Sensitive fluorometric method for tissue tocopherol analysis
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A sensitive, highly reproducible method for tissue tocopherol analysis that combines saponification in the presence of large amounts of ascorbic acid to remove interfering substances, extraction of the nonsaponifiable lipids with hexane, and fluorometric measurement of the tocopherol is presented. The nonsaponifiable lipid phase contained only one fluorochrome in the 290 nm 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 has occurred. The fluorometric 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.
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- 1.Bunnell, R.H., Lipids 6:245 (1971).Google Scholar
- 2.Bieri, J.G., in “Lipid Chromatographic Analysis”, Vol. 2, Edited by G.V. Marinetti, Marcel Dekker, New York, NY, 1969 pp. 459–478.Google Scholar
- 3.Bunnell, R.H., in “The Vitamins”, Vol. VI, Edited by P. Gyorgy and W.N. Pearson, Academic Press, New York, NY, 1967, pp. 265–316.Google Scholar
- 12.Cavins, J.F., and G.E. Inglett, Cereal Chem. 51:605 (1975).Google Scholar
- 14.Emmerie, A., and C. Engel, Rec. Trav. Chim. Pays-Bas 57:1351 (1938).Google Scholar
- 19.Hansen, L.G., and W.J. Warwick, Am. J. Clin. Pathol. 46:133 (1966).Google Scholar
- 23.de Duve, C., B.C. Pressman, R. Gianetto, R. Wattiaux, and F. Appellmans, Biochem. J. 60:604 (1955).Google Scholar
- 27.Bolliger, H.R., in “Thin-layer Chromatography: A Laboratory Handbook” Edited by E. Stahl, Academic Press, Inc., New York, NY, 1965, p. 232.Google Scholar