N-hydroxysuccinimide ester (C1-NANHS) and N-methyl-d3-nicotinic acid N-hydroxysuccinimide ester (C1-d3-NANHS) derivatizing reagent. Overall four gamma-tocopherol metabolites were identified including 9'-COOH, 11'-COOH, 13'-COOH and 13'-OH. In addition, the developed LC-MS method can also be used for the fast and sensitive quantitative analysis of gamma-tocopherol and other forms of vitamin E related compounds.

Author(s): 
Yang, Wen-Chu
Regnier, Fred E.
Jiang, Qing
Adamec, Jiri
Item Type: 
Journal Article
Publication Title: 
Journal of Chromatography. A
Journal Abbreviation: 
J Chromatogr A
Publication Date: 
Jan 29, 2010
Publication Year: 
2010
Pages: 
667-675
Volume: 
1217
Issue: 
5
ISSN: 
1873-3778
DOI: 
10.1016/j.chroma.2009.12.002
Library Catalog: 
NCBI Published Medical (?)
Extra: 
PMID: 20006340 PMCID: PMC2813381

Turabian/Chicago Citation

Wen-Chu Yang, Fred E. Regnier, Qing Jiang and Jiri Adamec. Jan 29, 2010. "In vitro stable isotope labeling for discovery of novel metabolites by liquid chromatography-mass spectrometry: Confirmation of gamma-tocopherol metabolism in human A549 cell." Journal of Chromatography. A 1217: 5: 667-675. 10.1016/j.chroma.2009.12.002.

Wikipedia Citation

<ref> {{Cite journal | doi = 10.1016/j.chroma.2009.12.002 | issn = 1873-3778 | volume = 1217 | pages = 667-675 | last = Yang | first = Wen-Chu | coauthors = Regnier, Fred E., Jiang, Qing, Adamec, Jiri | title = In vitro stable isotope labeling for discovery of novel metabolites by liquid chromatography-mass spectrometry: Confirmation of gamma-tocopherol metabolism in human A549 cell | journal = Journal of Chromatography. A | date = Jan 29, 2010 | pmid = | pmc = }} </ref>