Sensitive fluorometric method for tissue tocopherol analysis

A sensitive, highly reproducible method for tissue tocopherol analysis that combines saponification in the presence of large nmount of ascorbic acid to remove interfering substances, extraction fo the nonsponifiable lipids with hexane, and fluorometric measurement of the tocopherol is presented. The nonsaponifiable lipids phase contained only one fluorochrome in the 290 am excitation and 330 nm emission range, and it was identified as tocopherol by thin layer and column chromatography. Column chromatography of the hexane extract of a saponified, 14C-tocopherolspiked microsomal fraction showed that no measurable oxidation to tocopherylquinone had occurred. The flurometric method for tocopherol analysis was applied to homogenates and subcellular fractions from rat liver, kidney, lung, and heart and red blood cells. The heavy mitochondrial and microsomal fractions had the highest subcellular concentrations of tocopherol.