TitleQuantitation of rat liver vitamin E metabolites by LC-MS during high-dose vitamin E administration.
Publication TypeJournal Article
Year of Publication2005
AuthorsLeonard SW, Gumpricht E, Devereaux MW, Sokol RJ, Traber MG
JournalJ Lipid Res
Date Published2005 May
KeywordsAnimals, Chromatography, Liquid, Kinetics, Male, Rats, Rats, Sprague-Dawley, Reproducibility of Results, Spectrometry, Mass, Electrospray Ionization, Tocopherols, Vitamin E

To evaluate vitamin E metabolism, a method was developed to quantitate liver alpha- and gamma-tocopherol metabolites, alpha-carboxyethyl hydroxychroman [alpha-CEHC; 2,5,7,8-tetramethyl-2-(2'-carboxyethyl)-6-hydroxychroman] and gamma-CEHC [2,7,8-trimethyl-2-(2'-carboxyethyl)-6-hydroxychroman], respectively. Vitamin E supraenriched livers were obtained from rats that were injected with vitamin E daily for 18 days. Liver samples (approximately 50 mg) were homogenized, homogenate CEHC-conjugates were hydrolyzed, CEHCs were extracted with ethyl ether, and then CEHCs were quantitated using liquid chromatography-mass spectrometry (LC-MS). Precision, based on intersample variability, ranged from 1% to 3%. Recovery of alpha- and gamma-CEHCs added to liver homogenates ranged from 77% to 87%. Detection limits of alpha- and gamma-CEHC were 20 fmol, with a linear detector response from 0.025 to 20 pmol injected. Corresponding with an increase in liver alpha-tocopherol, the MS peak for liver alpha-CEHC (mass-to-charge ratio 277.8) increased 80-fold (0.18 +/- 0.01 to 15 +/- 2 nmol/g). Liver alpha-CEHC concentrations were correlated with serum alpha-CEHC, liver alpha-tocopherol, and serum alpha-tocopherol (P < 0.001 for each comparison). alpha-CEHC represented 0.5-1% of the liver alpha-tocopherol concentration. Thus, LC-MS can be successfully used to quantitate alpha- and gamma-CEHC in liver samples. These data suggest that in times of excess liver alpha-tocopherol, increased metabolism of alpha-tocopherol to alpha-CEHC occurs.

Alternate JournalJ. Lipid Res.
PubMed ID15722564
Grant ListDK-38446 / DK / NIDDK NIH HHS / United States
DK-59576 / DK / NIDDK NIH HHS / United States
P30 ES-00210 / ES / NIEHS NIH HHS / United States