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Comprehensive analysis of vitamin E constituents in human plasma by liquid chromatography-mass spectrometry

Comprehensive analysis of vitamin E constituents in human plasma by liquid chromatography-mass spectrometry

Analytical Chemistry 79(18): 7087-7096

The present paper describes the development and validation of a normal-phase liquid chromatography-mass spectrometry (NP-HPLC-MS) method for the screening and quantification of vitamin E constituents in human plasma and food matrixes. Liquid-liquid extraction combined with isotope dilution was applied to extract the lipophilic target analytes. Baseline separation of alpha-tocopherylacetate, alpha-tocopherol, alpha-tocotrienol, alpha-tocopherylquinone, beta-tocopherol, gamma-tocopherol, beta-tocotrienol, gamma-tocotrienol, delta-tocopherol, and delta-tocotrienol was achieved utilizing a normal-phase amine column operated with n-hexane and 1,4-dioxane as solvents. Detection was achieved by positive-ion atmospheric-pressure chemical ionization (APCI). Key features of the method are lower limits of detection, 3-51 nmoles/L; lower limits of quantification, 8-168 nmoles/L; linearity coefficients, 0.9778-0.9989; linear ranges, 0.01-29 micromol/L; recoveries, 53-92%; accuracies, 99-103%; intraday precisions, 2-17%; interday precisions, 5-18%; and suppression values, 0-29%. Fragmentation of tocopherols was studied by tandem mass spectrometry, and a fragmentation scheme for tocotrienols/tocopherols is postulated. Neutral-loss and precursor-ion scan experiments were performed for targeted discovery of oxidation products of tocopherols in human blood and fish oil, the latter being an important food component. The presented data suggest that this method will help to expand the number of quantified/discovered vitamin E constituents detected in food products and analyzed during human/animal trials in order to give a more comprehensive picture to nutritionists about the fate of vitamin E.

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Accession: 052269932

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PMID: 17696496

DOI: 10.1021/ac0708689

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