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Separation of α-tocotrienol oxidation products and eight tocochromanols by HPLC with DAD and fluorescence detection and identification of unknown peaks by DAD, PBI-EIMS, FTIR, and NMR

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Abstract

Tocotrienols, like tocopherols, are members of the vitamin E family. While tocopherols (T) have been studied intensively, only recently have tocotrienols (T3) received increased attention due to their special health benefits. However, these positive attributes of T3 are probably lost as a result of degradation during food storage and processing, and there is little information about their oxidation products. Of particular interest are the oxidation products of α-tocotrienol (α-T3) as this is the least thermostable T3 isomer with the highest rate of degradation. The objective of this study was therefore to develop a reliable method for the determination of the most important oxidation products of α-T3 along with other tocochromanol isomers. We developed a high-performance liquid chromatography method with diode array detection, fluorescence detection, and a particle beam interface electron impact mass spectroscopy in order to separate the most important oxidation products of α-T3 (α-T3 spirodimers/spirotrimers, α-tocotrienoldihydroxy dimer, 7-formyl-β-tocotrienol (7-FβT3), 5-formyl-γ-tocotrienol (5-FγT3), α-tocotrienolquinone (α-T3Q), and α-T3Q dimers and α-tocotrienolquinone epoxides (α-T3QE)) from eight tocochromanol isomers. Furthermore, we sought to identify the as yet unknown oxidation products 5-FγT3, 7-FβT3, α-T3Q-dimer, and α-T3QE. Of these, 5-FγT3 was fully characterized by Fourier transform infrared spectroscopy and 1H and 13C nuclear magnetic resonance spectroscopy.

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Abbreviations

5-FγT:

5-Formyl-γ-tocopherol

5-FγT3:

5-Formyl-γ-tocotrienol

7-FβT:

7-Formyl-β-tocopherol

7-FβT3:

7-Formyl-β-tocotrienol

AIBN:

Azobisisobutyronitrile

DAD:

Diode array detection

EIMS:

Electron impact mass spectroscopy

F:

Fluorescence

FTIR:

Fourier transform infrared spectroscopy

HPLC:

High-performance liquid chromatography

MS:

Mass spectroscopy

NMR:

Nuclear magnetic resonance spectroscopy

PBI:

Particle beam interface

SIBL:

Strain-induced bond localization

TBME:

tert-Butyl methyl ether

UV–Vis:

Ultraviolet–visible spectroscopy

α-, β-, γ-, δ-T:

α- β-, γ-, δ-Tocopherol

α-, β-, γ-, δ-T3:

α-, β-, γ-, δ-Tocotrienol

α-T3-DHD:

α-Tocotrienol dihydroxydimer

α-T3Q:

α-Tocotrienolquinone

α-T3QE:

α-Tocotrienolquinone epoxide

α-T3QM:

α-Tocotrienolquinone methide

α-T3-SPD:

α-Tocotrienol spirodimer

α-T-DHD:

α-Tocopherol dihydroxydimer

α-TQ:

α-Tocopherolquinone

α-TQE:

α-Tocopherolquinone epoxide

α-TQM:

α-Tocopherolquinone methide

α-T-SPD:

α-Tocopherol spirodimer

α-T-SPT:

α-Tocopherol spirotrimers

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Acknowledgments

This work is part of the Food Network project funded by the Ministry for Science and Culture of Lower Saxony (Germany) via the Research Association of Agricultural and Nutritional Science of Lower Saxony (Forschungsverbund Agrar- und Ernährungswissenschaften Niedersachsen (FAEN)). The authors wish to gratefully acknowledge Leiber GmbH (Bramsche, Germany), Greenfox Produktions GmbH (Oldendorf/Luhe, Germany), and Davos Life Science (Singapore) for their support.

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  1. Department of Analytical Chemistry, Institute of Food Toxicology and Analytical Chemistry, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173, Hannover, Germany

    Anne Büsing & Waldemar Ternes

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Correspondence to Waldemar Ternes.

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Büsing, A., Ternes, W. Separation of α-tocotrienol oxidation products and eight tocochromanols by HPLC with DAD and fluorescence detection and identification of unknown peaks by DAD, PBI-EIMS, FTIR, and NMR. Anal Bioanal Chem 401, 2843–2854 (2011). https://doi.org/10.1007/s00216-011-5352-1

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