Journal
JOURNAL OF LIPID RESEARCH
Volume 56, Issue 11, Pages 2217-2225Publisher
ELSEVIER
DOI: 10.1194/jlr.D061663
Keywords
tocopherol; tocotrienol; metabolism; liquid chromatography tandem mass spectrometry; sulfation
Categories
Funding
- National Institutes of Health [R01AT006882, R01ES023349, P30CA023168]
- Indiana Clinical and Translational Sciences Institute - National Institutes of Health, National Center for Advancing Translational Sciences, Clinical and Translational Sciences Award [UL1 TR001108]
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Tocopherols and tocotrienols are metabolized via hydroxylation and oxidation of their hydrophobic side chain to generate 13-hydroxychromanols (13-OHs) and various carboxychromanols, which can be further metabolized by conjugation including sulfation. Recent studies indicate that long-chain carboxychromanols, especially 13-carboxychromanol (13-COOH), appear to be more bioactive than tocopherols in anti-inflammatory and anticancer actions. To understand the potential contribution of metabolites to vitamin E-mediated effects, an accurate assay is needed to evaluate bioavailability of these metabolites. Here we describe an LC/MS/MS assay for quantifying vitamin E metabolites using negative polarity ESI. This assay includes a reliable sample extraction procedure with efficacy of 89% and interday/intraday variation of 3-11% for major metabolites. To ensure accurate quantification, short-chain, long-chain, and sulfated carboxychromanols are included as external/internal standards. Using this assay, we observed that sulfated carboxychromanols are the primary metabolites in the plasma of rodents fed with -tocopherol or -tocopherol. Although plasma levels of 13-COOHs and 13-OHs are low, high concentrations of these compounds are found in feces. Our study demonstrates an LC/MS/MS assay for quantitation of sulfated and unconjugated vitamin E metabolites, and this assay will be useful for evaluating the role of these metabolites in vivo.
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