In nature tocotrienols (compounds of vitamin E) have only been found in the 2R,3′trans,7′trans (R, trans-trans) configuration whereas synthetic tocotrienols often represent a mixture of cis and trans side chain isomers. In this study tocotrienols were isolated from samples (egg yolk, palm oil, and ewe's milk) by extraction of the unsaponifiable matter and subsequent semi-preparative high-performance liquid chromatography (HPLC). A recently developed HPLC method [two series-connected columns ET 200/4 NUCLEODEX β-PM (200×4 mm ID, permethylated β-cyclodextrin phase; Macherey-Nagel, Duren, Germany), acetonitrile/water eluent (58 : 42 by vol.), fluorescence detection at an emission wavelength of 330 nm (excitation wavelength = 295 nm)] was used to separate the isolated tocotrienols into the four potential cis/trans side chain isomers. Commercially available, synthetic α-tocotrienol was orally administered to a laying hen. In the egg yolk all four α-tocotrienol geometrical isomers could be determined, thus demonstrating the bioavailability. Furthermore, no cis/trans isomers of γ-tocotrienol from hardened palm oil (melting point 46 °C) could be detected, i.e. the hydrogenation process during oil hardening evidently caused no isomerization of γ-tocotrienol. Finally, it is shown that ewe's milk does not contain a detectable amount of cis/transα-tocotrienol isomers, which might derive from biohydrogenation during microbial metabolism in the rumen.
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