J. Lipid Res. Neurobiology of Lipids (ISSN1683-5506)
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A more recent version of this article appeared on May 1, 2005

Papers In Press, published online ahead of print February 16, 2005
J. Lipid Res., doi:10.1194/jlr.D400044-JLR200
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Submitted on December 30, 2004
Revised on February 4, 2005
Accepted on February 6, 2005

Quantitation of rat liver vitamin E metabolites by LC-MS during high-dose vitamin E administration

Scott W. Leonard, Eric Gumpricht, Michael W. Devereaux, Ronald J. Sokol, and Maret G. Traber

Linus Pauling Institute, Oregon State University, Corvallis, Oregon 97331

Corresponding Author: scott.leonard{at}oregonstate.edu

To evaluate vitamin E metabolism, a method was developed to quantitate liver a- and g-tocopherol metabolites, a-CEHC (2,5,7,8-tetramethyl-2-(2'-carboxyethyl)-6-hydroxychroman) and g-CEHC (2,7,8-trimethyl-2-(2'-carboxyethyl)-6-hydroxychroman), respectively. Vitamin E supra-enriched livers were obtained from rats that were injected with vitamin E daily for 18 days. Liver samples (~50 mg) were homogenized, homogenate CEHC-conjugates hydrolyzed, CEHCs extracted with ethyl ether, then CEHCs were quantitated using liquid chromatography-mass spectrometery (LC-MS). Precision, based on inter-sample variability, ranged from 1-3%. Recovery of a- and g-CEHCs added to liver homogenates ranged from 77% to 87%. Detection limits of a- and g-CEHC were 20 fmol, with a linear detector response from 0.025 to 20 pmol injected. Corresponding with an increase in liver a-tocopherol (a-T), the MS peak for liver a-CEHC (m/z 277.8) increased 80-times (0.18 ± 0.01 to 15 ± 2 nmol/g). Liver a-CEHC concentrations were correlated with plasma a-CEHC, liver a-T, and plasma a-T (P < 0.001 for each comparison). a-CEHC represented 0.5 to 1% of the liver a-T concentration. Thus, LC-MS can be successfully used to quantitate a- and g-CEHC in liver samples. These data suggest that in times of excess liver a-T, increased metabolism of a-T to a-CEHC occurs.


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